[An investigation of musculoskeletal disorders at multiple sites and related influencing factors among workers in an automobile assembly shop].

Objective: To investigate the current status of work-related musculoskeletal disorders (WMSDs) in automobile assembly workers, as well as the distribution of WMSDs at multiple sites and related influencing factors. Methods: From March to July 2019, cluster sampling was performed to select 663 male automobile assembly workers as respondents, and the musculoskeletal injury questionnaire was used to investigate their general status and working condition. A multinomial logistic regression analysis was used to analyze the influencing factors for WMSDs at multiple sites. Results: The detection rate of WMSDs within the past 7 days was 37.9% (251/663) among the automobile assembly workers, and the detection rate of WMSDs within the past 1 year was 51.9% (344/663) . Of all workers, 13.6% (90/663) had WMSDs involving only 1 site, while 38.3% (254/663) had WMSDs involving 2 or more sites. The multinomial logistic regression analysis showed that frequent turns during work was a risk factor for WMSDs involving 1-3, 4-6, and 7-9 sites (odds ratio [OR]=1.65, 2.47, and 3.65, respectively) . Repeated action of lower extremities and ankles was a risk factor for WMSDs involving 4-6 and 7-9 sites (OR=2.15 and 2.98, respectively) . Working in an uncomfortable position was a risk factor for WMSDs involving 1-3, 4-6, and 7-9 sites (OR=1.95, 2.67, and 3.04, respectively) . Prolonged standing during work was a risk factor for WMSDs involving 1-3 and 4-6 sites (OR= 1.87 and 1.79, respectively) . Working overtime was a risk factor for WMSDs involving 7-9 sites (OR=5.48) . Adequate time for rest was a protective factor against WMSDs involving 1-3 and 4-6 sites (OR=0.50 and 0.31, respectively) . Conclusion: There is a high detection rate of WMSDs in automobile assembly workers, and WMSDs at multiple sites are more common than WMSDs at a single site. Poor position and organizational management factors are risk factors for occupational WMSDs at multiple sites.

[Study of local application of CpG-oligodeoxynucleotide combined with 4-1BB monoclonal antibody to treat hepatoma- bearing mice].

Objective: To investigate the curative effect of local application of CpG-oligodeoxynucleotide (CpG-ODN) combined with 4-1BB monoclonal antibody in hepatoma-bearing mice, and to evaluate the effect of 4-1BB monoclonal antibody on CpG-ODN immunotherapy. Methods: H22 single cell suspension was injected subcutaneously into the axilla and four limbs of the BALB/c male mice to establish a tumor-bearing mice model. After 7 days, 30 mice with corresponding tumor-bearing volume were screened and randomly divided into model control group, CpG group and CpG+4-1BB group, and the drug was injected into the tumors of left lower extremity. The same batch of normal mice was selected as normal control group. Survival of mice was recorded. Tumor-bearing volume and organ index were calculated. Serum levels of interleukin (IL) - 12 and interferon (IFN) gamma and spleen CD8(+)T lymphocyte ratio were measured. The measurement data were analyzed by analysis of variance. The survival rate of each group of mice was analyzed by log-rank test. Results: Mice in the model control group with tumor-bearing volume had a sustained growth before the execution. CpG group and the CpG+4-1BB group [(976.08 ± 29.55) mm(3), (47.25 ± 0.93) mm(3))] tumor-bearing volume was decreased than model group [(1 336.52 ± 39.40) mm3] (F = 5 329.273, P < 0.05). CpG+4-1BB group distant tumor-bearing volume [(611.83 ± 113.02) mm3] was decreased than model group and CpG group [(1 406.62 ± 51.09) mm(3), (1 380.01 ± 51.44) mm3] (F = 247.160, P < 0.05), but there was no significant difference between the CpG group and the model group (P > 0.05). Serum IL-12 concentration (23.90 ± 2.33 pg/ml), IFN-γ concentration (103.02 ± 6.10 pg/ml) and spleen CD8(+)T cell ratio (4.54 ± 0.62%) in the model group were lower than those in the normal group (P < 0.05). Serum IL-12 concentration in CpG group and CpG+4-1BB group (29.21 ± 2.23 pg/ml, 37.04 ± 1.49 pg/ml), IFN-γ concentration (116.12 ± 4.08 pg/ml, 138.65 ± 1.72 pg/ml), CD8(+)T cell ratio (6.65 ± 0.64%, 12.73 ± 0.88%) were higher than the model group, while CpG+4-1BB group was higher than the CpG group (P < 0.05). The survival rate of CpG+4-1BB group was higher than that of model group and CpG group (χ(2) = 25.544, P < 0.05), but there was no significant difference between CpG group and model group (P > 0.05). There was no significant difference in organ index between the four groups (P > 0.05). Conclusion: 4-1BB monoclonal antibody combined with CpG-ODN therapy can shrink hepatoma-bearing capacity, inhibit the growth of distant tumors and significantly prolong the survival time of mice.

