Objective:To observe any clinical effect of supplementing ultrasound stimulation with balloon-guided injection of botulinum toxin into the upper esophageal sphincter in the treatment of cricopharyngeal achalasia.Methods:Forty patients with cricopharyngeal achalasia were randomly divided into an observation group and a control group, each of 20. Both groups were given routine swallowing rehabilitation training, while the observation group additionally had botulinum toxin injected into the upper esophageal sphincter guided by ultrasound and with the aid of balloon dilation. Before the experiment and after 2 weeks, both groups were evaluated videofluoroscopically and flexible endoscopic evaluation of swallowing was performed. Moreover, 2 weeks before the treatment and 2, 4 and 24 weeks afterward, everyone′s eating, leakage and aspiration, and oral and pharyngeal secretions were assessed using the functional oral intake scale (FOIS), the penetration-aspiration scale (PAS), the fiberoptic endoscopic dysphagia severity scale (FEDSS) and the Murray secretion scale (MSS).Results:After 2 weeks the average PAS, FEDSS and MSS scores of both groups had improved significantly, but the observation group′s averages[3(2, 5), 3(2, 5) and 2(1, 2)] were significantly better than those of the control group. 2, 4 and 24 weeks after the experiment the average FOIS scores of both groups also showed significant improvement, with the observation group′s average[3(2, 4), 4(2, 6) and 6(3, 7)] again significantly better than that of the control group.Conclusions:A botulinum toxin injection into the upper esophageal sphincter can effectively improve the swallowing of persons with cricopharyngeal achalasia with adequate safety and significant long-term benefits. Therefore, such treatment is worthy of clinical promotion and application.

