Modulation of gut microbiota alleviates cerebral ischemia/reperfusion injury in rats by inhibiting M1 polarization of microglia.

Gut microbiota affects the gut-brain axis; hence, the modulation of the microbiota has been proposed as a potential therapeutic strategy for cerebral ischemia/reperfusion injury (CIRI). However, the role and mechanism of the gut microbiota in regulating microglial polarization during CIRI remain poorly understood. Herein, using a middle cerebral artery occlusion and reperfusion (MCAO/R) rat model, we evaluated changes in the gut microbiota after CIRI and the potential effects of fecal microbiota transplant (FMT) on the brain. Rats underwent either MCAO/R or sham surgery, and then they received FMT (started 3 days later; continued for 10 days). 2,3,5-Triphenyltetrazolium chloride staining, neurological outcome scale, and Fluoro-Jade C staining showed that MCAO/R induced cerebral infarction, neurological deficits, and neuronal degeneration. In addition, immunohistochemistry or real-time PCR assay showed increased expression levels of M1-macrophage markers-TNF-α, IL-1ß, IL-6, and iNOS-in the rats following MCAO/R. Our finding suggests that microglial M1 polarization is involved in CIRI. 16 S ribosomal RNA gene sequencing data revealed an imbalance in the gut microbiota of MCAO/R animals. In contrast, FMT reversed this MCAO/R-induced imbalance in the gut microbiota and ameliorated nerve injury. In addition, FMT prevented the upregulation in the ERK and NF-κB pathways, which reversed the M2-to-M1 microglial shift 10 days after MCAO/R injury in rats. Our primary data showed that the modulation of the gut microbiota can attenuate CIRI in rats by inhibiting microglial M1 polarization through the ERK and NF-κB pathways. However, an understanding of the underlying mechanism requires further study.

Comparative transcriptome and metabolome analyses identified the mode of sucrose degradation as a metabolic marker for early vegetative propagation in bulbs of Lycoris.

Vegetative propagation (VP) is an important practice for production in many horticultural plants. Sugar supply constitutes the basis of VP in bulb flowers, but the underlying molecular basis remains elusive. By performing a combined sequencing technologies coupled with ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry approach for metabolic analyses, we compared two Lycoris species with contrasting regeneration rates: high-regeneration Lycoris sprengeri and low-regeneration Lycoris aurea. A comprehensive multi-omics analyses identified both expected processes involving carbohydrate metabolism and transcription factor networks, as well as the metabolic characteristics for each developmental stage. A higher abundance of the differentially expressed genes including those encoding ethylene responsive factors was detected at bulblet initiation stage compared to the late stage of bulblet development. High hexose-to-sucrose ratio correlated to bulblet formation across all the species examined, indicating its role in the VP process in Lycoris bulb. Importantly, a clear difference between cell wall invertase (CWIN)-catalyzed sucrose unloading in high-regeneration species and the sucrose synthase-catalyzed pathway in low-regeneration species was observed at the bulblet initiation stage, which was supported by findings from carboxyfluorescein tracing and quantitative real-time PCR analyses. Collectively, the findings indicate a sugar-mediated model of the regulation of VP in which high CWIN expression or activity may promote bulblet initiation via enhancing apoplasmic unloading of sucrose or sugar signals, whereas the subsequent high ratio of hexose-to-sucrose likely supports cell division characterized in the next phase of bulblet formation.

Rapid DNA interstrand cross-linking of Pt(IV) compound.