Xanthotoxin prevents bone loss in ovariectomized mice through the inhibition of RANKL-induced osteoclastogenesis.

UNLABELLED: Xanthotoxin (XAT) is extracted from the seeds of Ammi majus. Here, we reported that XAT has an inhibitory effect on osteoclastogenesis in vitro through the suppression of both receptor activator of nuclear factor-κB ligand (RANKL)-induced ROS generation and Ca(2+) oscillations. In vivo studies showed that XAT treatment decreases the osteoclast number, prevents bone loss, and restores bone strength in ovariectomized mice. INTRODUCTION: Excessive osteoclast formation and the resultant increase in bone resorption activity are key pathogenic factors of osteoporosis. In the present study, we have investigated the effects of XAT, a natural furanocoumarin, on the RANKL-mediated osteoclastogenesis in vitro and on ovariectomy-mediated bone loss in vivo. METHODS: Cytotoxicity of XAT was evaluated using bone marrow macrophages (BMMs). Osteoclast differentiation, formation, and fusion were assessed using the tartrate-resistant acid phosphatase (TRAP) stain, the actin cytoskeleton and focal adhesion (FAK) stain, and the fusion assay, respectively. Osteoclastic bone resorption was evaluated using the pit formation assay. Reactive oxygen species (ROS) generation and removal were evaluated using dichlorodihydrofluorescein diacetate (DCFH-DA). Ca(2+) oscillations and their downstream signaling targets were then detected. The ovariectomized (OVX) mouse model was adopted for our in vivo studies. RESULTS: In vitro assays revealed that XAT inhibited the differentiation, formation, fusion, and bone resorption activity of osteoclasts. The inhibitory effect of XAT on osteoclastogenesis was associated with decreased intracellular ROS generation. XAT treatment also suppressed RANKL-induced Ca(2+) oscillations and the activation of the resultant downstream calcium-CaMKK/PYK2 signaling. Through these two mechanisms, XAT downregulated the key osteoclastogenic factors nuclear factor of activated T cells c1 (NFATc1) and c-FOS. Our in vivo studies showed that XAT treatment decreases the osteoclast number, prevents bone loss, rescues bone microarchitecture, and restores bone strength in OVX mice. CONCLUSION: Our findings indicate that XAT is protective against ovariectomy-mediated bone loss through the inhibition of RANKL-mediated osteoclastogenesis. Therefore, XAT may be considered to be a new therapeutic candidate for treating osteoporosis.

[Establishment of a cell co-culture system in accordance with the immunological characteristics of chronic HBV infection].

Objective: To investigate whether the co-culture of Huh7.93 cells and peripheral blood mononucleated cells from chronic hepatitis B patients (cPBMCs) can simulate the replication features of hepatitis B virus (HBV) and immune function in chronic hepatitis B (CHB) patients, and to provide an in vitro cell co-culture system for the research on immune clearance in chronic HBV infection. Methods: Huh7.93 cells were cultured alone or co-cultured with peripheral blood mononucleated cells from healthy people who underwent physical examination (nPBMCs) or cPBMCs for 7 days. The CCK8 assay was performed to measure the proliferative activity of Huh7.93 cells, and quantitative real-time PCR and Southern blot were used to measure HBV replication in cPBMCs and co-cultured cells. The independent samples t-test was used for comparison between two groups. Results: When Huh7.93 cells were co-cultured with peripheral blood mononucleated cells (PBMCs) at a certain ratio, Huh7.93 cells had a high proliferative activity and good cell growth. HBV could not infect or replicate in cPBMCs. HBV DNA in the supernatant of Huh7.93 cells co-cultured with cPBMCs showed significant increases and significantly higher than that in the supernatant of Huh7.93 cells cultured alone on day 4 (6.01 ± 0.20 log10copies/ml vs 4.99 ± 0.08 log10copies/ml, P = 0.000) and day 7 (7.82 ± 0.24 log10copies/ml vs 6.96±0.09 log10copies/ml, P = 0.000). On day 7 of culture, the cell lysis buffer of Huh7.93 cells co-cultured with cPBMCs had a significant increase in the level of HBV replicative intermediate compared with that of Huh7.93 cells cultured alone. After HepG2.2.15 cells were co-cultured with cPBMCs, there was no significant increase in HBV replication. Conclusion: The co-culture of Huh7.93 cells and cPBMCs produces similar viral replication as human body infected with HBV and can well simulate the liver microenvironment and immune function in CHB patients, which provides a new method for the research on immune clearance in chronic HBV infection.