Objective:To investigate the inhibitory effect of ferulic acid on the retina of diabetic mice and high glucose-induced human retinal pigment epithelium (RPE) cell injury and the mechanism.Methods:Thirty 8-week-old SPF male type 2 diabetic db/db mice were selected and divided into a model group and a ferulic acid group by the random number table method, with 15 mice in each group.Another 15 db/m mice of the same age were selected as a control group.The model and control groups received normal saline (5 ml/kg) by gavage daily, and the ferulic acid group received ferulic acid solution (0.05 g/kg) by gavage daily.After two months of treatment, the mice were sacrificed and the eyeballs were removed.The morphological changes of mouse retinal tissues were observed by hematoxylin-eosin staining.The fluorescence intensity and expression levels of mitochondrial calcium uniporter (MCU), p38 mitogen-activated protein kinase (p38 MAPK) and phosphorylated p38 MAPK (p-p38 MAPK) in mouse retinal tissues were detected by immunofluorescence staining and Western blot.Human RPE cells were divided into control group, dimethyl sulfoxide (DMSO) group, high glucose group and high glucose+ ferulic acid group.The control group received no treatment, and the other cell groups were cultured with the corresponding reagents for 24 hours.The reactive oxygen (ROS) level of RPE cells in each group was detected with the ROS detection kit.The mitochondrial membrane potential level of RPE cells was detected with the a mitochondrial membrane potential detection kit (JC-1).The MCU and microfilament fluorescence intensity of RPE cells were detected with the a microfilament green fluorescent probe.To explore the regulatory relationship between MCU, p38 MAPK and p-p38 MAPK, the MCU protein level was silenced and overexpressed by lentivirus transfection technology.The fluorescence intensity and expression levels of MCU, p38 MAPK and p-p38 MAPK proteins in RPE cells were detected by immunofluorescence staining and Western blot.The use and feeding of experimental animals followed the 3R principle and the Statement of the Association for Research in Vision and Ophthalmology on the Use of Animals in Ophthalmology and Vision Research.This study protocol was approved by the Ethics Committee of Ningxia Eye Hospital, People's Hospital of Ningxia Hui Autonomous Region (No.2019085).Results:The intercellular space of the outer nuclear layer, inner nuclear layer and ganglion cell layer of the retinal tissue in the model group was increased and the cell arrangement was disordered compared with the control group, and the retinal tissue in the ferulic acid group was significantly improved.Compared with the control group, the fluorescence intensity of MCU, p-p38 MAPK and MCU+ p-p38 MAPK protein of mouse retinal tissue in model group and ferulic acid group was significantly increased (all at P<0.05).Compared with the model group, the fluorescence intensity of MCU, p-p38 MAPK and MCU+ p-p38 MAPK protein of mice retinal tissue in ferulic acid group was significantly decreased (all at P<0.05).Compared with the control group, the relative expression levels of MCU, p38 MAPK and p-p38 MAPK proteins of mouse retinal tissue in model group were significantly increased (all at P<0.05).Compared with the model group, the relative expression levels of MCU, p38 MAPK and p-p38 MAPK proteins of mice retinal tissue in ferulic acid group were significantly decreased (all at P<0.05).The ROS fluorescence intensities in the control group, DMSO group, high glucose group and high glucose+ ferulic acid group were 0.22±0.02, 0.22±0.03, 0.30±0.02 and 0.24±0.02, respectively, and the overall difference was statistically significant ( F=7.845, P<0.01).The ROS fluorescence intensity was significantly higher in the high glucose group than in the control and DMSO groups, and it was significantly lower in the high glucose+ ferulic acid group than in the high glucose group (all at P<0.05).The mitochondrial membrane potential was significantly lower in high glucose group and high glucose+ ferulic acid group than in control and DMSO groups, and significantly higher in high glucose+ ferulic acid group than in high glucose group (all at P<0.05).Compared with the control group and DMSO group, the fluorescence intensity of MCU was higher in the high glucose group, accompanied by the decrease and thinning of cell microfilaments, and the fluorescence intensity of MCU protein was significantly decreased in high glucose+ ferulic acid group, with the number of microfilaments increased significantly.Compared with the control group and DMSO group, the fluorescence intensity and relative expressions of MCU, p38 MAPK and p-p38 MAPK proteins were significantly increased in the high glucose group (all at P<0.05).Compared with the high glucose group, the fluorescence intensity and relative expressions of MCU, p38 MAPK and p-p38 MAPK proteins were significantly decreased in the high glucose+ ferulic acid group (all at P<0.05).Compared with the control group and the empty vector group, the relative expressions of MCU, p38 MAPK and p-p38 MAPK proteins were significantly increased in the MCU overexpression group and significantly decreased in the MCU shRNA group and the MCU overexpression+ ferulic acid group (all at P<0.05).Compared with MCU overexpression group, the relative expressions of MCU, p38 MAPK and p-p38 MAPK proteins were significantly decreased in MCU shRNA group and MCU overexpression+ ferulic acid group, and the differences were statistically significant (all at P<0.05). Conclusions:Ferulic acid can regulate oxidative stress and mitochondrial dysfunction, thereby ameliorating retinal damage and high glucose-induced RPE cell injury in diabetic mice, which may play a protective role through MCU and p38MAPK signaling pathways.

ObjectiveTo screen out the mRNAs involved in the resistance of hepatoma cells to anlotinib using ceRNA microarray. MethodsHigh-dose shock combined with low-dose induction was used to culture hepatoma cells resistant to anlotinib, and CCK8 assay was used to verify the difference in the proliferation of drug-resistant hepatoma cells treated by anlotinib. The ceRNA microarray was used to screen out the differentially expressed genes between drug-resistant hepatoma cells and normal hepatoma cells, and real-time PCR was used to verify the differentially expressed genes detected by some microarrays. the independent samples t-test was used for comparison of continuous data between two groups, and the Kaplan-Meier method was used to analyze the overall survival of hepatoma cells samples, and the log-rank test was used to compare survival rates. Fisher’s exact test was used for chip screening. ResultsThere was a significant difference in gene expression between drug-resistant hepatoma cells and normal hepatoma cells, and 10 genes with the greatest difference were screened out for analysis by reducing the range. There were 4 genes associated with drug resistance and tumor growth, i.e., BIRC2, BIRC7, ABCC2, and MAPK8. There were significant reductions in the expression levels of BIRC2, ABCC2, and MAPK8 (P=0001 4, 0001 2, and 0.011 8), and there was a significant increase in the expression of BIRC7 (P＜0.001). The results of real-time PCR were consistent with those of microarray (t=10.74,32.65,18.34, and 2.80; P=0.000 4, 0.000 1, 0.000 1, and 0.044 8). The high expression of BIRC7 and the low expression of MAPK8 were associated with the significant reduction in survival time (P=0.022 0 and 0.005 6). ConclusionBIRC2, BIRC7, ABCC2, and MAPK8 are differentially expressed between anlotinib-resistant hepatoma cells and normal hepatoma cells and may be involved in the resistance of hepatoma cells to anlotinib.

