Two new species of Anoplophora Hope, 1839 from China (Coleoptera, Cerambycidae, Lamiinae).

Two new species of starry longhorn beetles are described from China, i.e. Anoplophora iadina sp. nov. from Yunnan and A. zibroides sp. nov. from Hunan and Guangxi. Color plates are presented to illustrate their diagnostic characters.

Function of PHEX mutations p.Glu145* and p.Trp749Arg in families with X-linked hypophosphatemic rickets by the negative regulation mechanism on FGF23 promoter transcription.

X-linked hypophosphatemic rickets (XLH) is characterized by increased circulating fibroblast growth factor 23 (FGF23) concentration caused by PHEX (NM_000444.5) mutations. Renal tubular resorption of phosphate is impaired, resulting in rickets and impaired bone mineralization. By phenotypic-genetic linkage analysis, two PHEX pathogenic mutations were found in two XLH families: c.433 G > T, p.Glu145* in exon 4 and c.2245 T > C, p.Trp749Arg in exon 22. Immunofluorescence showed that the localization of p.Glu145* and p.Trp749Arg mutant and secretory PHEX (secPHEX) changed, with decreased expression. In a HEK293T cell model co-transfected with PHEX, secPHEX, and FGF23, wild-type PHEX, secPHEX, and FGF23 proteins were distributed in the cell membrane or endoplasmic reticulum, while the mutant was located in the nuclear membrane and cytoplasm. qPCR of p.Glu145* revealed decreased PHEX and secPHEX mRNA expression in cells, with no difference in mRNA expression of p.Trp749Arg. Both mutations decreased intracellular PHEX endopeptidase activity. Western blot analysis showed decrease in mutant and secPHEX protein expression and no FGF23 protein expression in single-transfected PHEX and secPHEX cells. In cells co-transfected with FGF23, PHEX and secPHEX mutation promoted FGF23 expression. Dual-luciferase reporter gene was used to detect the effect of PHEX on FGF23 promoter. The dual-luciferase reporter gene showed that after PHEX overexpression, the activity of mutant firefly luciferase was significantly higher than that of wild type. The regulatory mechanism between PHEX and FGF23 is still unclear, but we found that PHEX is a direct transcriptional inhibitor of FGF23 and affects the expression of FGF23. This study verified the pathogenicity of the two variants and revealed the possible regulatory mechanism between PHEX and FGF23.

Review of the genus Hechinoschema Thomson, 1857 (Coleoptera: Cerambycidae: Lamiinae: Monochamini), with description of a new genus and two new species.

A taxonomic review of the genus Hechinoschema is presented. A new genus, Propedicellus gen. nov., is described, and two new species Propedicellus guoliangi sp. nov. and Propedicellus qiului sp. nov. are described from China. A new combination Propedicellus vitalisi (Pic, 1925) comb. nov. is proposed. A key to species of Propedicellus is presented.

Screening and function discussion of a hereditary renal tubular acidosis family pathogenic gene.

Hereditary distal renal tubular acidosis (dRTA) is a rare disease of H+ excretion defect of α-intercalated cells in renal collecting duct, caused by decreased V-ATPase function due to mutations in the ATP6V1B1 or ATP6V0A4 genes. In the present study, a genetic family with 5 members of the complete dRTA phenotype were found with distal tubule H+ secretion disorder, hypokalemia, osteoporosis, and kidney stones. A variant NM_020632.2:c.1631C > T (p.Ser544Leu) in exon 16 on an ATP6V0A4 gene associated with dRTA was detected by next generation sequencing target region capture technique and verified by Sanger sequencing, which suggested that except for one of the patients who did not receive the test, the other four patients all carried the p.S544L heterozygote. In transfected HEK293T cells, cells carrying p.S544L-mut showed early weaker ATPase activity and a slower Phi recovery rate after rapid acidification. By immunofluorescence localization, it was observed that the expression level of p.S544L-mut on the cell membrane increased and the distribution was uneven. Co-immunoprecipitation showed the a4 subunit of ATP6V0A4/p.S544L-mut could not bind to the B1 subunit, which might affect the correct assembly of V-ATPase. The present study of dRTA family suggests that the p.S544L variant may be inherited in a dominant manner.

Seasonal dynamics and spatial distribution pattern of Parapoynx crisonalis (Lepidoptera: Crambidae) on water chestnuts.

