Experimental study on the effect and mechanism of adipose stem cell-derived exosomes combined with botulinum toxin A on skin trauma in rats.

BACKGROUND: Adipose stem cell-derived exosomes (ADSC-EXO) and botulinum toxin type A (BTX-A) individually showed a therapeutic effect on skin wound repair. AIMS: This study investigated their synergistic effect on promoting skin wound healing in vitro and in vivo and the underlying molecular events. METHODS: ADSCs were isolated from Sprague-Dawley (SD) rats to obtain ADSC-EXO by ultrafiltration and ultracentrifugation and were confirmed using nanoparticle tracking analysis and transmission electron microscopy. Human skin fibroblasts (HSF) were cultured and treated with or without ADSC-EXO, BTX-A, or their combination. Changes in cell phenotypes and protein expression were analyzed using different in vitro assays, and a rat skin wound model was used to assess their in vivo effects. RESULTS: The isolated ADSC-EXO from primarily cultured ADSCs had a circular vesicle shape with a 30-180 nm diameter. Treatment of HSF with ADSC-EXO and/or BTX-A significantly accelerated HSF migration in vitro and skin wound healing in a rat model. Moreover, ADSC-EXO plus BTX-A treatment dramatically induced VEGFA expression but reduced COL III and COL I levels in vivo. ADSC-EXO and/or BTX-A treatment significantly upregulated TGF-ß3 expression on Day 16 after surgery but downregulated TGF-ß1 expression, suggesting that ADSC-EXO plus BTX-A promoted skin wound healing and reduced inflammatory cell infiltration. CONCLUSIONS: The ADSC-EXO plus BTX-A treatment demonstrated a synergistic effect on skin wound healing through upregulation of VEGF expression and the TGF-ß3/TGF-ß1 and COL III/COL I ratio.

Identification of metabolites from edible mushroom Morchella sextelata and their biological evaluation.

To identify bioactive metabolites from the fruiting body of Morchella sextelata, fourteen metabolites (1-14) including one undescribed morchesexten A (1) were isolated. Their structures including absolute configurations were assigned on the basis of spectroscopic data and quantum chemical computational methods. Furthermore, the anti-inflammatory and antioxidant activities of the isolated compounds were evaluated. Compounds 10-12 showed inhibitory effects on nitric oxide (NO) production with IC50 values of 15.2 ± 2.7, 10.2 ± 1.9 and 35.3 ± 10.5 µM, respectively. Compounds 7 and 9 exhibited strong antioxidant effect with IC50 values of 6.7 ± 0.4 and 7.3 ± 0.8 µM compared with Vit C (IC50 15.4 ± 0.2 µM).

Construction of Exosomes that Overexpress CD47 and Evaluation of Their Immune Escape.

Our general purpose was to provide a theoretical and practical foundation for the use of exosomes (EXOs) that have high levels of CD47 as stable and efficient drug carriers. Thus, we prepared EXOs from adipose tissue-derived mesenchymal stromal cells (ADMSCs) that had high levels of CD47 (EXOsCD47) and control EXOs (without CD47), and then compared their immune escape in vivo and their resistance to phagocytosis in vitro. Nanoflow cytometry was used to determine the CD47 level in these EXOs, and the amount of EXOsCD47 that remained in rat plasma at 3 h after intraperitoneal injection. Phagocytosis of the EXOs was also determined using in vitro rat macrophage bone marrow (RMA-BM) experiments. Our in vitro results showed that macrophages ingested significantly more control EXOs than EXOsCD47 (p < 0.01), with confirmation by ultra-high-definition laser confocal microscopy. Consistently, our in vivo results showed that rats had 1.377-fold better retention of EXOsCD47 than control EXOs (p < 0.01). These results confirmed that these engineered EXOsCD47 had improved immune escape. Our results therefore verified that EXOsCD47 had increased immune evasion relative to control EXOs, and have potential for use as drug carriers.