Objective:To explore the related factors for clinical prognoses of ruptured anterior communicating artery (ACoA) aneurysms.Methods:A retrospective study was performed on the clinical data of 309 patients with ruptured ACoA aneurysms admitted to our hospital from January 2014 to January 2020. The preoperative data included age, gender, smoking history, hypertension, Hunt-Hess grading, Fisher grading, sizes of aneurysms, and spasm of parent artery; and the postoperative data included pneumonia, intracranial infection, cerebral hernia, recurrence and re-hemorrhage of aneurysms, and delayed cerebral ischemia. Clinical prognoses were assessed by modified Rankin scale (mRS). Univariate analysis and multivariate Logistic regression analysis were used to determine the independent risk factors for clinical prognoses. Preoperative model (independent risk factors appeared before surgery) and postoperative model (independent risk factors appeared during the whole treatment process) were constructed; based on these Logistic models, the preoperative and postoperative independent risk factors were concluded. Independent risk factors presented in the preoperative and postoperative models were used as variables to analyze the predictive value of the models by receiver operating characteristic (ROC) curve.Results:Among 309 patients, 264 (85.4%) had good prognosis and 45 (14.6%) had poor prognosis. (1) Univariate analysis showed that significant differences were noted in proportion of smoking patients, and patients with hypertension, Hunt-Hess grading IV-V, Fisher grading IV, wide-necked aneurysm, re-hemorrhage of aneurysms, cerebral vasospasm, pneumonia, intracranial infection, cerebral hernia, delayed cerebral ischemia, and postoperative lumbar cistern drainage between good prognosis group and poor prognosis group ( P<0.05). (2) Multivariate Logistic regression analysis showed that Hunt-Hess grading Ⅳ-V ( OR=24.198, P=0.000, 95%CI: 4.288-136.559), Fisher grading Ⅳ ( OR=4.792, P=0.044, 95%CI: 1.040-22.079), spasm of parent artery ( OR=12.136, P=0.005, 95%CI: 2.121-69.426), pneumonia ( OR=8.177, P=0.018, 95%CI: 1.438-46.506), postoperative cerebral hernia ( OR=147.042, P=0.002, 95%CI: 6.386-3385.519) and delayed cerebral ischemia ( OR=606.720, P=0.000, 95%CI: 52.288-7040.088) were independent risk factors for prognoses; postoperative lumbar cister drainage ( OR=0.072, P=0.050, 95%CI: 0.005-1.000) was the independent protective factor. (3) ROC curve showed that the preoperative model (with Hunt-Hess grading IV-V, Fisher grading Ⅳ and cerebral vasospasm as variables) had excellent discrimination with an area under the curve (AUC) of 0.870 ( 95%CI: 0.82-0.93, P=0.000), and the postoperative model (with variables of preoperative model, pneumonia, delayed cerebral ischemia, and herniation as variables) had excellent discrimination (AUC=0.980, 95%CI: 0.97-0.99, P=0.000). Conclusion:Besides decreasing Hunt-Hess grading and Fisher grading, and relieving the arterial spasm, the management of lumbar subarachnoid continuous drainage and avoidance of postoperative complications, such as cerebral hernia, delayed cerebral ischemia and pneumonia, can also play important roles in improving the prognoses of ruptured ACoA aneurysms.