Pt(IV) anticancer compounds have been developed for several decades to overcome the drawbacks of their Pt(II) congeners, and the reduction of Pt(IV) to Pt(II) has been commonly regarded as a necessary step in the activation of Pt(IV) compounds prior to targeting DNA. However, blockage of glutathione (GSH) biosynthesis resulted in a slight effect on the cytotoxicity of oxoplatin in yeast Saccharomyces cerevisiae strains, urging us to reconsider the mechanism of actions for the "inert" Pt(IV) complexes. Using X-ray absorption near-edge spectroscopy (XANES), our data demonstrated that Pt(IV) complex oxoplatin could bind to DNA in a tetravalent state. Both alkaline denaturing agarose electrophoresis and thermal denaturation-renaturation assay revealed that oxoplatin could rapidly produce stable interstrand crosslinks (ICLs), which can further translate into a fast cell-killing process in cancer cells. Using quantitative real-time PCR and immunofluorescence analysis, we also proved that Pt(IV) complex oxoplatin could induce a quick intracellular response of the FA/BRCA pathway in cancer cells that involves the DNA interstrand crosslinking repair system, and this quick response to ICLs was independent with the intracellular GSH levels. Cell cycle analysis showed that short incubation with oxoplatin can induce a strong S phase arrest in HeLa cells, indicating that the rapid interstrand crosslinks produced by oxoplatin might stall the replication fork, result in the double-strand breaks, and eventually induce cell death. Our results implied that, besides the reduction mechanism to release the Pt(II) congeners, direct and rapid interstrand cross-linking with DNA by Pt(IV) compounds might be a unique mechanism for Pt(IV) compounds, which may provide new insight for the development of next-generation platinum-based drugs.

Silencing of circ_0000517 suppresses proliferation, glycolysis, and glutamine decomposition of non-small cell lung cancer by modulating miR-330-5p/YY1 signal pathway.

In recent years, circular RNA (circRNA) has been found to be involved in a variety of cancer processes. More and more attention has been paid to the research of circRNA in lung cancer. This study aims to investigate whether circ_0000517 affected the physiology of non-small cell lung cancer (NSCLC) and the underlying mechanism. The results demonstrated that circ_0000517 was highly expressed in lung cancer tissues and cells, and overexpression of circ_0000517 was negatively correlated with the prognosis of NSCLC patients. Silencing of circ_0000517 significantly inhibited the proliferation, glycolysis, and glutamine decomposition of NSCLC cells in vitro and repressed the growth of xenografted tumors in vivo. Moreover, knockdown of circ_0000517 attenuated the expression of PCNA, HK2, LDHA, ASCT2, and GLS1. Further study found that circ_0000517 targeted miR-330-5p and miR-330-5p targeted YY1. In addition, miR-330-5p inhibitor reversed inhibition of cancer cell proliferation, glycolysis, and glutamine decomposition induced by si-circ_0000517. In conclusion, our study revealed that silencing of circ_0000517 improved the progression of NSCLC through regulating miR-330-5p/YY1 axis.

[One-hour post-load plasma glucose: a better indicator of glucose metabolism for obstructive sleep apnea?]

OBJECTIVE: To assess the value of blood glucose at different time points in oral glucose tolerance test (OGTT), particularly one?hour post load plasma glucose (1 hPG), in evaluating glucose metabolism in adult patients with obstructive sleep apnea (OSA). METHODS: Eighty nine adultswith newly diagnosed OSA were analyzed retrospectively for sleep architecture assessed using polysomnography and glucose metabolism assessed by OGTT at different time points (0, 30, 60, 120, and 180 min). Pearson's correlatives and multiple linear regression models were established to investigate the correlations between glucose metabolism and other indices including sleep architecture, apnea hypopnea index (AHI), mean and lowest oxygen saturation (MSO2 and LSO2) and obesity measurements. RESULTS: The majority (67.4%) of the patients had abnormal 1 hPG, and 41.6% had abnormal 2 hPG. 1 hPG was positively correlated with neck circumference (r=0.245), abdomen circumference (r=0.231), systolic blood pressure (r=0.213), diastolic blood pressure (r=0.276) and AHI (r=0.324), and was negatively associated with MSO2 (r=-0.341) and LSO2 (r=-0.387) (all P<0.05). After controlling for age, BMI, neck and abdomen circumferences, 1 hPG was found to inversely correlated with MSO2 (r=-0.253, P=0.032) and LSO2 (r=-0.311, P=0.008). In non-obese OSA subgroup, 1 hPG was significantly associated with OSA-related indices, and regression models showed that LSO2 and N2 were the two most important contributors to 1 hPG (adjusted R2=0.349, P<0.001); plasma glucose at other time points did not show such correlations. CONCLUSIONS: 1 hPG abnormality occurs earlier than 2 hPG in OSA patients. 1 hPG is significantly associated with OSA independent of obesity and may serve as a better index for measuring OSA-related glucose disorder.

