Objective To investigate the influences of grape seed proanthocyanidin extract (GSPE) on cardiovascular system, nervous system and respiratory system of experimental animals, and provide general pharmacological data for further research and application. Methods The influences of GSPE on blood pressure, heart rate, electrocardiogram, breathing frequency and tidal volume in anesthetic dogs after duodenal administration were observed, the impacts on spontaneous activity, coordinated motion, and the sleep situation with threshold dose and subthreshold dose of pentobarbital sodium in mice after intragastric administration were observed. Results GSPE showed no side effects on blood pressure, heart rate, electrocardiogram, breathing frequency and tidal volume in anesthetic dogs at the dosage of 857.00, 214.29, 42.86 mg/kg (P>0.05). At the dosage of 428.57, 214.29, 42.86 mg/kg, GSPE had no obvious influence on spontaneous activities and coordinated movements in mice (P>0.05). GSPE did not evidently change the number of sleeping animals, the sleep latency and the sleeping duration with subthreshold dose and threshold dose of pentobarbital sodium (P>0.05). Conclusion GSPE has no evident adverse effects on central nervous system, cardiovascular system and respiratory system in animals.

BACKGROUND: It has been reported that in China, human bone marrow mesenchymal stem ceils are mostly harvested from adults. Studies on bone marrow mesenchymal stem cells in children are few.OBJECTIVE: To isolate and expand bone marrow mesenchymal stem cells from children, and to analyze the biological characteristics of bone marrow mesenchymal stem cells and their potential of differentiating into osteoblasts, adipocytes and neural like cells.DESIGN: Observational comparative study.SETTING: Tongji Medical College, Huazhong University of Science and Technology.MATERIALS: Experiments were performed at the Laboratory of Department of Orthopaedics of Wuhan Tongji Hospital from March to September 2006. Bone marrow mesenchymal stem cells were collected from one boy patient and two girl patients aged 5-8 years, who received pelvis osteotomy for dysplasia of the hip joint. The experimental procedures were approved by the Hospital Ethics Committee and family members of all children patients singed the informed consent.Dexamethasone, vitamin C, β-sodium glycerophosphate, 3-1sobutyl-1-methylxanthine, insulin, indometacin and butylated hydroxyanisole were bought from Sigma Company. Dimethyl sulphoxide was purchased from Amersco Company.METHODS: Bone marrow mesenchymal stem cells were cultured from mononuclear cells isolated over a Percoll gradient.Bone marrow mesenchymal stem cells were observed under an inverted phase contrast microscope. Bone marrow mesenchymal stem cells could differentiate into osteoblasts, adipocytes and neural like cells with osteoblast inductor (β-sodium glycerophosphate, dexamethasone, vitamin C), lipoblast inductor (dexamethasone, 3-isobutyl-1-methylxanthine,bovine insulin, indometacin) and serum-free medium inductor (dimethyl sulphoxide, butylated hydroxyanisole) respectively.Osteoblast marker (alkaline phosphatase, osteocalcin mRNA, calcium node), adipocyte marker (lipid droplet, PPAR γ-2mRNA) and neural ceil-like marker (nissl body, neuron specific enolase, neurofilament protein) were respectively determined by the immunohistochemical method, polymerase chain reaction and immunocytochemical method.MAIN OUTCOME MEASURES: ①Appearance and proliferation of bone marrow mesenchymal stem ceils from children,and ②determination results of osteoblast, adipocyte and neural cell markers.RESULTS: ①Children bone marrow mesenchymal stem cells could easily adhere to the wall, appeared fusiform, had high reproductive activity and arranged vortically after fusing. ②Appearance of bone marrow mesenchymal stem cells changed after receiving inductor. Osteoblast marker, adipocyte marker and neural cell-like marker were positive after chemical staining, polumerase chain reaction and immunocyte staining.CONCLUSION: Children bone marrow mesenchymai stem cells show stable proliferation, passage and multi-direction differentiation towards osteoblasts, adipocytes and neural like cells.