Parapoynx crisonalis (Walker, 1859) (Lepidoptera: Crambidae) is a major pest of aquatic vegetables and aquatic landscape plants. It has been responsible for causing considerable economic damage to water chestnut (Trapa natans) plants. In the Changsha vicinity of China, P. crisonalis has five generations a year. Populations of P. crisonalis were relatively low in April and began to rapidly rise at the beginning of May. At the end of July and early August, the population dropped dramatically. A rebound occurred at the end of August and early September, which was referred to as the second population peak. From then, until early November, the P. crisonalis population steadily diminished in preparation for overwintering. The primary factors influencing the seasonal dynamics of P. crisonalis were the climatic conditions, especially the temperature, and secondarily precipitation. Between May and October, the P. crisonalis adults were evenly distributed in the pond. In May and June, the eggs of P. crisonalis were present in an aggregate distribution, due to the effects of environmental heterogeneity. In July and August, however, they were found to be in a uniform distribution.

Age-stage, two-sex life table of Parapoynx crisonalis (Lepidoptera: Pyralidae) at different temperatures.

Parapoynx crisonalis is an important pest of many aquatic vegetables including water chestnuts. Understanding the relationship between temperature variations and the population growth rates of P. crisonalis is essential to predicting its population dynamics in water chestnuts ponds. These relationships were examined in this study based on the age-stage, two-sex life table of P. crisonalis developed in the laboratory at 21, 24, 27, 30, 33 and 36°C. The results showed that the values of Sxj (age-stage-specific survival rate), fxj (age-stage-specific fecundity), lx (age specific survival rate) and mx (age-specific fecundity) increased as the temperature rose from 21 to 27°C, then decreased from 30 to 36°C. Temperature also had a significant effect on the net reproductive rate (R0), gross reproductive rate (GRR), intrinsic rate of increase (r) and finite rate of increase (λ). The value of these parameters were at low levels at 21, 33, and 36°C. Further, the r value decreased as the temperature rose from 24 to 30°C, while the GRR reached its highest level at 27°C. The results indicated that optimal growth and development of P. crisonalis occurred at temperatures between 24°C to 30°C when compared to the lowest temperature (21°C) and higher temperatures of 33°C and 36°C.

Lentivirus-mediated short hairpin RNA interference targeting TNF-alpha in macrophages inhibits particle-induced inflammation and osteolysis in vitro and in vivo.

BACKGROUND: Aseptic loosening is a significant impediment to joint implant longevity. Prosthetic wear particles are postulated to play a central role in the onset and progression of periprosthetic osteolysis, leading to aseptic loosening of the prosthesis. METHODS: We investigated the inhibitory effects of a lentivirus-mediated short hairpin RNA that targets the TNF-alpha gene on the particle-induced inflammatory and osteolytic changes via macrophages both in vitro and in vivo. An siRNA sequence targeting the mouse TNF-alpha gene from four candidates, transcribed in vitro, was screened and identified. A lentivirus vector expressing short hairpin RNA (shRNA) was then constructed in order to facilitate efficient expression of TNF-alpha-siRNA. Lentivirus-mediated shRNA was transduced into cells of the mouse macrophage line RAW 264.7. Ceramic and titanium particles were introduced 24 h after lentivirus transduction to stimulate cells. TNF-alpha expression, represented by both mRNA and protein levels, was quantified with real-time PCR and ELISA at all time intervals. Lentivirus-mediated shRNA suspension was locally administered into the murine calvarial model, followed by local injection of particles. A multi-slice spiral CT scan was used to evaluate the osteolysis of the calvaria by detecting the width of the cranial sutures. RESULTS: Macrophages developed pseudopods when co-cultured with particles. Lentivirus-mediated shRNA was shown to effectively inhibit the expression of TNF-alpha at both the mRNA and protein levels in RAW 264.7. The multi-slice spiral CT scan showed that the lentivirus-mediated shRNA significantly suppressed osteolysis of mouse calvaria. CONCLUSIONS: Our investigation highlighted the results that lentivirus-mediated shRNA targeting the TNF-alpha gene successfully inhibited particle-induced inflammatory and osteolytic changes both in vitro and in vivo. Therefore, lentivirus-mediated gene therapy may provide a novel therapeutic approach to aseptic joint loosening.

miR-542-3p inhibits the growth and invasion of colorectal cancer cells through targeted regulation of cortactin.

