Air Pollutants, Occupational ; analysis ; Solvents ; analysis

OJECTIVETo explore whether different acupuncture signals were afferent to the cerebellar fastigial nucleus (FN) neuron and to find out their corresponding effect features through observing the effect of spontaneous discharge of cerebellar FN neuron by needling at different acupoints.

METHODSTotally 120 male SD rats were anesthetized by 20% urethane and their right cerebellar FN were positioned (AP 11. 6 mm, RL 1. 0 mm, H 5. 6 mm). Extracelluar discharge was recorded by glass microelectrode (AP: -11. 6 mm, R: 1. 0 mm, H: 5.7 -7. 0 mm), using extracellular microelectrode recording method, recording the spontaneous discharge of cerebellar FN neurons as a baseline. Random order of needling at zusanli (ST36), quchi (Lil1), weishu (BL21), and zhongwan (CV12) were compared with the baseline before each acupuncture. Their effects on the discharge of cerebellar FN neurons were observed and compared with baselines.

RESULTSThe frequency of FN neuronal discharge could be elevated by needling at zusanli (ST36), quchi (LiI), weishu (BL21), and zhongwan (CV12) (P <0. 01, P <0. 05). The response rate of needling at Zhongwan (CV12, 56. 00%) was higher than that of needling at Zusanli (ST36), Quchi (Ll1), and Weishu (BL21) (35. 00%, 34. 62%, 36. 63%, respectively) with statistical difference (P <0. 05). The response rate of needling at zhongwan (CV12) was obviously higher than that of needing at other points (F = 2. 101, P < 0. 05).

CONCLUSIONSNeedling at zusanli (ST36 ), quchi (Lil), weishu (BL21), and zhongwan (CV12) could elevate the spontaneous discharge frequency of cerebellar FN neurons. Needling at Zhongwan (CV12) had advantageous roles in regulating cerebellar FN.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Cerebellar Nuclei ; physiology ; Male ; Microelectrodes ; Neurons ; Rats ; Rats, Sprague-Dawley

Objective To determine the effect of knocking down zebrafish faf1 gene by CRISPR/Cas9 editing technique.Methods gRNA was designed and prepared for the faf1 gene of zebrafish,and gRNA was mixed with Cas9 mRNA by microinjection into zebrafish single cell embryos.The mutant F0 generation zebrafish was screened out by enzyme digestion and gene sequencing.The mutant F0 was genetically outcrossed with the wild-type zebrafish to get the F1 heterozygous zebrafish,and the genotype of zebrafish was detected by microscopic observation.Results The faf1 gRNA and Cas9 mRNA were successfully prepared.The gRNA (gRNA6) located in the exon 6 of faf1 could shift the faf1 gene into frameshift mutations.The mutation type MU1 was screened out and the somatic cytochrome deposition delay was observed in this heterozygous zebrafish.At 4 d post fertilization (dpf),there were sarcomeric dysplasia and head shrinkage,increased hyoid angle and other craniofacial cartilage deformities.And the zebrafish died at 8 ～9 dpf.Conclusion CRISPR/Cas9 knocking out thefaf1 gene produces a new phenotype for zebrafish,with delayed pigment deposition and nodule-like change in tail muscle section.

OBJECTIVEEndothelial apoptosis plays an important role in the initiation of atherosclerosis. It would be useful to clarify whether activation of non-neuronal muscarinic receptor (NNMR) could prevent endothelial apoptosis and atherosclerosis. We investigated the effects of NNMR activation on regulating rat aortic endothelial cells (RAECs) apoptosis induced by homocysteine, an independent risk factor of atherosclerosis, and further studied its molecular mechanism.

METHODSRAECs were incubated using homocysteine at the concentration of 2.7 mmol/L for 36 h. RAECs were also pre-treated with carbachol or arecoline to examine their effects. RT-PCR was used to assess changes in the gene expression related to cell apoptosis.