[Application of three-dimensional visualized model of impacted tooth for surgical extraction in undergraduate oral experimental teaching].

Objective: To evaluate the effect of the application of a self-developed three-dimensional visualized model of impacted tooth for surgical extraction in undergraduate oral experimental teaching. Methods: Forty-one undergraduates majoring stomatology of Tongji University from 2018 [19 males and 22 females, aged (22.4±0.8) years] were enrolled and randomly divided into the conventional group and the experimental group. Students of the conventional group (21 students including 8 males and 13 females) received a teaching protocol for the surgical extraction of impacted tooth including theoretical lectures, watching operation videos, and operating on head-simulator teaching systems. Students of the experimental group (20 students including 11 males and 9 females) received an additional training of resistance assessment and surgical extraction using the three-dimensional visualized model of impacted tooth before operating on the head simulators. After class, a questionnaire survey was carried out among students, and the operation results on the head simulators were evaluated by the teacher. Results: The results of the questionnaire showed that the students in the experimental group were rated higher than those in the traditional group in terms of being able to imagine the relationship between impacted teeth and adjacent structures (U=114.00, P=0.006), avoiding damage to adjacent teeth (U=87.00, P<0.001) and inferior alveolar nerve during tooth extraction (U=111.50, P=0.006), and being more confident in clinical operations in the future (U=120.00, P=0.013). According to the evaluation results of tooth extraction on the head simulators, there was no significant difference in the operation time between the two groups (U=138.50, P=0.056). In the experimental group, 5% (1/20) caused adjacent tooth loosening and 15% (3/20) caused the excessive bone defect, which was less than those in the traditional group [38% (8/21) and 48% (10/21), respectively] (P=0.021; P=0.043). There was no significant difference in the incidence of grinding out the deep bone of impacted teeth between the two groups (P=0.232). Conclusions: The application of three-dimensional visualized model of impacted tooth for surgical extraction in undergraduate oral experimental teaching had achieved good results and was worth popularizing.

Structural characterization of cassava linamarase-linamarin enzyme complex: an integrated computational approach.

Cassava linamarase is a hydrolyzing enzyme that belongs to a glycoside hydrolase family 1 (GH1). It is responsible for breaking down linamarin to toxic cyanide. The enzyme provides a defensive mechanism for plants against herbivores and has various applications in many fields. Understanding the structure of linamarase at the molecular level is a key to avail its reaction mechanism. In this study, the three-dimensional (3D) structure of linamarase was built for the first time using homology modelling and used to study its interaction with linamarin. Molecular docking calculations established the binding and orientation nature of linamarin, while molecular dynamics (MD) simulation established protein-ligand complexes' stability. Binding-free energy based on MM/PBSA was further used to rescore the docking results. An ensemble structure was found to be relatively stable compared to the modelled structure. This study sheds light on the exploration of linamarase towards understanding its reaction mechanisms.Communicated by Ramaswamy H. Sarma.

Structure-based virtual screening and molecular dynamics simulation studies to discover new SARS-CoV-2 main protease inhibitors.

Computational methods were used to filter two datasets (> 8,000 compounds) based on two criteria: higher binding affinity for MPRO than cocrystallized inhibitor and binding interactions with MPRO catalytic dyad (Cys145 and His41). After virtual screening involving ranking and reranking, eleven compounds were identified to satisfy these criteria and analysis of their structures revealed an unparallel common features among them which could be critical for their interactions with MPRO. However, only the topmost scoring compound (AV-203: K i = 0.31 µM) exhibited relatively stable binding interaction during the period of 50 ns MD simulation and thus is a suitable template for drug development.