Colorectal cancer is one of the most common malignancies. Previous studies have reported that cortactin (CTTN) is often overexpressed in tumors and is associated with metastasis and poor prognosis of patients. The abnormal expression of microRNAs (miRNAs or miRs) is closely related to the development and progression of various types of cancer, including colorectal cancer. However, little is known about the miRNAs targeting cortactin. In the present study, prediction using biological software revealed that cortactin has binding sites for miR-542-3p. Transfection with miR-542-3p mimic demonstrated that miR­542-3p reduced the expression of cortactin in colorectal cancer cells. Dual luciferase reporter assays further demonstrated that miR-542-3p regulated cortactin in a targeted manner and that miR-542-3p expression was significantly downregulated in colorectal cancer cells. A cell proliferation assay and Transwell migration assay were undertaken: we noted that miR­542-3p inhibited the proliferation and invasion of colorectal cancer cells while promoting their apoptosis. By contrast, cortactin acted antagonistically. When co-transfected with miR-542-3p mimic and CTTN overexpression vector, the inhibitory effect of miR-542-3p was blocked. This indicates that miR-542-3p regulates CTTN in a targeted manner to modulate the growth and invasion of colorectal cancer cells. The present study thus provides new targets for the prevention and treatment of colorectal cancer.

Self-assembled metallogels formed from N,N',N''-tris(4-pyridyl)trimesic amide in aqueous solution induced by Fe(III)/Fe(II) ions.

In this work, we report self-assembled metallogels formed from a ligand of trimesic amide, N,N',N''-tris(4-pyridyl)trimesic amide (TPTA), induced by Fe(III)/Fe(II) ions. TPTA is difficult to dissolve in water even in the presence of some metal ions such as Cu(2+), Co(2+), Ni(2+), K(+), Na(+) and Mg(2+) under heating, and it exhibits no gelation ability. Interestingly, upon heating TPTA can be dissolved easily in aqueous solution containing Fe(3+)/Fe(2+), and subsequently self-assembled into metallogels after cooling. The metallogels could also be formed in aqueous solutions of mixed metal ions containing Fe(3+)/Fe(2+), indicating that the other metal ions do not affect the formation of Fe(III)-TPTA and Fe(II)-TPTA metallogels. The high selectivity of metallogel formation to Fe(3+)/Fe(2+) may be used for application in the test of Fe(3+)/Fe(2+). The metallogels obtained are characterized by scanning electron microscopy, Fourier transform infrared spectra, nuclear magnetic resonance spectra, rheological measurements and scanning tunneling microscopy. The results indicate that TPTA can self-assemble into fibrous aggregates in Fe(3+)/Fe(2+) aqueous solution through the metal-ligand interactions and intermolecular hydrogen bonding. This kind of metallogel also possesses good mechanical properties and thermoreversibility.

Anti-tumor effect of LTA combined with 5-FU on H22 tumor bearing mice.

OBJECTIVE: To study the effect of lipoteichoic acid (LTA) and 5-FU on the expression of caspase-3, EGFR, TGF-α proteins of tumor tissue of H22 cancer bearing mice and its anti-tumor mechanism. METHODS: A total of 40 SPF grade Kunming mice were selected to establish H22 liver cancer model, and then the mice were divided into 4 groups at random with ten mice in each group. Group A was given saline lavage treatment, Group B was treated with 5-FU by intraperitoneal injection, Group C was treated with LTA by lump body injection; Group D was treated with LTA by lump body injection and 5-FU by intraperitoneal injection. Two weeks after the treatment, the mice in each group were executed and the tumor tissue was stripping and weighted, and the tumor growth inhibition ratio was calculated. Then the tumor tissue was processed for conventional embedding, sectioned to observe the expression of caspase-3, EGFR, TGF-α by immunohistochemical staining method. RESULTS: The tumor inhibitory rate o f Group D was significantly higher than Groups B and C (P < 0.05); B, the tumor inhibitory rate o f Group B had no statistical difference compared with Group C (P > 0.05). The IDO values of TGF-α, EGFR proteins in Groups B, C, D mice tumor tissue were significantly lower than that in group A (P < 0.05); while IDO value of caspase-3 in Groups B, C, D group mice tumor tissue was significantly higher than that in Group A (P < 0.05). The IDO value of TGF-α, EGFR in Group D mice tumor tissue were significantly lower than that in Groups B and C; While IDO value of aspase-3 in Group D was significantly higher than that in Groups B and C (P < 0.05). CONCLUSIONS: LTA combined with 5-FU can effectively inhibit the tumorigenesis of H22 tumor bearing mice, increase the caspase-3 protein expression, inhibit TGF-α and EGFR protein expression, further promote tumor cell apoptosis and play a synergistic antitumor effect.