RESULTSIncubation of RAECs with homocysteine at the concentration of 2.7 mmol/L resulted in morphologic changes, such as cellular shrinkage, membrane blebbing, chromatin condensation and margination. These could be attenuated by pretreatment with carbachol and arecoline at the concentration of 10 micromol/L for 12 h. Homocysteine induced apoptosis in RAECs and the molecular mechanisms were associated with the regulation of fas, fas-L and caspase-8 in the death receptor pathway, bcl-2, bcl-xL and bax in the mitochondrial pathway, caspase-12 in the endoplasmic reticulum pathway and caspase-3, caspase-6 and p53 as downstream effectors. Carbachol and arecoline attenuated the effects of homocysteine on genes in the death receptor pathway, in the mitochondrial pathway and in the downstream pathway. Atropine could reverse all of the effects of arecoline.

CONCLUSIONActivation of NNMR by carbacol and arecoline inhibits homocysteine-induced endothelial cell apoptosis mainly through regulation of death receptor pathway, mitochondrial pathway and downstream effectors.

Animals ; Aorta ; cytology ; Apoptosis ; Apoptosis Regulatory Proteins ; metabolism ; Arecoline ; Carbachol ; Cell Cycle ; Endoplasmic Reticulum ; metabolism ; Endothelial Cells ; cytology ; drug effects ; Homocysteine ; adverse effects ; Mitochondria ; metabolism ; Rats ; Receptors, Muscarinic ; metabolism

Objective To investigate the presence of mesenchymal stem cells ( MSC) in laryngeal mucosa, and to seek for the method to isolate, cultivate and identify these cells. Methods Normal laryngeal mucosa was obtained from patients with laryngeal carcinoma during surgery, and the generating mesenchymal cells was obtained by digestive method. The cell growth curve was evaluated by 3-(4. 5-methylthiozol-2yl)-2. 5-diphenyltetrazolium bromide ( MTT) assay. Colony forming cell assay was used to screen different morphologic colonies and evaluate clone formation ability. Flow cytometry was performed for the expression of the cells of the 4th passage surface marker profiles. Multiple differentiation potentials were confirmed by adipogenic, osteogenic and neural lineages induction. Results MTT assay and colony forming cell assay showed that laryngeal mucosa MSC had a relatively rapid proliferation capacity and a relatively high clone formation capacity ( clone formation rate 7. 1 % ). Flow cytometry analysis revealed that the laryngeal mucosa MSC were positive for CD29 (16. 9% ), CD44 (97.4% ), CD90 ( 89. 5% ), CD105 ( 85. 9% ), CD146 (2. 5% ) and stro-1 ( 20.4% ), but negative for CD34 (1.2%) and CD45 (0. 8% ). Laryngeal mucosa MSC undergone adipogenic, osteogenic and neural lineages induction were positive for oil red staining, alizarin red staining and S100 staining respectively, which suggested that laryngeal mucosa MSC could differentiate into adipogenic, osteogenic and neural lineages. Conclusion This study demonstrated that MSC with rapid proliferative capacity and multiple differentiation potential could be obtained from lamina propria of laryngeal mucosa.

OBJECTIVETo explore the feasibility of inducing human umbilical cord mesenchymal stem cells (HuMSCs) to differentiate into Leydig cells through conditioned medium derived from Leydig cells.

METHODSHuMSCs and Leydig cells were obtained by tissue blocks culture attachment and enzymatic digestion respectively. HuMSCs were induced by conditioned medium of Leydig cells as an experiment group while those before induction were cultured as a control group. The expressions of LHR, 3β-HSD and StAR in the induced HuMSCs were determined by RT-PCR after 3, 7 and 10 days of culture; those of CYP11A1, CYP17A1 and 3β-HSD measured by immunofluorescence staining after 2 weeks; and that of 3β-HSD detected by Western blot after 4 weeks.

RESULTSThe experimental group showed positively expressed LHR, 3β-HSD and StAR at 3, 7 and 10 days, CYP11A1, CYP17A1 and 3β-HSD at 2 weeks, and 3β-HSD at 4 weeks, while the control group revealed negative expressions at all the time points.

CONCLUSIONInduced with conditioned culture medium derived from Leydig cells, HuMSCs are likely to differentiate into steroidogenic cells and eventually into Leydig cells.