Determining the risk relationship associated with inferior alveolar nerve injury following removal of mandibular third molar teeth: A systematic review.

PURPOSE: This study analyzes the risk factors associated with the incidences of inferior alveolar nerve (IAN) injury after surgical removal of impacted mandibular third molar (IMTM) and to evaluate the contribution of these risk factors to postoperative neurosensory deficits. MATERIALS AND METHODS: An exhaustive literature search has been carried out in the COCHRANE library and PubMed electronic databases from January 1990 to March 2019 supplemented by manual searching to identify the related studies. Twenty-three studies out of 693 articles from the initial search were finally included, which summed up a total of 26,427 patients (44,171 teeth). RESULTS: Our results have been compared with other current available papers in the literature review that obtained similar outcomes. Among 44,171 IMTM extractions performed by various grades of operators, 1.20% developed transient IAN deficit and 0.28% developed permanent IAN deficit respectively. Depth of impaction (P<0.001), contact between mandibular canal (MC) and IMTM (P<0.001), surgical technique (P<0.001), intra-operative nerve exposure (P<0.001), and surgeon's experience (P<0.001) were statistically significant as contributing risk factors of IAN deficits. CONCLUSION: Radiographic findings, such as depth of impaction, proximity of the tooth to the mandibular canal, surgical technique, intra-operative nerve exposure, and surgeon's experience were high risk factors of IAN deficit after surgical removal of IMTMs.

HIF-1α Mediates Osteoclast-Induced Mandibular Condyle Growth via AMPK Signaling.

During the mandibular condylar growth, the absorption of calcified cartilage matrix induced by osteoclasts is crucial for the continuous endochondral osteogenesis. Meanwhile, recent studies showed that subchondral bone resided within the low-oxygen microenvironment, and our previous study revealed that hypoxia-inducible transcription factor 1α (HIF-1α) promoted osteoclastogenesis under hypoxia. However, whether HIF-1α regulates the function of osteoclasts in the mandibular condyle cartilage remains elusive. Our study indicated that severe deformity of the mandibular condyle was displayed in 10-wk-old osteoclast-specific HIF-1α conditional knockout (CKO) mice, accompanied by shortened length of condylar process and disorganized fibrocartilage. In 1-, 2-, and 4-wk-old CKO mice, the size of the hypertrophic layer and chondrocytic layer was significantly thickened. In the chondrocytic layer, chondrocytes were atrophied, showing a form of apoptosis in 4-wk-old CKO mice. Furthermore, an increase in the thickness of the fibrous and proliferating layer was observed in 10-wk-old CKO mice, as well as a significant decrease in that of the chondrocytic and hypertrophic chondrocyte layers. Interestingly, the articular surface of the condylar process abnormally presented a horizontal concave shape, and a disk-like acellular connective tissue appeared. In addition, genetic ablation of HIF-1α blunted cartilage matrix loss by subchondral osteoclast deficiency, resulting in a high subchondral bone mass phenotype, accompanied with a decreased number of blood vessels, alkaline phosphatase staining, and vascular endothelial growth factor (VEGF) expression. Mechanistically, the number of osteoclasts in the center of the condyle in CKO mice was significantly reduced by attenuated expression of adenosine 5'-monophosphate-activated protein kinase (AMPK) signaling. These findings reveal a novel influence of HIF-1α function in osteoclasts on maintenance of osteoclast-induced resorption of calcified cartilage matrix via AMPK signaling, as well as subchondral bone formation through VEGF-dependent angiogenesis in bone marrow.

Vinculin proteolysis unmasks an ActA homolog for actin-based Shigella motility.