Objective To observe the effects of Marigold lutein on the cardiovascular system, the nervous system and the respiratory system of experimental animals. Methods The influence on blood pressure, heart rate, electrocardiogram, breathing flow and frequency in anesthetic dogs were observed after duodenum euonymus, and the impact on spontaneous activity, coordinated movement, and the sleep situation of threshold dose and sunthreshold dose pentobarbital sodium in mice were observed by intragastric administration of Marigold lutein. Results Marigold lutein had no side effects on breathing flow and frequency, blood pressure, heart rate, electrocardiogram in anesthetic dog, and had no obvious influence on spontaneous activity and coordinated movement in mice. Marigold lutein did not evidently change the number and the time of sleeping in mice. Conclusion Marigold lutein has no evident effects on respiratory system, cardiovascular system and central nervous system of experimental animals.

Objective To study the general pharmacological effects of Aloe's whole-leaf freeze-dried powder (AWFD), and observe its influence on cardiovascular system, nervous system and respiratory system of laboratory animals, so as to offer an experimental basis for clinical application. Methods Forty-eight mice were randomized into blank control group, high dosage group, medium dosage group and low dosage group of AWFD (12 mice for each group). AWFD high, medium and low dosage groups were treated by intragastric at the dose of 12.20, 3.90, 0.65 g/(kg?d), blank control group was treated by equal volume of sterilized distilled water. After three days, general behavior, spontaneous activity, coordinated movement, sleep situation induced by sodium pentobarbital in subthreshold dose and suprathreshold dose were observed. Twenty-four beagle were randomized into blank control group, high dosage group, medium dosage group and low dosage group of AWFD (6 beagles for each group). AWFD high, medium and low dosage groups were treated by duodenum at the dose of 6.10, 3.41, 0.71 g/(kg?d), blank control group was treated by equal volume of sterilized distilled water. The influence on blood pressure, heart rate, electrocardiogram, breathing flow and frequency in anesthetic dogs were observed. Results Three dosages of AWFD had no obvious influence on spontaneous activity and coordinated movement in mice, and had no evidently influence on sleep number and duration, but the high dosage group of AWFD had influence on sleep latency (P<0.01). AWFD had no impact on blood pressure, heart rate, electrocardiogram, breathing flow and frequency in anesthetic dogs. Conclusion AWFD has no evident effects on cardiovascular system and respiratory system in laboratory animal, however, the impact on the central nervous system remains to be further verified.

Objective: To investigate the expressionof proline-rich protein 11 (PRR11) in esophageal carcinoma (EC) tissues and to study it’s effect on the proliferation and metastasis of human EC TE-2 cells in vitro. Methods: Eighty patients were pathologically diagnosed with EC the Department of Thoracic Surgery of the Second Affiliated Hospital of Zhengzhou University from October 2016 to October 2018, and their surgically resected cancer tissues and corresponding para-cancerous tissues were collected for this study. qPCR was used to detect the expression of PRR11 mRNAin tissues or cells. Log-rank Test was used to analyzethe relationship between the expression of PRR11 in EC tissues and general data, histological type, lymphatic metastasis, depth of invasion and TNM stageof the EC patients. Kaplan-Meierplot was used to analyze the association between PRR11 mRNA and patients’prognosis. TE-2 cells were transfected with lentivirus shRNA to construct cell line with PRR11 knockout and corresponding control cell lines, as shPRR11#1, shPRR11#2 and control group. qPCR and WB assays were used to verify the mRNA and protein expressions of PRR11 in cell lines respectively. MTT was used to examine the proliferation of transfected cells, and Transwell experiments were used to detect cell invasion and migration. Results: The expression of PRR11 mRNA in EC was higher than that in para-cancer tissues (P<0.05). There was significant correlation between PRR11 over-expression and histological type, lymphatic metastasis, depth of invasion and TNM stage(all P <0.05), and high PRR11 expression was significantly related with the poor prognosis of EC patients (P<0.05). The mRNA and protein expressions of PRR11 in cells of shPRR11#1 and shPRR11#2 groups were significantly lower than those in control group (all P <0.05). MTT assay showed that the proliferation of cells in shPRR11#1 and shPRR11#2 groups was significantly lower than that in the control group (P<0.05 or P<0.01). The results of Transwell invasion and migration assays showed that the average number of cells with in each field of viewin shPRR11#1 and shPRR11#2 groups was significantly lower than that in the control group (P<0.01). Conclusion: PRR11 is over-expressed in EC tissues and PRR11 over-expression is closely related to the occurrence, progression and prognosis of esophageal cancer. In vitro experiments have also demonstrated that knockdown of PRR11 can inhibit the proliferation, invasion and migration of EC. PRR11 can be used as a potential molecule marker and drug targets for EC. ··