[Abstract] Objective: To investigate the effect of metformin on the senescence-associated secretory phenotype (SASP) of doxorubicin-induced gastric cancer BGC823 cells. Methods: Human gastric cancer BGC823 cells were cultured in vitro and treated with doxorubicin at gradient concentrations (50, 100, 150 and 200 nmol/L). Cell senescence was detected by SA-β-gal staining, and SASP factor expression was detected by ELISA. The effects of metformin on cell senescence and SASP factor secretion induced by doxorubicin (100 nmol/L) were observed by adding gradient concentrations of metformin (0, 5, 10 and 20 mmol/L). Results: With the increase of doxorubicin concentration and treatment time, the senescence rate of gastric cancer BGC823 cells increased first and then decreased. At 96 h after 100 nmol/L doxorubicin treatment, the peak aging rate reached 68.7%, accompanied with significantly increased expressions of SASP factors IL-1a, IL-6, IL-8 and CXCL1. The proportion of senescent cells was (55.2±1.9)%, (48.7±2.2)% and (40.8±2.3)% respectively under the effects of 5, 10 and 20 mmol/L metformin, which was significantly lower than that in the non-metformin treatment group (P< 0.01). At the same time, with the increase of metformin concentration, the production of SASP factors IL-1α, IL-6, IL-8 and CXCL1 showed a gradient decline. Compared with the non-metformin treatment group, IL-6 and IL-8 decreased significantly under the effect of metformin above 10 mmol/L (P<0.05 or P<0.01), while IL-1α and CXCL1 decreased significantly under the effect of 20 mmol/L metformin (all P<0.05). Conclusion: Metformin can inhibit the senescence and SASP production of gastric cancer cells induced by doxorubicin.

Objective? To investigate the mastery of antiplatelet drugs knowledge among coronary heart disease patients with percutaneous coronary intervention (PCI), and to analyze its influencing factors. Methods? From November 2017 to February 2018, we selected 75 PCI patients with routine reexamine of department of cardiology at a classⅢgrade A hospital in Shanghai as subjects by convenience sampling. All of the patients were investigated with the self-designed antiplatelet drugs knowledge questionnaire for coronary heart disease patients with PCI. Results? Among 75 PCI patients, the total score of antiplatelet drugs knowledge among PCI patients was (30.64±6.90) with 54.71% for the scoring rate. Single factor analysis showed that there were statistical differences in the scores of antiplatelet drugs knowledge among PCI patients with different operative time, marital status and monthly incomes (P＜ 0.05). Conclusions? Mastery of antiplatelet drugs knowledge questionnaire among coronary heart disease patients with PCI needs to be improved. Pay attention to the influences of operative time after PCI, marital status and monthly incomes on mastery of antiplatelet drugs knowledge. We should improve the mastery of antiplatelet drugs knowledge by constructing health education evaluation system, strengthening health education and perfecting follow-up system to guide patients to take medicine correctly and regularly, so as to reduce the death risk.

In order to establish the minipig model of chronic myocardial ischemia and apply non-invasive telemetry technique, the minipig model of chronic myocardial ischemia was made induced by Vitamin D3, isoproterenol and combined with high fat diet, and the non-invasive telemetry technique was used to detect and evaluate the symptoms of minipig model of chronic myocardial ischemia.Moreover, the effects of transport stress and the risk factors of atherosclerosis (AS) induced by high fat diet among Wuzhishan minipigs, Bama minipigs and Tibetan minipigs were also evaluated.Our study has successful established the Bama minipig model of chronic myocardial ischemia and the technical specification for evaluation,.The non-invasive telemetry technique can be used to detect and evaluate the symptoms of chronic myocardial ischemia model, and defines minipigs at least need to keep for more than 4 weeks after transport stress to adaptive recovery period.In addition, the different characteristics of AS risk factors such as hyperlipidemia, hypertension and hyperinsulinemia were observed in Wuzhishan minipigs, Bama minipigs and Tibetan minipigs in high fat environment, and this provides a reference for the selection and application of minipigs in the research of cardiovascular diseases.

Adenoma, Liver Cell ; Gallbladder Diseases ; Histiocytosis, Sinus ; Humans ; Liver Neoplasms