Cell Differentiation ; Culture Media, Conditioned ; Humans ; Leydig Cells ; cytology ; Male ; Mesenchymal Stromal Cells ; cytology ; Umbilical Cord ; cytology

Adult ; Humans ; Lipomatosis, Multiple Symmetrical ; Male ; Middle Aged

The antitussive activity assay for the root extraction of Disporum cantoniense was carried out with coughing mice induced by ammonia liquor. The results showed that the ethanol and water extractions of D. cantoniense possess strong antitussive activity, and the high dose of the former was better than positive control, and then the constituents of the ethanol extraction were separated and purified by various modern chromatographic techniques. Their structures were identified by physico-chemical properties and spectroscopic data. As a result, eight compounds were isolated and identified as stigmast-4-en-3-one(1), (22E, 24R)-ergosta-5, 7, 22-trien-3beta-ol(2), obtucarbamate A(3), obtucarbamate B(4), neotigogenin(5), azo-2, 2'-bis[Z-(2,3-dihydroxy-4-methyl-5-methoxy) phenyl ethylene] (6),dimethyl {[carbonylbis (azanediyl)] bis( 2-methyl-5, 1-phenylene) j dicarbamate (7) , and quercetin-3-O-pB-D-glucopyranoside(8). All compounds were isolated from this plant for the first time, and the result of bioactivity-directed isolation showed that compounds 3, 4, and 6 had obvious effect on antitussive activity, and compound 6 had the same level as positive control.
Animals ; Antitussive Agents ; chemistry ; isolation & purification ; pharmacology ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; pharmacology ; Ethanol ; chemistry ; Female ; Liliaceae ; chemistry ; Male ; Mice

Objective To observe the effect of low dosage of uhrashortwave(USW) on infarction volume, B cell lymphocytoma-xl (Bcl-xl) and tumor necrosis factor alpha (TNF-?) after cerebral ischemia-reperfusion in rats and discuss its acting mechanisms. Methods Focal ischemia-reperfusion model was established in 25 rats by re- versible right middle cerebral artery occlusion with filament. The right side cerebral ischemia was lasted for 2 hours and then followed with 24 hours of reperfusion. The content of neurological deficits were evaluated by the Zea-Longa 5-degree scoring system to select rats. After surgery, the rats were divided into 3 groups: blank control group, control group and USW treatment group. The brain of all rats was taken at 24 hours after reperfusion. The cerebral infarction volume, the expression of Bcl-xl and TNF-?were measured and analyzed. Results Twenty-five rats were used in the analysis of results. When compared with the control group, the infarction volume and rate in total cerebral volume of USW group significantly decreased (t = 2.54, 2.33, P

BACKGROUND:Laminoplasty and laminectomy are the most commonly used treatment methods for multilevel cervical spondylotic myelopathy, which is more common in elderly patients. C5 nerve root palsy is the common postoprative complication after posterior cervical repair. OBJECTIVE:To compare the incidence of C5 nerve root palsy after laminoplasty with mini-titanium plate fixation and laminectomy with internal fixation in repair of multilevel cervical spondylotic myelopathy. METHODS:Total y 134 patients with multilevel cervical spondylotic myelopathy from August 2010 to December 2014 were enrol ed, and then divided into laminoplasty group (n=45) and laminectomy group (n=89) owing to different ways of repair. Patients in the laminoplasty group were treated with laminoplasty with mini-titanium plate fixation, and patients in the laminectomy group were treated with laminectomy with internal fixation. C5 nerve root palsy condition after repair was recorded and evaluated. The cervical lordosis angle (Cobb angle) and cervical curvature index were compared. The Japanese Orthopaedic Association score was used for neurological assessment. RESULTS AND CONCLUSION:Al patients in both groups were fol owed up for more than 6 months. There were no significant differences in cervical lordotic angle and cervical curvature index at the first week before and after the treatment between these two groups (P>0.05). The Japanese Orthopaedic Association scores of patients after 6 months of treatment were significantly improved compared with that before treatment in these two groups (P<0.05). There were 2 cases of C5 nerve root palsy after the treatment of laminoplasty, the occurrence rate was 4%(2/45);there were 10 cases of C5 nerve root palsy after the treatment of laminectomy, the occurrence rate was 11%(10/89);there was significant difference between these two groups (P<0.05). These results suggest that compared with laminectomy fixation, the incidence of C5 nerve root palsy was lower after the laminoplasty with mini-trianium plate fixation, which can be widely used in decompression repair treatment of multilevel cervical spondylotic myelopathy.