Effect of miR-202-5p-mediated ATG7 on autophagy and apoptosis of degenerative nucleus pulposus cells.

OBJECTIVE: This study aimed to research the effect of miR-202-5p-mediated ATG7 on autophagy and apoptosis of degenerative nucleus pulposus cells. PATIENTS AND METHODS: The intervertebral disc nucleus pulposus (NP) tissue of patients with intervertebral disc degenerative disease and normal intervertebral disc nucleus pulposus (NP) tissue of patients with spinal fractures was collected as the research object. Normal NP cells and degenerative NP cells were isolated. Low expression of miR-202-5p and overexpression of ATG7 were carried out in degenerative NP cells. The expression of miR-202-5p and ATG7 mRNA was detected by RT-PCR. The expression of ATG7, LC3-II, Bax, and Bcl-2 proteins was detected by Western blot. The autophagy of cells was detected by MDC staining. The apoptosis of NP cells was detected by flow cytometry. The targeting relationship between miR-202-5p and ATG7 was detected by Dual-Luciferase reporter. RESULTS: In the degenerative NP tissues, miR-202-5p was highly expressed and ATG7 was low expressed. The inhibition of miR-202-5p expression can effectively promote autophagy of NP cells, increase the expression of ATG7 and LC3-II, inhibit the apoptosis of NP cells, inhibit the expression of pro-apoptotic proteins Bax, and promote the expression of pro-apoptotic proteins Bcl-2 proteins. The upregulation of ATG7 expression in degenerative NP cells alone had the same effect as the downregulation of miR-202-5p. The assay of the Dual-Luciferase reporter confirmed the targeting relationship between miR-202-5p and ATG7. CONCLUSIONS: MiR-202-5p can affect the autophagy and apoptosis of degenerative nucleus pulposus cells through targeted adjustment of ATG7, which may be a new therapeutic target for intervertebral disc degenerative diseases.

Beinaglutide showed significant weight-loss benefit and effective glycaemic control for the treatment of type 2 diabetes in a real-world setting: a 3-month, multicentre, observational, retrospective, open-label study.

AIMS: The purpose of this study was to examine the effectiveness of beinaglutide on body weight, glycated haemoglobin (HbA1c), blood pressure and lipid profiles in patients with type 2 diabetes mellitus (T2DM) in a real-world setting in China. MATERIALS AND METHODS: This was a multicentre, observational, retrospective, open-label study conducted in China. Data were collected from T2DM patients who started treatment with beinaglutide between 2017 and 2018. RESULTS: A total of 314 patients were included in the study. After 3 months of treatment with beinaglutide, there were significant reductions in body weight (-10.05 kg [95% confidence interval -9.29 to -10.80]), HbA1c (-2.87% [-2.62 to -3.11]), 2-h postprandial plasma glucose (-5.46 mmol L-1 [-4.96 to -5.95]) and fasting plasma glucose (-3.04 mmol L-1 [-2.78 to -3.31]) (all p < 0.0001). In addition, 84.96% and 72.18% of the patients achieved weight loss of ≥5% and ≥10%, respectively. Subgroup analyses showed that weight loss was significantly greater in patients with ≥28 kg m-2 of baseline body mass index and 0.60 mg of beinaglutide doses (p = 0.007 and p < 0.0001, respectively). HbA1c reductions were significantly greater in patients with ≥9.0% baseline HbA1c and in those administered 0.40-0.48 mg of beinaglutide doses (all p < 0.0001). Weight loss at 3 months was positively correlated with baseline BMI and the dose of beinaglutide. Positive determinants for HbA1c reduction after 3 months were baseline HbA1c and the dose of beinaglutide. CONCLUSIONS: These observational results confirmed the benefits of beinaglutide in weight loss and glycaemic control and support the use of beinaglutide as an effective treatment for T2DM.

MicroRNA-449b-5p promotes the progression of osteoporosis by inhibiting osteogenic differentiation of BMSCs via targeting Satb2.