Objective To investigate the relationship between apolipoprotein E ( ApoE ) gene polymorphism and cerebral infarction patients with different gender and etiological typing. Methods A total of 91 patients with cerebral infarction aged≥60 years ( cerebral infarction group) were enrolled. They were divided into either a large artery atherosclerotic (LAA) stroke group (n=37) or a small artery occlusion (SAO) stroke group (n=54) according to the Trial of Org 10172 in acute stroke treatment (TOAST) classification. A total of 105 age-,sex-,and residence-matched healthy subjects were enrolled as controls. A Nested Allele-Specific Multiplex Polymerase Chain Reaction Method was used to detect the ApoE gene polymorphism. The ApoE gene polymorphism of cerebral infarction of different gender and etiological typing were compared. Results ( 1 ) ApoE Genotypes of E2/2, E2/3, E2/4, E3/3, and E3/4 were detected,but the ApoE E4/4 was not detected. (2) There were no significant differences in the frequencies of ApoE genotypes and each gene carrier frequency between the cerebral infarction group and the control group (all P>0. 05). There was significant difference in ApoE genotype frequencies and each gene carrier frequency of the males between the cerebral infarction group and the control group (P<0. 01,P<0. 05). Both the E3/3 genotype frequency (56. 1%) and ε3 carrier frequency (78. 0%) of the cerebral infarction group were lower than the males of the control group ( 79. 2% and 89. 6% respectively );both the E3/4 genotype frequency (31. 7%) and ε4 carrier frequency (15. 9%) were higher than the control group (7. 5% and 3. 8%respectively). There was no significant differences in the ApoE genotype frequency and gene carrier frequency in female participants between the two groups (all P>0. 05). (3) There were no significant differences in the ApoE genotype frequency and gene carrier frequency among the LAA,SAO,and control groups. There was significant difference in the ApoE genotype frequency and gene carrier frequency in males between the LAA group and the control group (P>0. 01);the genotype frequencies of E2/3 and E3/E3 (6. 7% and 46. 7%),ε2,as well as theε3 carrier frequency (3. 3% and 73. 3%) of LAA were lower than those of the control group (13. 2%,79. 2%,6. 6%,and 89. 6%,respectively);the E3/4 genotype frequency andε4 carrier frequency of the LAA subtype were 46. 7% and 23. 3% respectively. They were all higher than 7. 5% and 3. 8% in the control group. However,there were no significant differences in males among the SAO group,the control group,and the 3 groups of females ( the LAA subtype,SAO subtypes,and the control group) (P>0. 05). Conclusion ε4 gene may be a risk factor for LAA in males. The association of ApoE gene polymorphism with cerebral infarction in females is not found.

Objective To analyze the genetic variation among white hair black eyes (WHBE) rabbit, Japanese white ( JW) rabbit and New Zealand white ( NZW) rabbit using random amplified polymorphic DNA ( RAPD) technique . Methods Thirty rabbits (male/female 1∶1) of each strain were used in this study.The genomic DNA was extracted from 90 rabbits.Sixty arbitrary primers were used to amplify DNA of rabbits with RAPD-PCR method.Based on the preliminary experiments , polymorphic primers were selected to analyze the genetic variation among the three rabbit strains .The experi-mental data were analyzed using Popgene 3.2 software.Results (1) Twenty-five polymorphic primers were selected among 60 arbitrary primers.493 amplified fragments were detected ranging from 100 bp to 1800 bp.Sixteen primers among 25 arbitrary primers could not only amplify the common DNA bands of 3 rabbit breeds , but also amplify particular alleles in the WHBE rabbit.(2) 234 RAPD sites were detected by agarose gel electrophoresis in WHBE rabbit , among which 166 sites were polymorphic , accounting for 70.94%.228 RAPD sites were detected by agarose gel electrophoresis in the JW rabbit, while 122 sites of them were polymorphic , accounting for 53.51%.231 RAPD sites were detected by agarose gel e-lectrophoresis in the NZW rabbits , with 94 sites being polymorphic, accounting for 40.69%.(3) The Shannon genetic di-versity index of WHBE rabbit, JW rabbit and NZW rabbit was 0.3385, 0.2222 and 0.1905, respectively.(4) The genet-ic similarity between JW rabbit and NZW rabbit was highest among the three rabbit breeds (0.8443), followed by that be-tween WHBE rabbit and JW rabbit (0.8204), and the genetic similarity between WHBE rabbit and NZW rabbit (0.7862) was the lowest .Conclusions Our results demonstrate that there are both genetic similarities and genetic variations among WHBE rabbit, JW rabbit and NZW rabbit .The RAPD technique can be used to delect the genetic relationships among dif-ferent breeds and different individuals of the same breed of rabbits .