Surviving winter on the Qinghai-Tibetan Plateau: Pikas suppress energy demands and exploit yak feces to survive winter.

The Qinghai-Tibetan Plateau, with low precipitation, low oxygen partial pressure, and temperatures routinely dropping below -30 °C in winter, presents several physiological challenges to its fauna. Yet it is home to many endemic mammalian species, including the plateau pika (Ochotona curzoniae). How these small animals that are incapable of hibernation survive the winter is an enigma. Measurements of daily energy expenditure (DEE) using the doubly labeled water method show that pikas suppress their DEE during winter. At the same body weight, pikas in winter expend 29.7% less than in summer, despite ambient temperatures being approximately 25 °C lower. Combined with resting metabolic rates (RMRs), this gives them an exceptionally low metabolic scope in winter (DEE/RMRt = 1.60 ± 0.30; RMRt is resting metabolic rate at thermoneutrality). Using implanted body temperature loggers and filming in the wild, we show that this is achieved by reducing body temperature and physical activity. Thyroid hormone (T3 and T4) measurements indicate this metabolic suppression is probably mediated via the thyroid axis. Winter activity was lower at sites where domestic yak (Bos grunniens) densities were higher. Pikas supplement their food intake at these sites by eating yak feces, demonstrated by direct observation, identification of yak DNA in pika stomach contents, and greater convergence in the yak/pika microbiotas in winter. This interspecific coprophagy allows pikas to thrive where yak are abundant and partially explains why pika densities are higher where domestic yak, their supposed direct competitors for food, are more abundant.

Bacillus velezensis strain NA16 shows high poultry feather-degrading efficiency, protease and amino acid production.

Isolated Bacillus velezensis strain NA16, which produces proteases, amino acids and the transcription levels of different keratinolytic enzymes and disulfide reductase genes in whole gene sequencing, was evaluated during feather degradation. The result shows under optimum fermentation conditions, chicken feather fermentation showed total amino acid concentration of 7599 mg/L, degradation efficiency of 99.3% at 72 h, and protease activity of 1058 U/mL and keratinase activity of 288 U/mL at 48 h. Goose feather fermentation showed total amino acid concentration of 4918 mg/L (96 h), and degradation efficiency was 98.9% at 120 h. Chicken feather fermentation broth at 72 h showed high levels of 17 amino acids, particularly phenylalanine (1050 ± 1.90 mg/L), valine (960 ± 1.04 mg/L), and glutamic (950 ± 3.00 mg/L). Scanning electron microscopy and Fourier transform infrared analysis revealed the essential role of peptide bond cleavage in structural changes and degradation of feathers. Protein purification and zymographic analyses revealed a key role in feather degradation of the 39-kDa protein encoded by gene1031, identified as an S8 family serine peptidase. Whole genome sequencing of NA16 revealed 26 metalloproteinase genes and 22 serine protease genes. Among the proteins, S8 family serine peptidase (gene1031, gene1428) and S9 family peptidase (gene3132) were shown by transcription analysis to play major roles in chicken feather degradation. These findings revealed the transcription levels of different families of keratinolytic enzymes in the degradation of feather keratin by microorganisms, and suggested potential applications of NA16 in feather waste management and amino acid production.

Molecular Dynamics Study on Regimes of Head-on Droplet Collision.

The head-on collision of two water droplets with a diameter of 10 nm in an atmospheric environment was investigated via molecular dynamics simulations. The gas molecules between droplets were visualized, and the phenomena of gas extrusion and gas molecules being captured were found. By observing and analyzing the holes regime, a "periphery-sucking" mechanism was proposed to explain the thinning in the middle of the expanding disk and the holes appearing. It was found that the splattering regime can be divided into the limited splattering regime and the divergent splattering regime. The splattering modes and droplet characteristics of the two regimes are markedly different. The non-bonded interactions and intermolecular hydrogen bond were analyzed, and it was found that increasing the Weber number (We) can effectively promote the mixing of the two droplets, promote the formation of an intermolecular hydrogen bond between the two droplets, and reduce the average lifetime of the intermolecular hydrogen bond. The radial distribution function between the water molecules was plotted, showing that increasing the We makes the water molecules more dispersed as a whole in the collision process.

Injection therapies for lateral epicondylalgia: a systematic review and Bayesian network meta-analysis.