Lentivirus vectors construction of SiRNA targeting interference GPC3 gene and its biological effects on liver cancer cell lines Huh-7.

OBJECTIVES: To build GPC3 gene short hairpin interference RNA (shRNA) slow virus vector, observe expression of Huh-7 GPC3 gene in human liver cell line proliferation apoptosis and the effect of GPC3 gene influencing on liver cancer cell growth, and provide theoretical basis for gene therapy of liver cancer. METHODS: Hepatocellular carcinoma cell line Huh-7 was transfected by a RNA interference technique. GPC3 gene expression in a variety of liver cancer cell lines was detected by fluorescence quantitative PCR. Targeted GPC3 gene sequences of small interfering RNA (siRNA) PGC-shRNA-GPC3 were restructured. Stable expression cell lines of siRNA were screened and established with the help of liposomes (lipofectamine(TM2000)) as carrier transfection of human liver cell lines. In order to validate siRNA interference efficiency, GPC3 siRNA mRNA expression was detected after transfection by using RT-PCR and Western blot. The absorbance value of the cells of blank group, untransfection group and transfection group, the cell cycle and cell apoptosis were calculated, and effects of GPC3 gene on Huh-7 cell proliferation and apoptosis were observed. RESULTS: In the liver cancer cell lines Huh-7, GPC3 gene showed high expression. PGC-shRNA-GPC3 recombinant plasmid was constructed successfully via sequencing validation. Stable recombinant plasmid transfected into liver cancer cell lines Huh-7 can obviously inhibit GPC3 mRNA expression level. CONCLUSIONS: The targeted GPC3 siRNA can effectively inhibit the expression of GPC3.

Dietary supplementation of Pediococcus pentosaceus enhances innate immunity, physiological health and resistance to Vibrio anguillarum in orange-spotted grouper (Epinephelus coioides).

Groupers (Epinephelus spp.) are economically important fish species in Southeast Asian aquaculture. Vibriosis caused by Vibro spp. is one of the severe bacterial diseases that devastate the grouper aquaculture industry. Probiotics have been reported to show the potential to enhance fish immunity and to antagonize pathogens. In our previous study, a lactic acid bacterium Pediococcus pentosaceus strain 4012 (LAB4012), isolated from cobia intestine, protects cobia from photobacteriosis after a 2-week feeding. In this study, we examined the potential of LAB4012 to be a probiotic for the orange-spotted grouper through feeding, thus to guard against vibriosis. In vitro, LAB4012 culture supernatant with low pH suppressed the growth of Vibrio anguillarum, and lactic acid in the metabolite of LAB4012 appeared to be the major factor to the growth inhibition of V. anguillarum. In vivo, the challenge test showed that the cumulative mortality of the LAB4012-fed groupers was significantly lower than that of the control fish after V. anguillarum infection. Supplementation of LAB4012 in commercial diet not only enhanced the growth rate and erythrocyte numbers of the groupers, but also regulated the gene expression of the pro-/anti-inflammatory cytokines. One day post-infection of V. anguillarum, the leukocyte numbers in the peripheral blood and the phagocytic activity of the head-kidney phagocytes in the LAB4012-fed groupers were found significantly increased, when compared with those without LAB4012-feeding. These results suggested that LAB4012 can be a dietary probiotic for groupers in modulating the immunity and protecting the groupers from V. anguillarum infection.

Diet supplementation of Pediococcus pentosaceus in cobia (Rachycentron canadum) enhances growth rate, respiratory burst and resistance against photobacteriosis.