A Nomogram for Predicting the Likelihood of Obstructive Sleep Apnea to Reduce the Unnecessary Polysomnography Examinations.

BACKGROUND: The currently available polysomnography (PSG) equipments and operating personnel are facing increasing pressure, such situation may result in the problem that a large number of obstructive sleep apnea (OSA) patients cannot receive timely diagnosis and treatment, we sought to develop a nomogram quantifying the risk of OSA for a better decision of using PSG, based on the clinical syndromes and the demographic and anthropometric characteristics. METHODS: The nomogram was constructed through an ordinal logistic regression procedure. Predictive accuracy and performance characteristics were assessed with the area under the curve (AUC) of the receiver operating characteristics and calibration plots, respectively. Decision curve analyses were applied to assess the net benefit of the nomogram. RESULTS: Among the 401 patients, 73 (18.2%) were diagnosed and grouped as the none OSA (apnea-hypopnea index [AHI] <5), 67 (16.7%) the mild OSA (5 ≤ AHI < 15), 82 (20.4%) the moderate OSA (15 ≤ AHI < 30), and 179 (44.6%) the severe OSA (AHI ≥ 30). The multivariable analysis suggested the significant factors were duration of disease, smoking status, difficulty of falling asleep, lack of energy, and waist circumference. A nomogram was created for the prediction of OSA using these clinical parameters and was internally validated using bootstrapping method. The discrimination accuracies of the nomogram for any OSA, moderate-severe OSA, and severe OSA were 83.8%, 79.9%, and 80.5%, respectively, which indicated good calibration. Decision curve analysis showed that using nomogram could reduce the unnecessary polysomnography (PSG) by 10% without increasing the false negatives. CONCLUSIONS: The established clinical nomogram provides high accuracy in predicting the individual risk of OSA. This tool may help physicians better make decisions on PSG arrangement for the patients referred to sleep centers.

[Value of tonsillectomy in patient with obstructive sleep apnea hypopnea syndrome with Friedman staging I].

OBJECTIVE: To evaluate the availability of tonsillectomy in patients with obstructive sleep apnea hypopnea syndrome (OSAHS) staged as Friedman I. METHODS: Fifty-six patients with OSAHS in Friedman stage I who refused uvulopalatopharyngoplasty (UPPP) received tonsillectomy merely from January 2004 to March 2010. There were 20 mild, 24 moderate and 12 serious patients respectively in this group. The other 68 OSAHS patients in Friedman stage I received UPPP at the same time as matched group, including 26 mild, 28 moderate and 14 serious patients. RESULTS: There was no significant difference before operation in terms of age, body mass index, apnea hypopnea index (AHI), the lowest pulse oxygen saturation (SPO(2)) and average SPO(2) between the two groups. There were significant difference in mean length of operation (U = 0.000, P < 0.01), hospitalization day (U = 458.5, P < 0.01), visual analogue scale after surgery (U = 0.000, P < 0.01) in these two group. There was no significant difference in surgical effective rate between the two groups (χ(2) = 0.857, P > 0.05). There was also no significant difference in terms of age, body mass index, AHI, the lowest SPO(2) and average SPO(2) after operation between the two groups (t test P > 0.05). The surgical effective rate for the long term of the two groups was equal (χ(2) = 0.857, P > 0.05). Even patients with serious OSAHS in Friedman stage I, the surgical effective rate of the two groups was equivalent (Fisher's exact test, P > 0.05). CONCLUSIONS: Tonsillectomy is a safe and effective surgery for OSAHS in Friedman stage I, whose main structural load lies in the hypertrophic tonsil. It should be the first surgical choice for OSAHS in Friedman stage I.