BACKGROUND: There are many injection therapies for lateral epicondylalgia but there has been no previous comprehensive comparison, based on the Bayesian method. METHODS: The MEDLINE, EMBASE and the Cochrane Central Register of Controlled Trials (CENTRAL) databases were searched for appropriate literature. The outcome measurement was the pain score. Direct comparisons were performed using the pairwise meta-analysis, and network meta-analysis, based on a Bayesian model, was used to calculate the results of all of the potentially possible comparisons and rank probabilities. A sensitivity analysis was performed by excluding low-quality studies. The inconsistency of the model was assessed by means of the node-splitting method. Metaregression was used to assess the relationship between the sample size and the treatment effect. RESULTS: All of the injection treatments showed a trend towards better effects than placebo. Additionally, the peppering technique did not add additional benefits when combined with other treatments. No significant changes were observed by excluding low-quality studies in the sensitivity analysis. No significant inconsistencies were found according to the inconsistency analysis, and metaregression revealed that the sample size was not associated with the treatment effects. CONCLUSIONS: Some commonly used injection therapies can be considered treatment candidates for lateral epicondylalgia, such as botulinum toxin, platelet-rich plasma and autologous blood injection, but corticosteroid is not recommended. Hyaluronate injection and prolotherapy might be more effective, but their superiority must be confirmed by more research. The peppering technique is not helpful in injection therapies.

Molecular Mechanism of Calycosin Inhibited Vascular Calcification.

Vascular calcification (VC) is a pathological condition frequently observed in cardiovascular diseases. Primary factors contributing to VC are osteogenic differentiation of vascular smooth muscle and hydroxyapatite deposition. Targeted autophagy (a lysosome-mediated mechanism for degradation/recycling of unnecessary cellular components) is a useful approach for inhibiting VC and promoting vascular cell health. Calycosin has been shown to alleviate atherosclerosis by enhancing macrophage autophagy, but its therapeutic effect on VC has not been demonstrated. Using an in vitro model (rat thoracic aortic smooth muscle cell line A7r5), we demonstrated effective inhibition of VC using calycosin (the primary flavonoid component of astragalus), based on the enhancement of autophagic flux. Calycosin treatment activated AMPK/mTOR signaling to induce initiation of autophagy and restored mTORC1-dependent autophagosome-lysosome fusion in late-stage autophagy by promoting soluble N-ethylmaleimide-sensitive factor attachment protein receptor (SNARE) complex formation, thereby preventing stoppage of autophagy in calcified cells. Calycosin substantially reduced degrees of both osteogenic differentiation and calcium deposition in our VC cell model by enhancing autophagy. The present findings clarify the mechanism whereby calycosin mitigates autophagy stoppage in calcified smooth muscle cells and provide a basis for effective VC treatment via autophagy enhancement.

RhB@MOF-5 Composite Film as a Fluorescence Sensor for Detection of Chilled Pork Freshness.

This study presents a novel composite thin film based on rhodamine B encapsulated into MOF-5 (Metal Organic Frameworks) as a fluorescence sensor for the real-time detection of the freshness of chilled pork. The composite film can adsorb and respond to the volatile amines produced by the quality deterioration of pork during storage at 4 °C, with the fluorescence intensity of RhB decreasing over time. The quantitative model used for predicting the freshness indicator (total volatile base nitrogen) of pork was built using the fluorescence spectra (excited at 340 nm) of the RhB@MOF-5 composite film combined with the partial least squares (PLS) algorithm, providing Rc2 and Rp2 values of 0.908 and 0.821 and RMSEC (root mean square error of calibration) and RMSEP (root mean square error of prediction) values of 3.435 mg/100 g and 3.647 mg/100 g, respectively. The qualitative model established by the partial least squares discriminant analysis (PLS-DA) algorithm was able to accurately classify pork samples as fresh, acceptable or spoiled, and the accuracy was 86.67%.

Comparison of the immersion chilling and freezing and traditional air freezing on the quality of beef during storage.