Objective:To investigate the effectiveness and advantage of visual laryngoscope in the treatment of patients with sudden cardiac arrest who need spontaneous respiration tracheal intubation.Methods:Totally 60 patients who suffered from cardiac arrest and needed spontaneous respiration tracheal intubation were enrolled from June 2020 to February 2021 in the Affilicated Hospital of Chifeng University. Theywere randomlydivided into two groups-traditional laryngoscope (TL) group and visual laryngoscope (VL) group, with30 patients in each group. Then the success rate of glottis exposure, the operative time, success rate and complication rate of tracheal intubation were compared between the two groups. Subgroup assessment between the junior emergency resident doctor (A group) and the senior emergency resident doctor (B group) was conducted.Results:According to Cormack Lehan grades, the success rate of glottis exposure in VL group was higher than that in TL group ( P>0.05), and the success rate of Grade I in VL group was significant higer than that in TL group: 56.70%(17/30) vs. 30.00%(9/30), P<0.05. The trial times of successful intubation cases and the operative time of successful intubation cases were significantly less than those in TL group (1.30 vs 1.67, P = 0.049) and (56.37 s vs 67.12 s, P<0.05). In the subgroup, the one-time success rate of tracheal intubation in A-TL group was significantly lower than that in B-TL group (4/15 vs. 11/15, P<0.05), while the one-time success rate of tracheal intubation in A-VL group was 60.00%, which is lower than that in B-VL group ( P>0.05). The operative time consumed for successful intubation in A-TL group was significantly longer than that in B-TL group: 78.00 s vs. 55.57 s, P<0.05, while the operative time in A-VL group was a little longer than that in B-VL group ( P>0.05). Conclusions:Visual laryngoscope used in spontaneous respiration tracheal intubation can not only increase the success rate of glottis exposure, decrease trial times and shorten operative time of intubation, but also improve the success rate and decrease the complication rate of emergency tracheal intubation.

Objective:To develop a novel, flexible, dual-arm, master-slave digestive endoscopic minimally invasive surgical robot system named dual-arm robotic endoscopic assistant for minimally invasive surgery (DREAMS) and to evaluate its feasibility for endoscopic submucosal dissection (ESD) by using ex vivo porcine stomachs.Methods:A novel endoscopic robot (DREAMS) system was developed which was composed of a flexible two-channel endoscope, two flexible robotic manipulators, a master controller, a robotic arm, and a control system. A total of 10 artificial round-like lesions with diameters ranging from 15 to 25 mm were created (5 in gastric antrum and 5 in gastric body) by using fresh peeled stomach of healthy pigs as the model. Submucosal dissection was performed with the assistance of the DREAMS system by two operators. The main outcome was submucosal dissection speed, and the secondary outcomes included muscular injury rate, perforation rate, and grasping efficiency of the robot.Results:All 10 lesions were successfully dissected en bloc by using the DREAMS system. The diameter of the artificial lesions was 22.34±2.39 mm, dissection time was 15.00±8.90 min, submucosal dissection speed was 141.79±79.12 mm 2/min, and the number of tractions required by each ESD was 4.2 times. Muscular injury occurred in 4/10 cases of ESD. No perforation occurred. Conclusion:The initial animal experiment shows the DREAMS system is safe and effective.