OBJECTIVE: We aimed to explore the role of microRNA-449b-5p in osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) and its mechanism of action. MATERIALS AND METHODS: Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay was used to detect the expression levels of microRNA-449b-5p and osteogenic markers including RUNX2, OCN during BMSCs differentiation. The microRNA-449b-5p mimic and microRNA-449b-5p inhibitors were transfected into BMSCs to achieve microRNA-449b-5p overexpression and knockdown, then the expressions of osteogenic markers were detected by qRT-PCR. The ALP activity staining and the alizarin red staining were used to detect the activity of ALP and the mineralization ability of cells after overexpression and knockdown of microRNA-449b-5p. Binding sites for microRNA-449b-5p and Satb2 were predicted by TargetScan, the PicTar and microRNAanda programs, and confirmed by dual-luciferase reporter gene assay. The relationship between microRNA-449b-5p and Satb2 was analyzed by QRT-PCR and Western blot. The microRNA-449b-5p inhibitor and shSATB2 lentivirus were simultaneously transfected in BMSCs, and the expression levels of RUNX2, OCN and ALP were detected by qRT-PCR and ALP activity assays. RESULTS: microRNA-449b-5p expression gradually decreased during osteogenic differentiation. Overexpression of microRNA-449b-5p inhibited BMSCs differentiation by down-regulating ALP activity, RUNX2, and OCN expression, while the opposite result was observed after knockdown of microRNA-449b-5p. MicroRNA-449b-5p can bind to the 3'UTR end of Satb2, which was involved in the osteogenic differentiation of microRNA-449b-5p-regulated BMSCs, and silencing of Satb2 can abolish the positive effect of the microRNA-449b-5p inhibitor on osteoblasts differentiation. CONCLUSIONS: microRNA-449b-5p could aggravate osteoporosis by inhibiting osteogenic differentiation of BMSCs through targeting Satb2.

[Prognosis analysis of primary breast diffuse large B cell lymphoma].

Objective: To investigate the clinical characteristics, therapy modality and prognosis of primary breast diffuse large B-cell lymphoma(PB-DLBCL). Methods: A total of 68 patients with PB-DLBCL treated in Tianjin Medical University Cancer Institute and Hospital were enrolled between January 1, 2004 and January 31, 2017. Clinicopathological data were retrospectively analyzed. 67 patients were female and only one male. The median age was 56 years old. 46 patients had Ann Arbor clinical stageⅠ~Ⅱ disease, and the other 22 were stage Ⅲ~Ⅳ. The patients with and without B symptom were 11 and 57, respectively. Kaplan-Meier method was used for univariate analysis to calculate the 5-year overall survival (OS) rate and 5-year progress-free survival (PFS) rate, compared using the log rank test. Cox regression analysis was used for multivariate analysis. Results: The 1, 3, 5-year OS rate were 84.0%, 78.0% and 73.0%, and 1, 3, 5-year PFS rate were 80.0%, 71.0% and 51.0%, respectively. Univariate analysis indicated that eastern cooperative oncology group (ECOG) score, Ann Arbor clinical stage, international prognostic index (IPI) score, risk stratification, B symptom, ß2-microglobulin(ß2-MG) level, size of the tumor and cycles of chemotherapy were prognostic factors for OS (all P<0.05), and Ann Arbor clinical stage, IPI score, risk stratification and B symptom were prognostic factors for PFS (all P<0.05). Multivariate analysis indicated that Ann Arbor clinical stage was independent prognostic factor for OS(P=0.029) and B symptom was independent prognostic factor for PFS(P=0.028). Conclusions: Prognosis of PB-DLBCL was relatively good. Ann Arbor clinical stage and B symptom were independent prognostic factors for OS and PFS, respectively.

[Experimental study of using chitosan nerve conduit combined with PRP to repair facial nerve defect].