Compared to traditional air freezing, immersion chilling and freezing shows an improvement in the freezing effect on meat quality, but it is not known whether this advantage persists over longer storage periods. Therefore, the objective of the current study was to compare the effects of immersion chilling and freezing (ICF) and traditional air freezing (TAF) on the physical and chemical indexes in beef longissimus muscle during a storage period of 150 days. In the current study, the longissimus muscle from Luxi cattle (aged 20-24 months) was analyzed, with samples independently frozen by ICF and TAF. After the core temperature was frozen to below -18 degrees by the two chilling methods, samples were transferred to a -18 degrees cold room for further storage. During the storage period, physical and chemical indexes, mainly including color and texture qualities, total volatile base nitrogen (TVB-N) and peroxide value (POV) were measured and comparatively analyzed at several fixed time points. A higher freezing rate was observed in ICF (5.124 cm/h) than in TAF (0.194 cm/h), and better microstructure was observed in ICF treatment. Besides, peak force values and total energy values were significantly lower in the TAF group than in the ICF group during the first 45 days of freezing storage time (p < .05). ICF also showed better color quality due to higher L* values than TAF samples during the first 75 days of frozen storage (p < .05). In addition, the thawing loss (after 75 days of storage), total volatile base nitrogen, and peroxide value (in the 30 to 75 days of storage period) were lower in the ICF than in the TAF group. In conclusion, the immersion chilling and freezing is more conducive to the quality of beef during storage at -18 degrees compared to traditional air freezing.

Novel ß-mannanase/GLP-1 fusion peptide high effectively ameliorates obesity in a mouse model by modifying balance of gut microbiota.

We constructed a novel ß-mannanase/GLP-1 fusion peptide, termed MGLP_1, and evaluated its ability to ameliorate obesity in a high-fat/high-sugar diet (HFSD)-induced mouse model. Eight-wk MGLP_1 treatment notably reduced obesity, as reflected by significant changes of body weight, serum triglyceride level, fatty liver and adipose tissue distribution. Amelioration of HFSD-induced gut dysbiosis by MGLP_1 was evidenced by reduced abundance ratio of bacterial phyla Firmicutes to Bacteroidetes, enhanced abundance of beneficial probiotic genera (Bifidobacterium, Lachnospiraceae, Ileibacterium), and reduced abundance of harmful genera (Clostridium, Romboutsia). Mechanisms of weight loss were investigated by comparing effects of treatment with MGLP_1 vs. prebiotics manno-oligosaccharides (MOS). MGLP_1 ameliorated gut microbiota imbalance by enhancing carbohydrate catabolism, whereas MOS promoted glycan synthesis and metabolism. Our findings, taken together, indicate that MGLP_1 fusion peptide has strong potential for amelioration of obesity by modifying relationships between gut microbiota and lipid and glucose metabolism.

A Microfluidic Biosensor Based on Magnetic Nanoparticle Separation, Quantum Dots Labeling and MnO2 Nanoflower Amplification for Rapid and Sensitive Detection of Salmonella Typhimurium.

Screening of foodborne pathogens is an effective way to prevent microbial food poisoning. A microfluidic biosensor was developed for rapid and sensitive detection of Salmonella Typhimurium using quantum dots (QDs) as fluorescent probes for sensor readout and manganese dioxide nanoflowers (MnO2 NFs) and as QDs nanocarriers for signal amplification. Prior to testing, amino-modified MnO2 nanoflowers (MnO2-NH2 NFs) were conjugated with carboxyl-modified QDs through EDC/NHSS method to form MnO2-QD NFs, and MnO2-QD NFs were functionalized with polyclonal antibodies (pAbs) to form MnO2-QD-pAb NFs. First, the mixture of target Salmonella Typhimurium cells and magnetic nanoparticles (MNPs) modified with monoclonal antibodies (mAbs) was injected with MnO2-QD-pAb NFs into a microfluidic chip to form MNP-bacteria-QD-MnO2 complexes. Then, glutathione (GSH) was injected to dissolve MnO2 on the complexes into Mn2+, resulting in the release of QDs. Finally, fluorescent intensity of the released QDs was measured using the fluorescent detector to determine the amount of Salmonella. A linear relationship between fluorescent intensity and bacterial concentration from 1.0 × 102 to 1.0 × 107 CFU/mL was found with a low detection limit of 43 CFU/mL and mean recovery of 99.7% for Salmonella in spiked chicken meats, indicating the feasibility of this biosensor for practical applications.

[Effects of epidermal growth factor receptor on airway inflammation in a rat asthmatic model].