Cobia (Rachycentron canadum) is an economically important fish species for aquaculture in tropical and sub-tropical areas. Cobia aquaculture industry has severely damaged due to photobacteriosis caused by Photobacterium damselae subsp. piscicida (Pdp), especially in Taiwan. Antibiotics and vaccines have been applied to control Pdp infection, but the efficacy has been inconsistent. One species of lactic acid bacteria, Pediococcus pentosaceus strain 4012 (LAB 4012), was isolated from the intestine of adult cobia, and its culture supernatant can effectively inhibit Pdp growth in vitro. The acidic pH derived from metabolic acids in LAB culture supernatant was demonstrated to be an important factor for the suppression. After a 2-week feeding of LAB 4012, the growth rate of the fed cobia was 12% higher than that of the non-fed group, and the relative percentage of survival (RPS) of the fed cobia was found to be 74.4 in Pdp immersion challenge. In addition, the respiratory burst (RB) of peripheral blood leukocytes (PBL) in the LAB 4012-fed group was significantly higher than that of the non-fed group. Although feeding LAB 4012 did not improve specific antibody response in cobia after immunization with Pdp vaccine, it still significantly raised the survival rate by 22% over that of the non-fed group after Pdp immersion challenge. Judging by the quick induction of high protection against Pdp infection and promotion of growth in larvae, LAB 4012 was considered to be a viable probiotic for cobia aquaculture.

rna interference targeting p110ß reduces tumor necrosis factor-alpha production in cellular response to wear particles in vitro and osteolysis in vivo.

Aseptic joint loosening is a key factor that reduces the life span of arthroplasty. There are currently few effective treatments for joint loosening except surgical revision. We explored the inhibitory effects of p110ß-targeted small interfering RNA (siRNA) and lentivirus on particle-induced inflammatory cytokine expression in the murine macrophage cell line, RAW264.7. siRNA- and lentivirus-targeting p110ß were transfected and infected prior to particle stimulation, respectively. Ceramic and titanium particles of different sizes were prepared to stimulate macrophages. Fluorescence microscopy showed that the efficiency of siRNA transfection and lentivirus infection were 74.2 ± 4.2 and 92.3 ± 2.6 %, respectively. TNF-alpha mRNA in the particle stimulation plus RNA interference (RNAi) groups were significantly lower compared with the particle stimulation-only groups (P < 0.05). Similarly, protein levels of TNF-alpha in RNAi-treated groups were significantly decreased after transfection or infection (P < 0.05). It showed that Phosphor-AKT (Ser473) activation was significantly reduced by RNAi through western blot. As assessed by CT, micro-CT and histological analysis, particle implantation induced a significant osteolysis in mice calvaria, which was limited by p110ß lentivirus addition. These results suggested that p110ß subtype of PI3K, followed by activation of Ser473, may possibly participate in the regulation of macrophages activity by wear particles, ultimately resulting in the TNF-α secretion and osteolysis.

Inhibition of the PI3K/AKT pathway reduces tumor necrosis factor-alpha production in the cellular response to wear particles in vitro.

Joint replacement is the most effective treatment for end-stage osteoarticular disease. However, macrophage-mediated aseptic loosening of joint prosthesis severely hampers the clinical effects of joint replacement. Until now, the mechanism by which macrophages regulate the secretion of inflammatory cytokines after particle stimulation is not clear. It is well known that the PI3K/AKT pathway participates in multiple cellular processes, including cell growth, survival, and inflammation. However, whether the PI3K/AKT pathway participates in the proinflammatory response of macrophages after particle stimulation and secondary aseptic loosening is still unknown. In this study, ceramic and titanium particles of different sizes were prepared to stimulate macrophages. LY294002, a specific inhibitor of PI3K, was pretreated prior to particle stimulation. The expression of tumor necrosis factor-alpha (TNF-α) and all the subunits of PI3K and AKT were detected by real-time polymerase chain reaction, enzyme-linked immunosorbent assay, and Western blot. The result showed that LY294002 could suppress the RNA and protein expression of TNF-α in RAW264.7 cells after stimulation of different particles. The subunits of PI3K (p110ß and p85ß), followed by activation of phosphor-AKT (Ser473), participated in the regulation of activating macrophages by wear particles, ultimately resulting in the secretion of TNF-α.

The involvement of AQP1 in heart oedema induced by global myocardial ischemia.