BiClamp(®) forceps was significantly superior to conventional suture ligation in radical abdominal hysterectomy: a retrospective cohort study in 391 cases.

OBJECTIVE: To evaluate the efficacy and security of ERBE BiClamp(®) forceps in radical abdominal hysterectomy for managing those cervical cancers, extending to other gynecologic cancers such as endometrial cancer and ovarian cancer as well. METHODS: A retrospective cohort study was made in 391 cases from 450 FIGO IA2-IIB cervical cancers between November 2005 and September 2010. After baseline character analysis, the conventional group (n = 195) was compared with the BiClamp group (n = 196) on the basis of surgical outcome and complications. Data analysis was based on intention to treat with statistics software SPSS17.0. RESULTS: Comparison between conventional suture ligation and BiClamp(®) forceps is as follows: the operation time was 247.7 ± 47.7 min for the conventional suture ligation versus 224.1 ± 36.2 min (P < 0.001) for BiClamp(®) forceps, estimated blood loss was 769.2 ± 310.4 ml versus 534.8 ± 232.5 ml (P < 0.001), gauze consumption was 35.3 ± 10.6 sheets versus 28.2 ± 7.4 sheets (P < 0.001), intra-operative blood transfusion rate was 75.9 versus 28.1% (P < 0.001), hemoglobin decline was 29.2 ± 10.1 g/L versus 26.5 ± 9.2 g/L (P = 0.085), postoperative blood transfusion rate was 17.0 versus 15.6% (P = 0.818), closed suction drainage was 268.8 ± 162.0 ml versus 208.3 ± 141.7 ml (P < 0.001), hospital stay was 8.8 ± 2.5 days versus 7.1 ± 2.2 days (P < 0.001), postoperative complications was 23.6 versus 14.8% (P = 0.027). CONCLUSION: With obvious decrease of operation time, blood loss, postoperative complications, hospital stay and particularly, intra-operative blood transfusion rate, BiClamp(®) forceps has been proved more efficient and controllable in radical abdominal hysterectomies of cervical cancers than conventional suture ligations, extending to endometrial cancers and ovarian cancers, hence deserves to be popularized.

[Effects of proteasome inhibitor MG132 on cell proliferation, apoptosis, and cell cycle in HEC-1B and Ishikawa cells].

OBJECTIVE: To study the anti-cancer activities of Z-Leu-Leu-Leu-cho (MG132) against human endometrial carcinoma HEC-1B and Ishikawa cells and the potential of MG132 in human endometrial cancer therapy. METHODS: HEC-1B and Ishikawa cells were treated with MG132. Cell proliferation was assessed by MTT while cell apoptosis rate and cell-cycle distribution were assessed by flow cytometry. RESULTS: The proliferation of HEC-1B and Ishikawa cells was inhibited by MG132 and cell proliferation was significantly inhibited with the raised concentration of MG132 (P<0.01), Ishikawa cells were more sensitive than HEC-1B cells to MG132. MG132 could induce the apoptosis of HEC-1B and Ishikawa cells (P = 0.000). Cell cycle analysis indicated that the percentage of HEC-1B cells was increased in G2 phase (P<0.05) while Ishikawa cells' was increased in G1 and G2 phase (P<0.05). CONCLUSION: Proteasome inhibitor MG132 could inhibit the proliferation, promote cell apoptosis, and block the cell cycle of HEC-1B and Ishikawa cells. MG132 may be a potential treatment for endometrial cancer.

[Immune function changes in patients with obstructive sleep apnea hypopnea syndrome].