OBJECTIVE: To explore the possible roles of epidermal growth factor receptor (EGFR) in the process of acute and chronic airway inflammation in a rat asthmatic model. METHODS: Forty-five Sprague-Dawley (SD) rats were randomly divided into control groups (subgroups A1, A2, A4), asthmatic groups (subgroups B1, B2, B4) and treatment groups (subgroups C1, C2, C4), with 5 mice in each subgroup. Mice in the asthmatic and treatment groups were exposed to OVA challenge for 1 week, 2 weeks and 4 weeks. Rats in the treatment groups received intraperitoneal injection of a tyrosine kinase inhibitor Genistein (Rongli China) with the dose of 20 mg/kg 1 hour before OVA exposure. Total cell counts and cell differentials in bronchoalveolar lavage fluid (BALF) were performed. A semi-quantified method of airway inflammation score was used to evaluate airway inflammation by hematoxylin-eosin (HE) staining. Expression of EGFR and tyrosine phosphorylation (EGFR activation) in airway epithelium at different times of OVA exposure were evaluated by immunohistochemical and immunofluorescence. All data were expressed as mean +/- SD. One-way ANOVA was used for comparison between 2 groups and post-hoc multiple comparisons of means were performed by using Least Significant Difference. RESULTS: (1) The total cell counts and cell differentials in the BALF of subgroups B1, B2 and B4 were higher than those of subgroups A1, A2 and A4. The total cell counts and eosinophils (EOS) in the BALF of subgroups C1, C2, and C4 [Total cells (48 +/- 6) x 10(5), (51 +/- 9) x 10(5), (57 +/- 12) x 10(5); EOS (2.5 +/- 0.5) x 10(5), (2.7 +/- 0.6) x 10(5), (2.6 +/- 0.5) x 10(5), respectively] decreased significantly as compared to those of subgroups B1, B2 and B4 [Total cells (70 +/- 10) x 10(5), (88 +/- 8) x 10(5), (72 +/- 10) x 10(5); EOS (5.6 +/- 0.8) x 10(5), (6.6 +/- 0.6) x 10(5), (4.3 +/- 0.4) x 10(5)], all P < 0.05. There was no significant difference in the counts of neutrophils and lymphocytes in BALF between the treatment groups and the asthmatic groups. The count of epithelial cells in group C1 [(2.5 +/- 0.5) x 10(5)] was lower than that in group B1[(4.9 +/- 0.7) x 10(5)], q = 4.671, P < 0.05. But that in group C4[(5.7 +/- 1.2) x 10(5)] was higher than that in group B4 [(4.3 +/- 0.4) x 10(5)], q = 4.012, P < 0.05. (2) The airway inflammation score in group C4(3.6 +/- 0.6) was less than that in group B4(5.1 +/- 0.6), q = 4.923, P < 0.05. The scores of group C1 and C2 were less than those of group B1 and B2, but the differences did not reach statistical significance. (3) The expression of EGFR and tyrosine phosphorylation in airway epithelium of the OVA sensitized subgroups were increased statistically as compared to the control subgroups (all P < 0.05). Genistein decreased tyrosine phosphorylation of EGFR in subgroups C1, C2 and C4[(3.12 +/- 0.24), (3.00 +/- 0.28), (2.69 +/- 0.54)] as compared to subgroups B1, B2 and B4[(3.69 +/- 0.43), (3.57 +/- 0.29), (4.46 +/- 0.47), respectively] (all P < 0.05). (4) There were positive correlations between expression and activation of EGFR in airway epithelium and total cell counts, EOS counts, neutrophil and lymphocyte counts in BALF, and airway inflammation scores (all P < 0.05). CONCLUSIONS: EGFR is involved in airway inflammation of asthmatic rats. Tyrosine kinase inhibitor Genistein inhibits acute and chronic airway inflammation in the asthmatic model.

Effect of interleukin-5 receptor-alpha short hairpin RNA-expressing vector on bone marrow eosinophilopoiesis in asthmatic mice.