Glycoside constituents with various antioxidant effects from fresh Cynomorium songaricum.

Cynomorium songaricum Rupr. (CSR), an edible and medicinal material, is widely cultivated in desert regions of Eastern and Western Asia, Europe, and North Africa. Ten glycoside constituents 1-10 including one new songaricumone A (1) were isolated from the fresh C. songaricum. Their structures were elucidated by comprehensive NMR data analysis. Further, various antioxidant effects of isolated compounds (1-3 and 5-10) were comprehensively and comparatively investigated. In conclusion, it is obvious that different glycosides vary significantly toward different sources of free radicals, which are attributed to different aglycones and substituted positions of sugar unit in structures.

Involvement of chemosensory proteins in host plant searching in the bird cherry-oat aphid.

Chemosensory systems are considered to play an important role in host plant selection in herbivorous insects. However, few studies have focused on chemosensory proteins (CSPs) for aphid host-location mechanisms. The roles of CSPs in searching for different Poaceae species (wheat, barley, triticale, maize and sorghum) were tested in Rhopalosiphum padi, an important cereal pest. The olfactometer assays showed that R. padi responds to plant odors. Seven R. padi CSP genes were identified. Influence of aphid morph, tissue and starvation state on expression patterns of CSPs was evaluated. Expression levels of CSP1, CSP4, CSP5 and CSP6 in winged aphids were significantly higher than those in wingless ones. Transcription levels of four genes (CSP1, CSP4, CSP5 and CSP6) were relatively higher in the head with antennae, and the four genes tended to be upregulated following starvation. Silencing of three CSPs (CSP4, CSP5 and CSP6) altered aphid host-location behavior in response to the five different host plants tested. Three volatile compounds of host plants (octanal, [E]-2-hexenol and linalool) have significant attraction to winged R. padi according to the four-arm olfactometer tests. Molecular docking predicted hydrogen bonding sites which played key roles in the binding of CSP4, CSP5 and CSP6 with volatile compounds. Knockdown of CSP4 or CSP5 significantly decreased the staying time of R. padi in the arms with octanal. However, knockdown of CSP6 could not affect the response of R. padi to octanal. These results bring evidence for the involvement of three CSPs in R. padi host-location behavior.

Functional Characterization of Two Carboxylesterase Genes Involved in Pyrethroid Detoxification in Helicoverpa armigera.

Insect carboxylesterases are major enzymes involved in metabolism of xenobiotics including insecticides. Two carboxylesterase genes, CarE001A and CarE001H, were cloned from the destructive agricultural pest Helicoverpa armigera. Quantitative real-time polymerase chain reaction showed that CarE001A and CarE001H were predominantly expressed in fat body and midgut, respectively; developmental expression analyses found that the expression levels of both CarEs were significantly higher in fifth-instar larvae than in other life stages. Recombinant CarE001A and CarE001H expressed in the Escherichia coli exhibited high enzymatic activity toward α-naphthyl acetate. Inhibition assays showed that organophosphates had strong inhibition on CarEs activity compared to pyrethroids. Metabolism assays indicated that CarE001A and CarE001H were able to metabolize ß-cypermethrin and λ-cyhalothrin. Homology modeling and molecular docking analyses demonstrated that ß-cypermethrin could fit nicely into the active pocket of both carboxylesterases. These results suggested that CarE001A and CarE001H could play important roles in the detoxification of pyrehtroids in H. armigera.

[Analysis of risk factors of ventilator-associated pneumonia in an intensive care unit].