Objective:The purpose of this study was to produce chitosan nerve conduit in order to investigate the feasibility of chitosan nerve conduit combined with autologous platelet-rich plasma (PRP) for repairing facial nerve defects.Method:Forty New Zealand white rabbits were randomly divided into four groups (n=10 in each group).Establishment of the facial nerve defect model of the upper buccal branches was placed in the nerve regeneration catheter and injected with the same amount of PRP and saline. The PRP was injected into the chitosan nerve conduit as group A. The physiological saline was injected into the chitosan nerve conduit as group B. The physiological saline was injected into the silicone tube as group C. The PRP was injected into the silicone tube as group D. Eight weeks later,facial nerve gross observation,facial nerve electrophysiological tests,histological observation,image analysis,valuation of nerve regeneration recovery were detected.Result:Five patients were lost to follow up (all five in the modified Semont group),and three patients failed to complete treatment (all three in the Epley group). The sequelae at the 3rd day and one week after modified Semont maneuver were 27 and 9,while 41 and 15 in Epley group. The efficacy rates at the 3rd day and one week after modified Semont maneuver were 91.7% and 98.3%,and 91.9% and 96.8% in Epley group retrospectively. The sequelae and short-term effective rate of patients in modified Semont group was no difference when compared with that in Epley group (P>0.05).Conclusion:The chitosan nerve conduit combined with PRP has a certain effect on the repair of facial nerve defects and is expected to be applied to the repair of clinical facial nerve defects.

Thermotherapy-induced reduction in glioma invasiveness is mediated by tumor necrosis factor-alpha.

Thermotherapy has been proven to be effective for the treatment of various tumors, including glioma. We determined whether tumor necrosis factor-alpha (TNF-α) is involved in the regulation of the biological processes of glioma development. Reverse transcription-polymerase chain reaction (RT-PCR) and immunocytochemistry were used to investigate the levels of TNF-α mRNA and heat shock factor-1 (HSF1) protein, respectively, in glioma cells. Radioimmunoassay was used to dynamically monitor the contents of TNF-α in the nutrient fluid of C6 cells after thermotherapy treatment. Crystal violet staining was used to determine glioma invasiveness. The most obvious increases in HSF1 protein and TNF-α mRNA in C6 cells were observed at 30 and 60 min after thermotherapy, respectively. In addition, the radioactivity of TNF-α in the culture fluid of the C6 cells reached a peak after 120 min of thermotherapy. In addition, glioma invasiveness decreased and the concentration of TNF-α reached a maximum after 120 min of thermotherapy. Our results show that the decrease in thermotherapy-mediated glioma invasiveness is due to the accelerated release of TNF-α, which could promote the release of HSF1 from neurospongioma cells.

Effect of delivery room temperature on the admission temperature of premature infants: a randomized controlled trial.

OBJECTIVE: To determine if increasing delivery room temperature to that recommended by the World Health Organization results in increased admission temperatures of preterm infants. STUDY DESIGN: Admission rectal temperatures of newborns ≤32 weeks gestation delivered in rooms with temperature set at 24 to 26 °C were compared with those of similar newborns delivered in rooms with temperature set at 20 to 23 °C. RESULT: Premature newborns delivered in rooms with mean temperature 25.1±0.6 °C (n=43), compared with those delivered in rooms with mean temperature 22.5±0.6 °C (n=48), had a lower incidence (34.9% vs 68.8%, P<0.01) of admission rectal temperature <36 °C and higher admission rectal temperatures (36.0±0.9 °C vs 35.5±0.8 °C, P<0.01). This difference persisted after adjustment for birth weight and 5 min Apgar score. CONCLUSION: Increasing delivery room temperatures to that recommended by the World Health Organization decreases cold stress in premature newborns.

[A pharmacological study on suppository for anti-prostatis].

OBJECTIVE: To approach the pharmacological mechanism of the suppository for anti-prostatis (SAP). METHOD: Investigating the inflammation of prostate in rat models with bacterial and nonbacterial prostatis induced by colonbacillus and "Xiao Zhi Ling", as well as the change of rheology in the rat model with acute blood stasis. Several tests on anti-inflammation, analgesia and bacteriostasis were conducted. RESULT AND CONCLUSION: SAP could restrain the inflammation of prostate and resume its secretion obviously. It is very effective for anti-inflammation, analgesia and bacteriostasis, and also helps improve the change of theology in rats with blood stasis.

Selective modulation of excitatory transmission by mu-opioid receptor activation in rat supraoptic neurons.