To generate the forces needed for motility, the plasma membranes of nonmuscle cells adopt an activated state that dynamically reorganizes the actin cytoskeleton. By usurping components from focal contacts and the actin cytoskeleton, the intracellular pathogens Shigella flexneri and Listeria monocytogenes use molecular mimicry to create their own actin-based motors. We raised an antibody (designated FS-1) against the FEFPPPPTDE sequence of Listeria ActA, and this antibody: (a) localized at the trailing end of motile intracellular Shigella, (b) inhibited intracellular locomotion upon microinjection of Shigella-infected cells, and (c) cross-reacted with the proteolytically derived 90-kD human vinculin head fragment that contains the Vinc-1 oligoproline sequence, PDFPPPPPDL. Antibody FS-1 reacted only weakly with full-length vinculin, suggesting that the Vinc-1 sequence in full-length vinculin may be masked by its tail region and that this sequence is unmasked by proteolysis. Immunofluoresence staining with a monoclonal antibody against the head region of vinculin (Vin 11-5) localized to the back of motile bacteria (an identical staining pattern observed with the anti-ActA FS-1 antibody), indicating that motile bacteria attract a form of vinculin containing an unmasked Vinc-1 oligoproline sequence. Microinjection of submicromolar concentrations of a synthetic Vinc-1 peptide arrested Shigella intracellular motility, underscoring the functional importance of this sequence. Western blots revealed that Shigella infection induces vinculin proteolysis in PtK2 cells and generates p90 head fragment over the same 1-3 h time frame when intracellular bacteria move within the host cell cytoplasm. We also discovered that microinjected p90, but not full-length vinculin, accelerates rates of pathogen motility by a factor of 3 +/- 0.4 in Shigella-infected PtK2 cells. These experiments suggest that vinculin p90 is a rate-limiting component in actin-based Shigella motility, and that supplementing cells with p90 stimulates rocket tail growth. Earlier findings demonstrated that vinculin p90 binds to IcsA (Suzuki, T.A., S. Saga, and C. Sasakawa. 1996. J. Biol. Chem. 271:21878-21885) and to vasodilator-stimulated phosphoprotein (VASP) (Brindle, N.P.J., M. R. Hold, J.E. Davies, C.J. Price, and D.R. Critchley. 1996. Biochem. J. 318:753-757). We now offer a working model in which proteolysis unmasks vinculin's ActA-like oligoproline sequence. Unmasking of this site serves as a molecular switch that initiates assembly of an actin-based motility complex containing VASP and profilin.

Male requires a higher median target effect-site concentration of propofol for I-gel placement when combined with dexmedetomidine.

OBJECTIVE: The supraglottic airway device (SAD) can be used for airway management of spontaneous breathing patients, and propofol is commonly applied for the SAD placement. This study was designed to assess the effect of gender on median target effect-site concentration (Ce50) of propofol for I-gel placement when combined with dexmedetomidine. MATERIAL AND METHOD: 19 males and 18 females, aged 18 to 59 and undergoing elective surgery, were enrolled. After intravenous infusion of dexmedetomidine 1.0µg/kg over 10min followed by continuous infusion of 0.4µg/kg/h, target-controlled infusion of propofol under Marsh model was started and the initial Ce of propofol was set at 4.79µg/mL and 4.35µg/mL in the male and female patients, respectively. The I-gel was inserted when the Ce of propofol reached the pre-set concentration and bispectral index value was less than 60. The Ce of propofol required for I-gel placement was determined by the Dixon up-and-down method. RESULTS: The Ce50 (95% confidence interval) of propofol required for I-gel placement were 4.082µg/mL (3.798-4.332µg/mL) and 3.509µg/mL (3.266-3.749µg/mL) in male and female patients, respectively, with a significantly higher Ce50 in males. CONCLUSION: When combined with dexmedetomidine, males require a higher Ce50 of propofol for I-gel placement compared to females.

Circular RNA circRNA_0000285 promotes cervical cancer development by regulating FUS.

OBJECTIVE: Recently, the vital role of circular RNAs (circRNAs) in human diseases has attracted much attention. The aim of this research was to verify the potential role of circRNA_0000285 in the development of cervical cancer (CC). PATIENTS AND METHODS: CircRNA_0000285 expression in both CC cells and tissue samples was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Functional experiments were performed, including cell counting kit-8 (CCK-8) assay, cell cycle assay and transwell assay. Meanwhile, the underlying mechanism was explored through qRT-PCR and Western blot assay, respectively. In addition, the function of circRNA_0000285 was identified in vivo. RESULTS: CircRNA_0000285 expression level was significantly higher in CC samples than that of corresponding normal tissues. Moreover, the growth and migration abilities of CC cells were significantly inhibited after circRNA_0000285 was knocked down in vitro. Furthermore, the expression of FUS was remarkably down-regulated after knockdown of circRNA_0000285. Subsequent results indicated that the expression level of FUS was positively correlated with the expression of circRNA_0000285 in CC tissues. In addition, the knockdown of circRNA_0000285 significantly inhibited the formation and metastasis of CC in nude mice. CONCLUSIONS: CircRNA_0000285 could enhance the proliferation and metastasis of CC by up-regulating FUS, which might be a potential therapeutic target for CC treatment.