Bone marrow eosinophilopoiesis induced by interleukin (IL)-5 is a major contributor to eosinophilic airway inflammation in asthma. However,research on the use of IL-5 receptor alpha (IL-5Ralpha) as the target has seldom been reported. This study was undertaken to explore the effects of inhibition of IL-5Ralpha expression through an IL-5Ralpha short hairpin RNA-expressing vector on bone marrow eosinophilopoiesis and airway inflammation in an asthmatic mouse model. An effective plasmid vector was selected that could express short hairpin RNA targeted at IL-5Ralpha (P-IL-5Ralpha). An adenovirus vector (Ad) was then constructed that was inserted in an effective template sequence (Ad-IL-5Ralpha). An animal model of asthma was established by sensitizing and challenging Balb/c mice with ovalbumin. Animals were treated intravenously with Ad-IL-5Ra and changes in bone marrow eosinophilopoiesis and airway inflammation were detected in asthmatic mice. Investigators found that P-IL-5Ra-3 targeted at the sequence of CAG CTG CCT GGT TCG TCT T markedly suppressed IL-5Ralpha expression in B lymphoma cells in vitro. In addition, Ad-IL-5Ralpha could suppress IL-5Ralpha expression in murine bone marrow cells in vitro and in vivo, and it could significantly decrease IL-5-induced eosinophilia in cultured bone marrow cells. Additional studies indicated that intravenously injected Ad-IL-5Ralpha not only selectively reduced the number of eosinophils in the bone marrow, peripheral blood, and bronchoalveolar lavage fluid, it also relieved airway inflammation in asthmatic mice. Results reported here show that blocking of IL-5Ralpha expression through RNA interference can enhance effective treatment of asthma, and that bone marrow can be used as a key targeted organ in the treatment of asthmatic mice.

Effects of Astragalus membranaceus in promoting T-helper cell type 1 polarization and interferon-gamma production by up-regulating T-bet expression in patients with asthma.

OBJECTIVE: To explore the effect of Astragalus membranaceus (AM) on T-helper cell type 1 (Thl) specific transcription factor T-box expressed in T cells (T-bet) expression and Thl/Th2 equilibrium. METHODS: The levels of T-bet mRNA in peripheral blood mononuclear cells (PBMCs) from 15 patients with asthma and 15 healthy subjects were determined by reverse transcription-polymerase chain reaction (RT-PCR). PBMCs in asthma patients were incubated with AM and then the concentration of interferon gamma (IFN-gamma) and interleukin-4 (IL-4) in the supernate before and after AM intervention were determined by ELISA. The numbers of CD4 + CCR3 + and CD4 + CCR5 + cells were counted by flow cytometry. RESULTS: The expression of T-bet mRNA and the level of IFN-gamma were lower, but level of serum IL-4 was higher in asthma patients when compared with those in healthy subjects respectively. After AM (60 microg/ml) intervention, the former two parameters raised and showed a positive correlation between them, while the level of IL-4 was decreased. The mean percentage of CD4 + CCR3 + cells in asthma patients was significantly higher but that of CD4 + CCR5 + cells was lower when compared with those in healthy subjects respectively. After AM intervention, the abnormal change in the two indexes was improved to certain extent, showing a reversing status of Th2 polarization. CONCLUSION: AM could increase the expression of T-bet mRNA and Thl cytokines such as IFN-Y, and might reverse the Th2 predominant status in asthma patients.

Circulating ghrelin in patients with chronic obstructive pulmonary disease.

OBJECTIVE: Unexplained weight loss is common in patients with chronic obstructive pulmonary disease (COPD). Because ghrelin plays an important role in energy homeostasis, this study investigated the plasma level of ghrelin in COPD. METHODS: Plasma ghrelin levels and levels of leptin, tumor necrosis factor-alpha, and C-reactive protein were measured in 29 patients with COPD and 17 healthy controls. Body composition was assessed with bioelectrical impedance analysis. RESULTS: Body mass index and percentage of body fat were lower in patients who had COPD than in healthy controls. Plasma ghrelin and leptin concentrations were significantly lower in patients who had COPD than in healthy controls (ghrelin: 0.25+/-0.22 ng/mL versus 0.43+/-0.24 ng/mL, P=0.013; leptin: 1.77+/-0.70 ng/mL versus 2.85+/-0.96 ng/mL, P=0.000). In contrast, tumor necrosis factor-alpha and C-reactive protein were significantly higher in those with COPD than in controls. Plasma ghrelin (log transformed) was positively correlated with body mass index and percentage of body fat in patients with COPD but negatively correlated in control subjects. Plasma ghrelin was negatively correlated with tumor necrosis factor-alpha and C-reactive protein in COPD. CONCLUSION: Plasma ghrelin level was decreased in COPD and this is different from other weight-loss diseases. These data suggest that decreased ghrelin and other factors may contribute to alterations in metabolic status during inflammatory stress in this disease.