Opioid peptides have profound inhibitory effects on the production of oxytocin and vasopressin, but their direct effects on magnocellular neuroendocrine neurons appear to be relatively weak. We tested whether a presynaptic mechanism is involved in this inhibition. The effects of mu-opioid receptor agonist D-Ala(2), N-CH(3)-Phe(4), Gly(5)-ol-enkephalin (DAGO) on excitatory and inhibitory transmission were studied in supraoptic nucleus (SON) neurons from rat hypothalamic slices using whole cell recording. DAGO reduced the amplitude of evoked glutamatergic excitatory postsynaptic currents (EPSCs) in a dose-dependent manner. In the presence of tetrodotoxin (TTX) to block spike activity, DAGO also reduced the frequency of spontaneous miniature EPSCs without altering their amplitude distribution, rising time, or decaying time constant. The above effects of DAGO were reversed by wash out, or by addition of opioid receptor antagonist naloxone or selective mu-antagonist Cys(2)-Tyr(3)-Orn(5)-Pen(7)-NH(2) (CTOP). In contrast, DAGO had no significant effect on the evoked and spontaneous miniature GABAergic inhibitory postsynaptic currents (IPSCs) in most SON neurons. A direct membrane hyperpolarization of SON neurons was not detected in the presence of DAGO. These results indicate that mu-opioid receptor activation selectively inhibits excitatory activity in SON neurons via a presynaptic mechanism.

[Whole-cell recordings of the supraoptic nucleus neurons from rat hypothalamic slices in vitro].

Fifty-two supraoptic nucleus neurons in rat slice preparations were studied using whole cell patch-clamp technique. The mean passive and active membrane properties were measured as follows: resting membrane potential, 59 +/- 8 mV; input resistance, 535 +/- 129 M omega; time constant, 32 +/- 9 ms; amplitude of the action potentials, 99 +/- 11 mV; overshoot, 37 +/- 13 mV (n = 39). Most of these neurons showed a prominent slow after-hyperpolarization potential or current in response to depolarizing pulse. In votage-clamp condition, it was found that virtually all supraoptic neurons (n = 13) were invaded by spontaneous synaptic inputs. Pharmacological experiments showed that the excitatory postsynaptic currents (EPSCs) were mediated by non-NMDA glutamate receptors, whereas inhibitory postsynaptic currents (IPSCs) by GABAA receptors.

Nitric oxide inhibits neuronal activity in the supraoptic nucleus of the rat hypothalamic slices.

The presence of abundant nitric oxide synthase (NOS) in magnocellular neurons of the rat hypothalamus suggests that nitric oxide (NO) may be involved in controlling the release of oxytocin and vasopressin. To test this possibility, we examined the effect of NO-related drugs on extracellular discharges of 124 supraoptic nucleus (SON) neurons from slices of rat hypothalamus in vitro. Twenty-three (43%) of 53 neurons were inhibited by sodium nitroprusside (SNP), a spontaneous releaser of NO, at 1-3 mM. This inhibition was prevented by preincubation of the slices with 1 microM hemoglobin, an inactivator of NO (n = 14), whereas hemoglobin alone enhanced neuronal activity in seven (35%) of 20 neurons. L-Arginine (1 mM), a precursor of NO, inhibited neuronal activity in five (36%) of 14 neurons, while D-arginine (1 mM), the inactive counterpart of L-arginine, was ineffective (n = 12). N-omega-nitro-L-arginine methyl ester (L-NAME, 10 microM), an inhibitor of NOS, also enhanced neuronal activity in five (29%) of 17 neurons, while N-omega-nitro-D-arginine methyl ester (DNAME, 10 microM), the inactive enantiomer of L-NAME, was without effect (n = 11). Together, our data show that NO exerts predominantly an inhibitory effect on SON neurons and may serve as a negative feedback loop in controlling release of oxytocin and vasopressin.

Nitric oxide synthase expression in vagal complex following vagotomy in the rat.

Peripheral axotomy of the spinal nerve and avulsion of the ventral roots have been found to induce increase in expression of nitric oxide synthase (NOS) in the spinal motor neurones and the dorsal root ganglion. The present study investigated changes of NOS, using NADPH-diaphorase (NADPH-d) reactivity as the marker, in vagal complex after axotomy in the rat. Eight days after left cervical vagotomy the NADPH-d reactivity was found to be markedly enhanced in the dorsal motor nucleus of the vagus nerve, the ambiguus nucleus, the solitary tract and the nucleus of the tractus solitarius, and the nodose ganglion. This study offers the first evidence of changes in NOS expression in cranial visceral components following axotomy.