[The clinical significance of PRL-3,VEGF expression in sinonasalsquamous cell carcinoma].

Objective:To analyze the clinical significance of phosphatase of regenerating liver-3(PRL-3) and vascular endothelial growth factor(VEGF)expression in sinonasal squamous cell carcinomas.Method:We use immunohistochemical analysis and RT-PCR to detecte the expression of PRL-3 and VEGF protein in 62 cases of sinonasal squamous carcinoma tissues(SNSCC),30 cases of nasal polyps(NP),and 25 cases of normal nasal mucosa(NM).Result:①The expression of PRL-3 and VEGF in sinonasal squamous cell carcinoma tissues were statistically higher than in nasal polyps and normal nasal mucosa tissues (P<0.05).②The expression of PRL-3 and VEGF were not correlated with patient's age or gender(P>0.05).But the High expression of PRL-3 and VEGF in SNSCC was significantly related with advanced,lower tumor differentiation and lymph node metastasis(P<0.05).③The expression of PRL-3 and VEGF was positively correlated in sinonasal squamous cell carcinomas.Conclusion:The expression of PRL-3 and VEGF in SNSCC may involve in the regulation of tumor growth and differentiation.The expressing intensity may reflect the proliferation activity of sinonasal squamous cell carcinoma cell.PRL-3 and VEGF may promote the tumor metastasis in a synergistic manner.

Reaction Gas Ratio Effect on the Growth of a Diamond Film Using Microwave Plasma-Enhanced Chemical Vapor Deposition.

In this study, diamond films were prepared using the microwave plasma-enhanced chemical vapor deposition (PECVD) system, which included a DC bias system to enhance the nucleation of the films. The films were synthesized on Si wafers with different ratios of methane (CH4) and hydrogen (H2) gases. We have studied the effects of the CH4-to-H2 ratio on the structural and optical properties of diamond films. The thickness and surface profile of the films were characterized via field emission scanning electron microscopy (FE-SEM). Raman was used to investigate the structural properties of the diamond films. The refractive indexes as functions of the CH4-to-H2 ratio were measured using an ellipsometer. The FE-SEM analysis showed that the 3 and 5 sccm CH4 created diamond films. The Raman analysis indicated that a nanocrystalline diamond film was formed at 3 sccm; a general diamond film, at 5 sccm; and films similar to the a-C:H film, at 7 sccm. The ellipsometer measurement showed that the refractive index of the synthesized diamond film was around 2.42 at 3 sccm. This value decreased as the CH4 volume increased.

Dihydroartemisinin attenuates lipopolysaccharide-induced osteoclastogenesis and bone loss via the mitochondria-dependent apoptosis pathway.

Dihydroartemisinin (DHA) is a widely used antimalarial drug isolated from the plant Artemisia annua. Recent studies suggested that DHA has antitumor effects utilizing its reactive oxygen species (ROS) yielding mechanism. Here, we reported that DHA is inhibitory on lipopolysaccharide (LPS)-induced osteoclast (OC) differentiation, fusion and bone-resorption activity in vitro. Intracellular ROS detection revealed that DHA could remarkably increase ROS accumulation during LPS-induced osteoclastogenesis. Moreover, cell apoptosis was also increased by DHA treatment. We found that DHA-activated caspase-3 increased Bax/Bcl-2 ratio during LPS-induced osteoclastogenesis. Meanwhile, the translocation of apoptotic inducing factor (AIF) and the release of cytochrome c from the mitochondria into the cytosol were observed, indicating that ROS-mediated mitochondrial dysfunction is crucial in DHA-induced apoptosis during LPS-induced osteoclastogenesis. In vivo study showed that DHA treatment decreased OC number, prevents bone loss, rescues bone microarchitecture and restores bone strength in LPS-induced bone-loss mouse model. Together, our findings indicate that DHA is protective against LPS-induced bone loss through apoptosis induction of osteoclasts via ROS accumulation and the mitochondria-dependent apoptosis pathway. Therefore, DHA may be considered as a new therapeutic candidate for treating inflammatory bone loss.