STAT1 antisense oligonucleotides attenuate the proinflammatory cytokine release of alveolar macrophages in bleomycin-induced fibrosis.

To investigate the effect of signal transducers and activators of transcription 1 (STAT1) antisense oligonucleotides (ASON) on concentrations of TNF-alpha, IL-8, NO secreted by alveolar macrophages (AMs) in bleomycin-induced rat pulmonary fibrosis, five adult female Wistar rats were intratracheally instilled with bleomycin. After 7 days, the rats were killed by right ventricle of heart exsanguinations under ketamine anaesthesia and bronchoalveolar lavage (BAL) was performed to obtain AMs. AMs were divided into four groups, treated with STAT1 ASON, STAT1 sense oligonucleotides (SON), dexamethasone (DEX) and medium alone (control), respectively. AMs and media were collected after culture for 36 h. The mRNA and protein expressions of STAT1 and ICAM-1 in AMs were detected by RT-PCR and ELISA, respectively. The concentrations of TNF-alpha, IL-8, NO in cultured medium were detected. The STAT1 mRNA expression by AMs in the STAT1 ASON group was lower than those of AMs in the STAT1 SON group, the DEX group and the control group (p<0.05). Moreover, the STAT1 mRNA expression by AMs in the DEX group was also lower than those of AMs in the STAT1 SON group and the control group (p<0.05), but the STAT1 mRNA expression by AMs in the STAT1 SON group was not different from that of the control group (p>0.05). The protein expressions of STAT1 and ICAM-1 and the mRNA expression of ICAM-1 showed similar changes to the STAT1 mRNA expression by AMs. The concentrations of TNF-alpha, IL-8, NO in cultured medium from STAT1 ASON group were lower than those from STAT1 SON, DEX and the control groups (p<0.05). Moreover, the concentrations of TNF-alpha, IL-8, NO in cultured medium from DEX group were also lower than those from the control and STAT1 SON group (p<0.05), but no difference between STAT1 SON group and the control (p>0.05). The results suggest that STAT1 ASON could inhibit the secretion of TNF-alpha, IL-8, NO in AMs, and STAT1 could become a target of treating pulmonary fibrosis.

Relationship between bone mineral density and syndrome types described in traditional chinese medicine in chronic obstructive pulmonary disease: a preliminary clinical observation.

Osteoporosis is a common finding following chronic obstructive pulmonary disease (COPD), but there are few reports on the relationship between bone mineral density (BMD) and the syndrome types described in traditional Chinese medicine (TCM) in patients with COPD. A cross-sectional medical survey was used in this study. Twenty-six male patients with COPD and 26 age-matched male healthy subjects were recruited. The symptom questionnaire survey of TCM was implemented, and thereafter the COPD patients were divided into two subgroups: type of deficiency of the lung and spleen (TDLS) and type of deficiency of the lung, spleen and kidney (TDLSK). BMD of lumbar spine (L2-4), non-dominant femoral neck (Neck), Ward's triangle (Ward's), and great trochanter (Troch) were measured by dual-energy x-ray absorptiometry. In addition, the other bone turnover markers were also examined. The results showed that BMD was much more decreased in TDLSK than that in TDLS patients (p < 0.05), and BMD in the patients of the TDLS subgroup without symptoms of kidney-vacuity has showed the decreased trend from healthy subjects to TDLS patients. Furthermore, there was a higher incidence of osteoporosis in patients with TDLSK compared with that in TDLS (p < 0.05, OR > 2.0). Therefore, the data suggest that: (1) BMD might be a marker more sensitive than the symptom for the diagnosis of kidney-vacuity in COPD patients; (2) the deficiency of kidney would be the key factor of bone mineral loss; and (3) that invigorating the kidney should be performed in the phase of TDLS in COPD patients in advance.

[Simvastatin induces eosinophil apoptosis in vitro].