Myocardial ischemia-reperfusion injury （MIRI） is a serious complication of revascularization in patients with myocardial infarction. The nuclear factor erythroid 2-related factor 2 （Nrf2）/heme oxygenase-1 （HO-1） signaling pathway plays an important role in the pathological process of MIRI. Currently，research has found that traditional Chinese medicine has a good effect on myocardial injury caused by ischemia-reperfusion. Based on the Nrf2/HO-1 signaling pathway，this article summarizes the action mechanism of traditional Chinese medicine formulas and monomers in intervening with MIRI. It is found that traditional Chinese medicine formulas （Yixin formula，Wenyang tongmai formula，Dingxin formula Ⅰ），monomers such as terpenoids （ginkgolides， astragaloside Ⅳ，ginsenosides），phenols （brazilin，hematoxylin A，resveratrol） and quinones （aloe，emodin） can alleviate MIRI by activating the Nrf2/HO-1 signaling pathway，inhibiting oxidative stress and inflammatory reactions，etc.

ObjectiveTo investigate the mechanism of anti-pulmonary fibrosis of cannabidiol by ultra-high performance liquid chromatography-quadrupole-time-of-flight mass spectrometry（UPLC-Q-TOF-MS）. MethodSD rats were randomly divided into blank group， model group， prednisone group（3.15 mg·kg^-1） and cannabidiol low， medium and high dose groups（12， 36， 108 mg·kg^-1）， with 8 rats in each group. The rat model of pulmonary fibrosis was established by intratracheal injection of bleomycin（5 mg·kg^-1）， which was administered continuously for 28 days after successful modeling. The pathological changes of rat lung tissue were observed， and enzyme-linked immunosorbent assay（ELISA） was used to detect the expression levels of matrix metalloproteinase-7（MMP-7）， type Ⅱ alveolar cell surface antigen（KL-6）， pulmonary surfactant-associated protein A（SP-A） and SP-D in serum. The expression levels of type Ⅰ collagen（Col-Ⅰ） and fibronectin（FN） in lung tissues were detected by immunohistochemistry， and the expression of mucin 5 subtype AC（MUC5AC） was detected by immunofluorescence. UPLC-Q-TOF-MS was used to search for potential biomarkers and related metabolic pathways of cannabidiol in treating pulmonary fibrosis. ResultCompared with the blank group， there were a large number of inflammatory cell infiltration and continuous fibrosis lesions in the lung tissue of rats in the model group. Compared with the model group， the inflammatory infiltration and blue collagen deposition in the lung tissue of rats in the prednisone and cannabidiol groups were reduced. Compared with the blank group， the expressions of MMP-7， KL-6， SP-A and SP-D in serum of the model group were significantly increased（P<0.01）， while the expressions of MMP-7， KL-6， SP-A and SP-D in the prednisone and cannabidiol high dose groups were significantly decreased by comparing with the model group（P<0.05， P<0.01）. Compared with the blank group， the expression levels of Col-Ⅰ and FN in the lung tissues of the model group were significantly increased， and the fluorescence intensity of MUC5AC was significantly increased（P<0.01）. Compared with the model group， the expression levels of Col-Ⅰ and FN in the lung tissues of the prednisone and cannabidiol high dose groups were significantly decreased（P<0.05， P<0.01）， and the expression of MUC5AC was significantly decreased（P<0.01）. Compared with the blank group， a total of 18 differential compounds were screened out in the model group， which could be used as potential biomarkers， and cannabidiol could call back 16 of them， mainly involving 4 metabolic pathways（linoleic acid metabolism， phenylalanine， tyrosine and tryptophan biosynthesis， arachidonic acid metabolism， and niacin and niacinamide metabolism）. Compared with the blank group， the relative contents of potential biomarkers arachidonic acid and linoleic acid were significantly increased in the model group（P<0.05， P<0.01）， while the relative contents of 5，6-EET， L-tyrosine and niacinamide were significantly decreased（P<0.01）. Compared with the model group， cannabidiol could significantly reduce the relative contents of arachidonic acid and linoleic acid， and significantly increase the relative contents of 5，6-EET， L-tyrosine and niacinamide（P<0.01）. ConclusionCannabidiol has an intervention and remission effect on pulmonary fibrosis， and its mechanism may be related to linoleic acid metabolism， phenylalanine， tyrosine and tryptophan biosynthesis， arachidonic acid metabolism， niacin and niacinamide metabolism.