[Place of clinical pharmacist in the management of drugs in patients with hypertension]. / Place du pharmacien clinicien dans la prise en charge thérapeutique du patient hypertendu.

PURPOSE: To synthesize pharmacists' interventions made in the department of internal medicine and hypertension of university hospital of Toulouse and assess the impact on medication orders. METHODS: This is a single-center, prospective study using pharmacists' interventions recorded between September 2013 and March 2014 on the Act-IP(©) website of the French Society of Clinical Pharmacy. The clinical pharmacist is present everyday in the unit to establish the medication reconciliation of new patients (the process of comparing a patient's medication orders to all of the medications that the patient has been taking), and analysis of medication orders. When a risk of iatrogenic drug is identified, a therapeutic change is proposed to the prescriber. RESULTS: A total of 2491 medication orders were analyzed for 7 months, leading to 39 pharmacists' interventions (1.6 pharmacists' interventions per 100 medication orders). The most commonly identified drug-related problems were improper administration (33%, n=13), not prescribed drug (21%, n=8), non-conformity to guidelines (18%, n=7), supratherapeutic dose (15%, n=6), and 13% (n=5) targeted prescribed treatment not administered, underdosing, incorrect administration or drug interaction. The most relevant molecules were atorvastatin (10%), bromazepam (8%) and levothyroxine (8%) and only 2 interventions targeted antihypertensive drugs. The rate of physicians' acceptance was 92%. CONCLUSION: Pharmacists' interventions mainly concern the co-prescriptions of antihypertensive drugs and very few antihypertensive drugs. The clinical pharmacist contributes to preventing iatrogenic in patients with hypertension with a very good acceptance by the clinician.

Effect of postural changes on aldosterone to plasma renin ratio in patients with suspected secondary hypertension.

AIMS: To study the influence of postural changes on aldosterone to renin ratio (ARR) in patients with suspected secondary hypertension and to evaluate the sensitivity and specificity of the recommended seated ARR compared to supine and upright ARR for primary aldosteronism screening. METHODS: Fifty-three hypertensive patients were prospectively hospitalized for secondary hypertension exploration (age: 51 ± 12, 66% males). After withdrawal of drugs interfering with renin angiotensin system, plasma aldosterone and direct renin concentration were measured in the morning, at bed after an overnight supine position, then out of bed after 1 hour of upright position and finally 2 hours later after 15 minutes of seating. Minimal renin value was set at 5 µUI/mL. RESULTS: Referring to ARR cut-off of 23 pg/µUI, the sensitivity of seated ARR was 57.1% and specificity was 92.3%. The negative and positive predictive values were 95.1% and 45.2% respectively. Compared to these results, a cut-off of 19 improved sensitivity to 85.7% with a specificity of 89.7%. Negative and positive predictive values were 98.3% and 41.1% respectively. Seated ARR mean value was lower than supine and upright ARR mean values, due to an overall increase in renin at seating compared to the supine position by factor 1.9 while aldosterone just slightly increased by factor 1.2. Seated ARR correlated to supine and upright ARR: correlation coefficients (r) 0.90 and 0.93 respectively (P<0.001). CONCLUSIONS: Current recommended measurement of ARR in the seating position is fairly correlated to supine and upright ARR. A suggested cut-off value of 19 instead of 23 pg/µUI increased the discriminating power of this test.

Mercurial sensitivity of aquaporin 1 endofacial loop B residues.

The water channel protein aquaporin-1 (AQP1) has two asparagine-proline-alanine (NPA) repeats on loops B and E. From recent structural information, these loops are on opposite sides of the membrane and meet to form a pore. We replaced the mercury-sensitive residue cysteine 189 in AQP1 by serine to obtain a mercury-insensitive template (C189S). Subsequently, we substituted three consecutive cysteines for residues 71-73 near the first NPA repeat (76-78) in intracellular loop B, and investigated whether they were accessible to extracellular mercurials. AQP1 and its mutants were expressed in Xenopus laevis oocytes, and the osmotic permeability (P(f)) of the oocytes was determined. C189S had wild-type P(f) but was not sensitive to HgCl(2). Expression of all three C189S cysteine mutants resulted in increased P(f), and all three mutants regained mercurial sensitivity. These results, especially the inhibitions by the large mercurial p-chloromercunbenzene-sulfonic acid (pCMBS) ( approximately 6A wide), suggest that residues 71-73 at the pore are accessible to extracellular mercurials. A 30-ps molecular dynamics simulation (at 300 K) starting with crystallographic coordinates of AQP1 showed that the width of the pore bottleneck (between Connolly surfaces) can vary (w(avg) = 3.9 A, sigma = 0.75; hydrated AQP1). Thus, although the pore width would be > or = 6 A only for 0.0026 of the time, this might suffice for pCMBS to reach residues 71-73. Alternative explanations such as passage of pCMBS across the AQP1 tetramer center or other unspecified transmembrane pathways cannot be excluded.