OBJECTIVE: To investigate the effect of simvastatin, a 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR) inhibitor, on eosinophils (EOSs) apoptosis in asthma patients. METHODS: Peripheral blood EOSs from 10 asthma patients were cultured in the presence or absence of simvastatin (1, 5, 10, 20 micromol/L), together with or without mevalonate (100 micromol/L) for 6, 12, 24, and 48 h. Apoptosis was monitored by annexin V/PI staining and flow cytometry. Caspase-3 was measured by enzyme-linked immunosorbent assay (ELISA). RESULTS: EOSs were particularly susceptible to apoptosis after incubated with 5 micromol/L simvastatin for 6, 12, 24, and 48 h [the rates of EOSs undergoing apoptosis were: (23 +/- 3)%, (24 +/- 3)%, (41 +/- 6)%, (70 +/- 12)% in control and (32 +/- 4)%, (47 +/- 7)%, (62 +/- 9)%, (86 +/- 14)% in simvastatin; compared with control at the same time point: P = 0.000]. EOS apoptosis occurred at doses of 1 micromol/L and was already maximal at 5 micromol/L after incubated with simvastatin for 12 h [the rates of EOSs undergoing apoptosis were: (24 +/- 3)% in control, (37 +/- 3)%, (51 +/- 3)%, (53 +/- 4)%, (52 +/- 4)% in 1, 5, 10, 20 micromol/L simvastatin, respectively; compared with control: P = 0.000]. The level of caspase-3 in EOSs was consistent with the rate of cell apoptosis [(8 +/- 3) microg/L in control, (14 +/- 4), (22 +/- 4), (24 +/- 4), (23 +/- 5) microg/L in 1, 5, 10, 20 micromol/L simvastatin, respectively; compared with control: P = 0.000 - 0.003]. However, Co-incubation of simvastatin with mevalonate (the production of HMGR) completely reversed the activity of simvastatin on EOS apoptosis even when the highest simvastatin (20 micromol/L) dose was used; the rates of EOSs undergoing apoptosis in the control, mevalonate plus simvastatin and simvastatin alone were (24 +/- 3)%, (52 +/- 4)% and (25 +/- 3)%, respectively; while the caspase-3 levels were (8 +/- 3) microg/L, (23 +/- 5) microg/L and (9 +/- 3) microg/L, respectively. CONCLUSION: Simvastatin induces apoptosis of EOSs in asthma patients via its ability to block the synthesis of the important isoprenoid intermediates, which leads to the inhibition of small GTP-binding protein activity.

[The effect of STAT1 antisense oligonucleotides on lung fibroblast proliferation and hydroxyproline secretion].

OBJECTIVE: To investigate the effect of signal transducer and activator of transcription 1 (STAT1) antisense oligonucleotides on lung fibroblast proliferation and hydroxyproline secretion. METHODS: Ten adult female Wistar rats were randomly divided into two groups: one group was intratracheally instilled with bleomycin (BLM), while another group with 0.9% NaCl solution (NS). After 7 days, the rats were killed by right ventricle of heart exsanguinations under ketamine anaesthesia, and bronchoalveolar lavage (BAL) was performed to obtain alveolar macrophage (AM). AMs from the BLM group were divided into four groups, treated with STAT1 antisense oligonucleotides, STAT1 sense oligonucleotides, dexamethasone and medium alone (control), respectively. AMs and media were collected after culture for 36 h. The mRNA and protein expression of STAT1 and ICAM-1 in AMs were detected by RT-PCR and Cell-ELISA, respectively. The conditioned media were co-cultured with lung fibroblasts for 30 h, and then the cell proliferation and the concentration of hydroxyproline were examined. RESULTS: (1) The STAT1 mRNA expression by AMs in the STAT1 antisense oligonucleotides group (31.8 +/- 3.5) was lower than those of AMs in the STAT1 sense oligonucleotides group (64.2 +/- 4.3), the dexamethasone group (44.1 +/- 4.6) and the control group (65.5 +/- 4.6) (P < 0.05). Moreover, the STAT1 mRNA expression by AMs in the dexamethasone group was also lower than those of AMs in the STAT1 sense oligonucleotides group and the control group (P < 0.05), but the STAT1 mRNA expression by AMs in the STAT1 sense oligonucleotides group was not different from that of the control group (P > 0.05). The STAT1 mRNA expression by AMs in the NS group (14.9 +/- 3.1) was lower than those of AMs in the STAT1 antisense oligonucleotides group, the STAT1 sense oligonucleotides group, the dexamethasone group and the control group (P < 0.05). (2) The mRNA expression of ICAM-1 showed similar changes to the STAT1 mRNA expression by AMs. (3) The STAT1 protein expression by AMs in the STAT1 antisense oligonucleotides group (4.4 +/- 0.6) or in the NS group (3.7 +/- 0.4) was lower than those of AMs in the STAT1 sense oligonucleotides group (7.7 +/- 0.7), the dexamethasone group (5.9 +/- 0.4) and the control group (7.6 +/- 0.6) (P < 0.05); and the STAT1 protein expression by AMs in the dexamethasone group was also lower than those of AMs in the STAT1 sense oligonucleotides group and the control group (P < 0.05), but the STAT1 protein expression by AMs in the STAT1 sense oligonucleotides group was not different from that of the control group (P > 0.05). (4) The changes of ICAM-1 protein expression, lung fibroblast proliferation and hydroxyproline concentration were consistent with the changes of STAT1 protein expression by AMs. CONCLUSIONS: STAT1 antisense oligonucleotides could inhibit the mRNA and the protein expression of STAT1 and ICAM-1 in AMs. STAT1 antisense oligonucleotides also inhibited lung fibroblast proliferation and hydroxyproline secretion.