Aquaporin-1 (AQP1) is a member of aquaporin family that was previously proven to be involved in myocardial dysfunction; however, the role of AQP1 in myocardial stunning is less clear. To determine the change of AQP1 expression level in the heart and its effect on oedema after global myocardial ischemia, 40 adult goats underwent cardiopulmonary bypass (CPB) with an aortic cross-clamp time of 2 h and total bypass time of 6, 12, 24, 48 and 72 h followed by subsequent reperfusion. AQP1 function of eight goats was inhibited by HgCl(2) during the 24 h on CPB. All groups were compared with eight sham bypass control goats. Myocardial water content was measured, and the APQ1 mRNA and protein levels were detected by RT-PCR and immunoblotting, respectively. The results showed that the degree of myocardial oedema increased significantly at 6, 12, 24 and 48 h of reperfusion after CPB as compared with the control and recovered at 72 h of subsequent reperfusion. Expression levels of AQP1 mRNA and protein began to increase at 12 h and peaked at 24 h of CPB following reperfusion. Furthermore, myocardial oedema was reduced in the HgCl(2) group compared with the time-matched CPB and control groups. These data suggested that AQP1 expression increases in CPB and AQP1 plays an important role in myocardial oedema during CPB.

Surface behavior, aggregation and phase separation of aqueous mixtures of dodecyl trimethylammonium bromide and sodium oligoarene sulfonates: the transition to polyelectrolyte/surfactant behavior.

The properties and phase diagrams of aqueous mixtures of dodecyltrimethylammonium bromide (C(12)TAB) with the sodium oligoarene sulphonates (POSn), POS2, POS3, POS4, and POS6 have been studied using surface tension and neutron reflectometry to study the surface, and neutron small angle scattering and fluorescence to study the bulk solution. The behavior of POS2 and POS3 is reasonably consistent with mixed micelles of C(12)TAB and POSn-(C(12)TA)(n). These systems exhibit a single critical micelle concentration (CMC) at which the surface tension reaches the usual plateau. This is contrary to a recent report which suggests that the onset of the surface tension plateau does not coincide with the CMC. In the POS3 system, the micelles conform to the core-shell model, are slightly ellipsoidal, and have aggregation numbers in the range 70-100. In addition, the dissociation constant for ionization of the micelles is significantly lower than for free C(12)TAB micelles, indicating binding of the POS3 ion to the micelles. Estimation of the CMCs of the POSn-(C(12)TA)(n) from n = 1-3 assuming ideal mixing of the two component surfactants and the observed values of the mixed CMC gives values that are consistent with the nearest related gemini surfactant. The POS4 and POS6 systems are different. They both phase separate slowly to form a dilute and a concentrated (dense) phase. Fluorescence of POS4 has been used to show that the onset of aggregation of surfactant (critical aggregation concentration, CAC) occurs at the onset of the surface tension plateau and that, at the slightly higher concentration of the phase separation, the concentration of POS4 and C(12)TAB in the dilute phase is at or below its concentration at the CAC, that is, this is a clear case of complex coacervation. The surface layer of the C(12)TA ion in the surface tension plateau region, studied directly by neutron reflectometry, was found to be higher than a simple monolayer (observed for POS2 and POS3) for both the POS4 and POS6 systems. In POS6 this evolved after a few hours to a structure consisting of a monolayer with an attached subsurface bilayer, closely resembling that observed for one class of polyelectrolyte/surfactant mixtures. It is suggested that this structured layer, which must be present on the surface of the dilute phase of the coacervated system, is a thin wetting film of the dense phase. The close resemblance of the properties of the POS6 system to that of one large group of polyelectrolyte/surfactant mixtures shows that the surface behavior of oligoion/surfactant mixtures can quickly become representative of that of true polyelectrolyte/surfactant mixtures. In addition, the more precise characterization possible for the POS6 system identifies an unusual feature of the surface behavior of some polyelectrolyte/surfactant systems and that is that the surface tension can remain low and constant through a precipitation/coacervation region because of the characteristics of two phase wetting. The well-defined fixed charge distribution in POS6 also suggests that rigidity and charge separation are the factors that control whether a given system will exhibit a flat surface tension plateau or the alternative of a peak on the surface tension plateau.

Down-regulation of TNF-alpha by small interfering RNA inhibits particle-induced inflammation in vitro.