Cultured bovine corneal epithelial cells express a functional aquaporin water channel.

PURPOSE: Given recent physiological and in situ hybridization evidence for the presence of a water channel in corneal epithelium, this study was conducted to investigate its expression and characteristics using cultured bovine corneal epithelial cells (CBCEPCs). METHODS: CBCEPCs were grown in DMEM containing 2 ng/ml fibroblast growth factor and 6% fetal bovine serum. To determine their osmotic permeability (Pf), cells were passaged onto rectangular glass coverslips, and anisotonically induced volume changes were monitored by light scattering. To investigate expression, poly(A+) RNA from CBCEPCs was injected into Xenopus laevis oocytes, and the Pf of the oocytes was determined. RESULTS: For CBCEPCs challenged with a 10% hypotonic solution at 37 degrees C, the kinetic constant of volume change was k=0.52+/-0.04 seconds(-1), and the calculated Pf 72+/-6 microm/sec (n=16). The Pf of oocytes injected with water was 14+/-1.8 microm/sec (n=4); injection with poly(A+) RNA from CBCEPCs increased Pf to 77+/-6 microm/sec (n=6). This increase in Pf was inhibited by 72% (reduced to 22+/-1 microm/sec) by 0.3 mM HgCl2 and was inhibited by 56% to 58% by coinjection with aquaporin (AQP)5 antisense oligonucleotide. CONCLUSIONS: The comparatively high Pf determined for CBCEPCs, the presence of mRNA encoding water channels, and sensitivity to mercurial agents are typical of the expression of functional water channels. The predominant message is for AQP5, although the evidence was consistent with the presence of additional water channels. These findings bring renewed support for the notion that the epithelium can contribute to corneal hydration homeostasis.

Platelet activating factor inhibits fluid transport by corneal endothelium.

PURPOSE: Given reports of corneal edema after endothelial exposure to platelet activating factor (PAF), the authors have investigated whether PAF can affect the function of corneal endothelium in vitro. METHODS: The endothelial side of deepithelialized rabbit corneas was perfused with BSS+ and test agents: PAF, its inactive receptor ligand analog Lyso-PAF, and its antagonist BN52021. Stromal thickness was determined by specular microscopy. Translayer-specific electrical resistance (rho) was measured in cultured bovine corneal endothelial cells grown on permeable substrates at 36.5 degrees C. RESULTS: Control corneas perfused with BSS+ or with BSS+ containing Lyso-PAF swelled at a very slow rate (6.2 +/- 0.1, and 7.9 +/- 0.2 microns/hour, respectively). Corneas exposed to PAF swelled appreciably faster and at rates that were a saturable function of PAF (K(m), 2.1 microM); maximal rates of swelling were < 20 microns/hour, indicating no appreciable damage to intercellular junctions. BN52021 prevented PAF-induced swelling (Ki, 1.1 microM). PAF led also to a decrease in rho (from 42.8 +/- 1.4 to 24.5 +/- 0.6 omega cm2 in 1 hour; 46.8 +/- 1.5 to 38.3 +/- 1.4 omega cm2 in control layers; and 43.0 +/- 1.2 to 30.8 +/- 1.6 omega cm2 in layers exposed to PAF+BN52021). Such rho changes are consistent with swelling of intercellular spaces. CONCLUSIONS: Results suggest that PAF inhibits transendothelial fluid transport on binding to an endothelial cell receptor for it; continuous stimulation of a PAF-induced signaling cascade may lead to such inhibition. From these and other results, fluid transport might result from cascades activating sequentially basolateral and apical transporters or channels.