[Effect of tyrosine kinase inhibitors on airway remodeling in bronchial asthma].

OBJECTIVE: To investigate the effect of tyrosine kinase inhibitors (TKIs) on asthmatic rat airway remodeling. METHODS: The inhibitive effects of three TKIs (Genistein, jin-zhuan-ting and Tyrphostin AG1478) on proliferation of primary cultures of rat tracheal epithelial cells were assessed by MTT assay. Then, jin-zhuan-ting was adopted in the asthmatic rat model; immunohistofluorescene method was used to stain phosphorylated tyrosine (P-tyr) for disclosing the activation of EGFR; Sirius Red staining of submucosal collagen I and III was performed, and an analysis was made on the correlation between EGFR activation and collagen I and III deposition. RESULTS: All the three TKIs inhibited the growth of tracheal epithelial cells in a time and dose depending manner, and the inhibition rates among them showed no statistical differences; airway subepithelial collagen I and III deposition degrees were markedly elevated in asthmatic groups and jin-zhuan-ting reduced the deposition in a certain degree; EGFR activation (P-tyr) in airways epithelium of asthmatic groups was greatly increased in comparison with that of control groups, and it was evidently decreased in jin-zhuan-ting groups. Correlation analysis demonstrated that the amount of airway subepithelial collagen I and III was positively correlated to EGFR activation. CONCLUSION: TKIs may have preventive implications for asthmatic airway remodeling.

Treatments for shoulder impingement syndrome: a PRISMA systematic review and network meta-analysis.

Many treatments for shoulder impingement syndrome (SIS) are available in clinical practice; some of which have already been compared with other treatments by various investigators. However, a comprehensive treatment comparison is lacking. Several widely used electronic databases were searched for eligible studies. The outcome measurements were the pain score and the Constant-Murley score (CMS). Direct comparisons were performed using the conventional pair-wise meta-analysis method, while a network meta-analysis based on the Bayesian model was used to calculate the results of all potentially possible comparisons and rank probabilities. Included in the meta-analysis procedure were 33 randomized controlled trials involving 2300 patients. Good agreement was demonstrated between the results of the pair-wise meta-analyses and the network meta-analyses. Regarding nonoperative treatments, with respect to the pain score, combined treatments composed of exercise and other therapies tended to yield better effects than single-intervention therapies. Localized drug injections that were combined with exercise showed better treatment effects than any other treatments, whereas worse effects were observed when such injections were used alone. Regarding the CMS, most combined treatments based on exercise also demonstrated better effects than exercise alone. Regarding surgical treatments, according to the pain score and the CMS, arthroscopic subacromial decompression (ASD) together with treatments derived from it, such as ASD combined with radiofrequency and arthroscopic bursectomy, showed better effects than open subacromial decompression (OSD) and OSD combined with the injection of platelet-leukocyte gel. Exercise therapy also demonstrated good performance. Results for inconsistency, sensitivity analysis, and meta-regression all supported the robustness and reliability of these network meta-analyses. Exercise and other exercise-based therapies, such as kinesio taping, specific exercises, and acupuncture, are ideal treatments for patients at an early stage of SIS. However, low-level laser therapy and the localized injection of nonsteroidal anti-inflammatory drugs are not recommended. For patients who have a long-term disease course, operative treatments may be considered, with standard ASD surgery preferred over arthroscopic bursectomy and the open surgical technique for subacromial decompression. Notwithstanding, the choice of surgery should be made cautiously because similar outcomes may also be achieved by the implementation of exercise therapy.