OBJECTIVE: To explore the potential changes in the immune function of patients with obstructive sleep apnea hypopnea syndrome (OSAHS). METHODS: We carried out a retrospective cross-sectional study of 187 patients with established OSAHS and 20 healthy subjects (control). For all the patients, the medical history was carefully examined, and overnight sleep monitoring was carried out with detection of the humoral and cellular immunity. RESULTS: We found a significant increase in the levels of C3 and a decrease in both the IgM level and NK cell percentage in OSAHS patients as compared to the control group (P<0.01). Correlation analysis indicated that C3 was positive correlated to AHI but inversely to the lowest pulse oxygen saturation (LSpO(2)); IgM showed a mild positively correlation to LSpO(2), and NK cells had a mild inverse correlation to AHI. The other immunological indices were not found to undergo noticeable changes or show correlations in OSAHS. CONCLUSION: Immune function changes occur in patients with OSAHS, characterized primarily by deteriorations in the humoral and cellular immunity.

Oral supplementation of catalpol ameliorates diabetic encephalopathy in rats.

Diabetes mellitus can cause dysfunction of the central nervous system called "diabetic encephalopathy." Although insulin and various oral drugs are used to treat diabetes, they do not completely prevent the development of diabetic encephalopathy, and novel strategies for the prevention and treatment are urgently needed. Catalpol, an iridoid glycoside, has properties of anti-inflammation, antioxidant and decreasing blood glucose level and thus has the possibility of treating diabetic encephalopathy. Therefore, the study was designed to investigate the effects of catalpol on diabetic encephalopathy in rats. A single dose of 65 mg/kg streptozotocin was injected intraperitoneally to induce diabetes. Intragastric infusion of catalpol was performed for 6 weeks with the doses of 10, 50 and 100 mg/kg, respectively. The Y-type maze test, biochemical measurement, Nissl staining and the terminal deoxynucleotidyl transferase-mediated UTP nick end labeling methods were used to evaluate the neuropathological changes and the effects of catalpol on diabetic rats. The results showed that streptozotocin-induced diabetes produced obvious neuron damage and cognitive dysfunction coupling with markedly increased oxidative stress in the brain. Long-term oral supplementation of catalpol improved neuronal injury and cognitive dysfunction by attenuating oxidative stress. The effects that catalpol could both increase the nerve growth factor concentration and decrease the blood glucose level were related with the function of defending against oxidative stress of catalpol. The study suggested that oral supplementation of catalpol might be a potential therapeutic strategy for the treatment and/or prevention of diabetic encephalopathy.

Catalpol modulates the expressions of Bcl-2 and Bax and attenuates apoptosis in gerbils after ischemic injury.

Our previous study described the neuroprotective effects of catalpol in gerbil ischemic model, in which catalpol was shown to prevent hippocampal neurons from death and ameliorate the cognitive ability of the animals. In the study, we focused on investigating the neuroprotective mechanism of catalpol. Animals were randomly assigned three groups as sham-operated, ischemia-treated with saline and ischemia-treated with catalpol. Transient global ischemia was produced by a 5 min occlusion of the bilateral common carotid arteries. Catalpol was intraperitoneally injected at the dose of 5 mg/kg immediately after reperfusion and repeatedly at 12, 24, 48 and 72 h. Histology as well as immunohistochemistry and TUNEL (the terminal deoxynucleotidyl transferase-mediated UTP nick end label) analysis were performed on serial slices through the dorsal hippocampus after gerbils were sacrificed. The results showed that 5 min transient global ischemia followed by 4 days reperfusion caused significant increases in TUNEL-positive and Bax-positive cells in hippocampal CA1 subfield. Catalpol not only significantly reduced TUNEL-positive and Bax-positive cells but also significantly increased Bcl-2-positive cells. All these suggested that catalpol could effectively inhibit apoptosis by modulating the expressions of Bcl-2 and Bax genes.

Catalpol prevents the loss of CA1 hippocampal neurons and reduces working errors in gerbils after ischemia-reperfusion injury.