OBJECTIVE: To explore the risk factors of ventilator-associated pneumonia (VAP) in patients admitted in an intensive care unit (ICU) for pulmonary tuberculosis (TB). METHODS: The clinical data of 143 patients admitted in the ICU at our center between January, 2014 and June, 2015 were reviewed. The patients with VAP and those without VAP were analyzed for risk factors of VAP in the setting of an ICU for pulmonary TB and compared for the duration of ventilation and hospital stay. RESULTS: The patients with pulmonary TB showed a significantly higher incidence of VAP in the ICU than those without TB. Univariate analysis suggested that the occurrence of VAP was significantly correlated with the duration of mechanical ventilation, invasive examination, pulmonary tuberculosis, lung structure changes, use of multiple antibiotics, diabetes, tracheal incision, indwelling gastric tube, APACHE II score, and coma (P<0.05). Multivariate logistic regression analysis showed that pulmonary TB, duration of mechanical ventilation, APACHE II score, invasive operation, and use of multiple antibiotics were independent risk factors for VAP (P<0.05). The patients who developed VAP had a prolonged duration of mechanical ventilation and ICU stay (P<0.05). CONCLUSION: Patients admitted in tuberculosis ICU are exposed to a high risk of VAP with a high mortality rate as the result of multiple interacting risk factors. Pulmonary TB, prolonged mechanical ventilation, an APACHE II score >15, invasive operation, and use of multiple antibiotics are all independent risk factors for VAP in tuberculosis ICU.

[The clinical significance of changes in CD4+ T cell counts by peripheral blood from patients with pulmonary tuberculosis after antitubercular treatment].

AIM: To explore the clinical significance of changes in CD4+ T cell counts by peripheral blood from patients with pulmonary tuberculosis after antitubercular treatment. METHODS: CD4+ T cell counts in the peripheral blood of 62 pulmonary TB patients were counted by flow cytometry from groups auording to the radiologic extent of disease, and compared with those obtained from 30 controls. RESULTS: (1) The baseline levels of CD4+ T cell counts (439.21+/-210.56)/mm3 in pulmonary tuberculosis group before antituberculosis therapy were significantly lower than those (748.47+/-261.85)/mm3 in the control group (P<0.01). (2) The baseline levels of CD4+ T cell counts (399.83+/-194.17)/mm3 before antituberculosis therapy in diffuse group were significantly lower than those (521.90+/-224.40)/mm3 in limited group (P<0.05). (3) After two months of antituberculosis therapy, the levels of CD4+ T cell counts in the improved group increased from (480.75+/-228.49)/mm3 to (616.75+/-280.57)/mm3, and the values were not significantly different from those in controls (P>0.05). Whereas, the levels of CD4+ T cell counts in the stable group increased from (412.97+/-197.00)/mm3 to (447.55+/-204.60)/mm3, which were still significantly lower than those in the control group (P<0.01). CONCLUSION: These results suggested that peripheral CD4+ lymphopenia was demonstrated in patients with pulmonary tuberculosis, and such lymphopenia was reversible partly with antitubercular treatment. The numbers of peripheral CD4+ T cell counts were related to the severity of the disease, the more severe of the disease was, the less numbers of the CD4+ T cell counts were.

[A retrospective study of one case of X-linked agammaglobulinemia].

OBJECTIVE: To investigate the clinical manifestations, diagnostic methods and treatment of X-linked agammaglobulinemia (XLA). METHODS: Flow cytometric analysis of the peripheral monocytes using the anti-BTK antibody was used to characterize the expression of BTK in a 21 year old male patient and his mother. The patient suffered from frequent pneumonia, and was found to be complicated with lymphocytopenia in the B cell populations, hypogammaglobulinemia (IgG 1.38 g/L, IgA 0.25 g/L, IgM 0.17 g/L) and angiotelectasis (which had not been reported in XLA patients). Sequencing of the BTK cDNA obtained from the peripheral monocytes of the patient and his mother was performed to confirm the genetic defect. RESULTS: The BTK expressions in peripheral monocytes of the patient and his mother were 96.9% and 97.8% respectively. Sequencing of the BTK gene revealed a missense mutation of R525Q in exon 16, and his mother was confirmed to be an XLA carrier. The patient was treated with immunoglobulin replacement therapy (2 g/kg). One month later, the serum IgG level of the patient was elevated to 5.79 g/L, and the clinical symptoms (included angiotelectasis), lung function and the CT scan results significantly improved. CONCLUSION: Genetic diagnosis was made for one Chinese XLA adult patient complicated with angiotelectasis. This case suggests that some XLA cases may present angiotelectasis. High dose intravenous immunoglobulin given at 2 g/kg may be of efficacy in severe XLA cases. More attention should be paid to the disease in China.