Aseptic joint loosening is a key factor that reduces the life span of arthroplasty. There are currently few effective treatments for joint loosening except surgical revision. We explored the inhibitory effects of tumor necrosis factor (TNF)-alpha-targeted small interfering RNA (siRNA) on particle-induced inflammatory cytokine expression in the murine macrophage cell line, RAW264.7. siRNA targeting TNF-alpha was chemically synthesized and transfected into RAW264.7 cells by cationic liposomes. Ceramic, titanium, and polyethylene particles of different sizes (0.2-1.2 µm and 1.2-10 µm) were prepared to stimulate cells 24 h after siRNA transfection. The down-regulation of TNF-alpha mRNA and protein levels was quantitatively determined using real-time polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA), respectively, at different time intervals. Fluorescence microscopy showed that the siRNA transfection efficiency was 85.2% ± 3.5%. Real-time PCR showed that the levels of TNF-alpha mRNA in the particle stimulation plus RNA interference (RNAi) groups were significantly lower compared with the particle stimulation-only groups 3 h after stimulation (P < 0.05). Similarly, ELISA essays showed that the protein levels of TNF-alpha in the RNAi-treated groups were significantly decreased 24 h after transfection (P < 0.05). There were no significant differences in the inhibitory effect among the groups stimulated by different particle types or particle sizes (P > 0.05). Our results indicated that siRNA targeting TNF-alpha can inhibit TNF-alpha release from RAW264.7 cells when stimulated by ceramic, titanium, and polyethylene particles of different sizes and that the inhibitory effects were not related to the type or size of particle.

Correlation between MR imaging and discography with provocative concordant pain in patients with low back pain.

OBJECTIVE: this study aimed to correlate magnetic resonance (MR) findings and discography with pain response at provocative discography in patients with low back pain. METHODS: ninety-three patients who underwent MR imaging of the lumbar spine and subsequent provocation discography as part of a clinical evaluation of low back pain were enrolled in the study. MR images were then evaluated for disc degeneration, high-intensity zone (HIZ), and endplate abnormalities. In the procedure of discography, concordant pain was denoted as positive, whereas discordant pain and no pain were denoted as negative. Finally, MR and discographic findings were analyzed by w2 test based on results of concordant pain. RESULTS: discography was conducted on 256 discs successfully, 116 discs of which presented with concordant pain, and the others presented with discordant pain. There were 141 discs we reevaluated as Grade I-III on MR images, 17 of which presented with concordant pain; 115 were evaluated as Grade IV-V, 99 of which presented with concordant pain. HIZ was found in 60 discs,52 of which had concordant pain. The endplate abnormalities we reobserved in 58 discs, 51 of which manifested concordant pain. Concordant pain was significant correlated with Type IV-V discs on discography (w2=144.08, r=0.60, P<0.01), Grade IV-V disc degeneration on MR image (w2=137.11, r=0.59, P<0.01), the presence of HIZ (w2=51.93, r=0.41, P<0.01), and endplate abnormalities (w2=52.76, r=0.41, P<0.01). DISCUSSION: disc degeneration grades on MR imaging showedan association with discographic grades. Type IV-V discs on discography, Grade IV-V disc on MR images, the presence of HIZ,and endplate abnormalities might indicate discogenic pain inpatients with chronic low back pain.

[Relationship between apnea and hyperbilirubinemia in premature infants].

OBJECTIVE: To explore the association between hyperbilirubinemia and apnea in premature infants. METHODS: Premature infants with apnea and birth weight >1500 g were tested for the heart rate, serum level of bilirubin, saturation of blood oxygen (SO2) and partial pressure of oxygen (PO2) before and after treatment, with term infants serving as the control. A comparative analyses of the serum level of bilirubin, SO2 and PO2 were carried out in the premature infants with birth weight <1500 g suffering apneic syndrome or not on the first and third days after birth. RESULTS: Of the premature and term infants with apnea and birth weight <1500 g, 92.5% and 70.00% showed increased serum level of indirect bilirubin (IBIL), respectively. The infants with birth weight <1500 g who presented the syndrome of apnea on the first day after birth had significantly higher levels of IBIL than those without an apparent syndrome of apnea. A three-day conventional therapy resulted in an obvious improvement of apneic syndrome and lowered bilirubin level. CONCLUSION: Increased bilirubin level can be one of the reasons for the development of apnea in premature infants, and therapies for reducing bilirubin level can ameliorate the syndrome of apnea.