Catalpol, an iridoid glycoside, contained richly in the roots of Rehmannia glutinosa, was found for the first time to be of neuroprotection in gerbils subjected to transient global cerebral ischemia. Catalpol (1 mg/kg ip) used immediately after reperfusion and repeatedly at 12, 24, 48 and 72 h significantly rescued neurons in hippocampal CA1 subfield and reduced working errors during behavioral testing. The neuroprotective efficacy of catalpol became more evident when the doses of catalpol were increased to 5 and 10mg/kg. In addition, it was exciting that the significant neuroprotection by catalpol was also evident when catalpol was applied up to 3 h after ischemia. But the neuroprotective efficacy of catalpol became weak when catalpol was given at 6h after ischemia. Of great encouragement was the finding that the neuroprotection of catalpol could be seen not only in a short post-ischemic period (12 days) but also in a long period (35 days). All these indicated that catalpol was truly neuroprotective rather than simply delayed the onset of neuronal damage and might be of therapeutic value for the treatment of global cerebral ischemia.

Neuroprotective properties of catalpol in transient global cerebral ischemia in gerbils: dose-response, therapeutic time-window and long-term efficacy.

The present study evaluated for the first time the dose-effectiveness, therapeutic time-window and long-term efficacy of the neuroprotection of catalpol by behavioral and histological measures in gerbils subjected to transient global cerebral ischemia. Catalpol (1 mg/kg ip) used immediately after reperfusion and repeatedly at 12, 24, 48 and 72 h significantly rescued neurons in the hippocampal CA1 subfield and reduced cognitive impairment. The neuroprotective efficacy of catalpol became more evident at the doses of 5 and 10 mg/kg. Of great importance were the findings that the neuroprotective efficacy of catalpol still could be seen even when the treatment was delayed 3 h and when the observational period was lasted out 35 days after ischemia. It was reasonable to draw the conclusion that catalpol was truly neuroprotective rather than simply delayed the onset of neuronal damage. These results suggested that catalpol might be of therapeutic value for global cerebral ischemia.

Neuroprotection of catalpol in transient global ischemia in gerbils.

The neuroprotection of catalpol and its mechanism was evaluated in cerebral ischemic model in gerbils. Three groups were designed as sham-operated, ischemia-treated, respectively, with catalpol and saline. Catalpol was injected intraperitoneally immediately after reperfusion and repeatedly at 12, 24, 48 and 72 h with the dose of 5.0 mg/kg. The neuroprotection was estimated by the indexes of behavior and histology. Behavioral testing was performed in Y-maze and the survival neurons in CA1 subfield were counted under a microscope after behavioral testing. In addition, apoptosis induced by ischemia was also examined by using the terminal deoxynucleotidyl transferase-mediated UTP nick end labeling method. It was shown that catalpol significantly attenuated apoptosis, rescued hippocampal CA1 neurons and reduced cognitive impairment. In order to make clear the mechanism of catalpol's neuroprotection, the activities of endogenous antioxidants and nitric oxide synthase together with the content of lipid peroxide in cortex and hippocampus were assayed. The results proved that catalpol significantly reduced the content of lipid peroxide, increased the activity of glutathione peroxidase and decreased the activity of nitric oxide synthase. All these suggested that catalpol was a potential neuroprotective agent and its neuroprotective effects were achieved at least partly by promoting endogenous antioxidant enzymatic activities and reducing the formation of nitric oxide.

[Analysis of genetic effects for growth traits of Eucalyptus globulus Labill. in a 6 x 6 diallel design].

The genetic effects of general and specific combining ability, male, female, and specific reciprocal cross were estimated for height, diameter of breast height (dbh), volume, crown-width, under branch height, crook degree and ratio of branch with shoot in Eucalyptus globulus Labill. Three quantitative characters, i.e. tree height, dbh, volume were focused, and their effects were discussed in detail. It was found that the GCA, SCA and specific reciprocal cross effects (SRE) were most important, and female effect (FEM) and male effect (MAL) were less important. Three parents with high GCA were good materials used for establishing seed orchard. Eight combinations significantly improved quantitative characters. One of them was improved on quantitative character and trunk quality at the same time. The breeding strategy and methods were also suggested for E. globulus.