[Photoxidation reaction of Tl(I) in surface water].

The valence state of thallium affecting its toxicity, distribution and mobility, photoxidation reaction of Tl(I) was studied under the radiation from high-pressure arc mercury light or solar light. The results show that the low pH, the strong light intensity and UVB and VUC region are in favor of the photoxidation of Tl(I). In the case of pH = 2, only less than 1% Tl(I) remained in the solution after 10 min of irradiation, while pH = 9, with about 83% Tl(I) in the solution after 1 h of irradiation. After 5 min of irradiation, if the distance between the light source and the surface of solution is 20cm, just 4% Tl(I) remained in the solution, while the distance is 36 cm, still remained about 50%. 90% Tl(I) remained in filtered light, while less than 1% Tl(I) still remained in non-filtered light. The microorganic effect is not obvious comparing with photoxidation effect in this experiment, the remained Tl(I) in excluding microorganic and microorganic experiment are all about 70%.

[Comparison of paclitaxel and carboplatin versus ifosfamide, etoposide, and carboplatin regimen for treatment of patients with advanced non-small cell lung cancer].

BACKGROUND AND OBJECTIVE: The platinum-containing combination chemotherapy has been proved to be benefit to the patients with advanced non-small cell lung cancer. Compared with cisplatin-based combination chemotherapy regimens, carboplatin-based regimens result in longer survival time and less toxicity, despite of lower response rate. In the palliative treatment setting, less toxicity and longer survival may be more significant than response rate. So we choose the carboplatin-based combination chemotherapy regimens as our study objects. To compare the efficacy and toxicity of two carboplatin-based combination chemotherapy regimens: paclitaxel and carboplatin regimen(PC) vs ifosfamide, etoposide, and carboplatin regimen(IEC). METHODS: Sixty-eight patients were enrolled in this study, 35 patients received PC regimen and 33 received IEC regimen. Patients in both groups were well-matched with baseline disease characteristics(P > 0.05). RESULTS: In PC group, the response rate was 40.0% (14/35, 95% confidence interval [CI]: 23.8%-56.2%) (14 PR, 19 NC, 2 PD). Whereas in group IEC, the response rate was 21.2% (7/33, 95% CI: 7.3%-35.1%) (7 PR, 24 NC, 2 PD). The median survival and 1-year survival rate were 9.1 months (95% CI: 7.2-11.0 months) and 25.7% (95% CI: 11.2%-40.2%) for group PC versus 7.8 months (95% CI: 6.2-9.4 months) and 20.0% (95% CI: 6.0%-34.0%) for group IEC. The better response rate, 1-year survival rate, and the longer median survival seen in the group PC were not statistically significant (for response rate, P = 0.094, Chi-square test; for overall survival, P = 0.684, Log-rank test). The hematologic toxicities, especially leukopenia (P < 0.0005, Wilcoxon rank sum test) and anemia (P = 0.006, Wilcoxon rank sum test) were less pronounced in group PC; Hematuria and fever were pronounced more in group IEC, whereas allergic reaction was more in group PC, but there were no statistically significant differences between the two groups(P > 0.05, Wilcoxon rank sum test); Other toxicities were similar. CONCLUSIONS: Compared with IEC regimen, PC regimen result in less hematologic toxicities in the patients with advanced NSCLC, Although whether its efficacy was better should be confirmed by well-controlled randomized clinical trials with more patients.