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Objective To investigate the role of pcgf5a gene during zebrafish embryonic development. Methods pcgf5a morpholine antisense morpholino oligomers(MO) was microinjected to delete the expression of pcgf5a gene, and possible phenotypes were examined in pcgf5a-deficient embryos. The whole mount in situ hybridization and Real-time PCR were used to detect alterations of key genes related to the development of nervous system and eye. Results After pcgf5a gene knockdown, the development of ectoderm and mesoderm of zebrafish embryos were affected. The brain size became smaller, and the eyes developed retarded, and the tail was curled and the body axis was shortened(548/891). In addition, the expressions of sox2(82/98), sox3(73/84), foxg1(70/88), pax6a(36/45), pax2a, vsx2 and rx1(n = 5) were significantly reduced. Conclusion pcgf5a affects the morphogenesis of nervous system and eye, possibly due to regulating the expression of the transcription factors related to their development via the processes of cell proliferation and apoptosis.
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[Abstract] Objective To investigate the effect of 6-gingerol treatment on cognitive behavior after hypoxic-ischemic brain injury (HIE) in neonatal mice, and to explore the protective mechanism of 6-gingerol on HIE brain injury in neonatal mice by observing the effects on neuronal survival and neural stem cell proliferation. Methods The right common carotid artery was ligated in Kunming mice (78) on the 7th day after birth and HIE model was established after 90 minutes of hypoxic treatment. 6-gingerol was injected intraperitoneally. The cognitive behavior was detected by Morris water maze test; 2,3,5-triphenyl tetrazolium chloride (TTC) staining was used to observe the changes of brain injury; The changes of synaptic structure and number were obseved by transmission electron microscopy; HE staining, Nissl staining and dihydroethidium(DHE) staining were used to observe the pathomorphological changes of hippocampus in each group; The proliferation of neural stem cells and the expression of related transcription factors were detected by immunofluorescence and Real-time PCR; The changes of Akt signal pathway were detected by Western blotting. Results 6-gingerol treatment could improve the long-term learning and memory ability, reduce the brain injury and brain edema of neonatal mice after HIE, and improve synaptic plasticity of mice after HIE. In the 6-gingerol treatment group, the disorder of hippocampal cells in the diseased side of HIE was improved, the number of necrotic cells decreased, the proliferation ability of hippocampal neural stem cells and the expression levels of nestin and sex determining region box transcription factor 2 (Sox2) related transcription factors increased significantly, and the level of phosphorylated Akt (p-Akt) increased. Conclusion It is found that 6-gingerol can improve the learning and memory ability of HIE mice in adulthood and reduce brain tissue injury after HIE. 6-gingerol may play a role in inhibiting the production of reactive oxygen species(ROS), reducing neuronal injury and upregulating the expression of Akt signal pathway, promoting the proliferation of hippocampal neural stem cells, so as to provide potential drugs for the treatment of neonatal HIE.
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Objective To explore the effect and mechanism of ginsenoside Rb1 on the repair of sciatic nerve injury (SNI) in mice. Methods Seventy-eight adult male Kunming mice were randomly divided into sham group (26), SNI group (26), SNI+Rb1 group (26). The SNI+Rb1 group was given 10 mg/kg ginsenoside Rb1 (i.p.), and the SNI group and the sham group were given the same volume of normal saline. The injury method was established by squeezing the sciatic nerve. Sciatic functional index (SFI) was used to evaluate sciatic nerve function. Growth associated protein 43 (GAP43) immunofluorescent staining was used to detect neural regeneration and repair on day 14, and the structure changes of the myelin sheath of the injured segment were observed under transmission electron microscope. Ki67 and S100β were used to detect the proliferation and migration ability of Schwann cells, and Real-time PCR was used to detect the mRNA expression levels after crush on day 3 and day 7. Results SFI of SNI+Rb1 group was higher than SNI group. The HE result showed that the sciatic nerve was uniform in the SNI + Rb1 group. The result of immunofluorescent staining displayed that Rb1 enhanced GAP43
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Objective: To observe the clinical efficacy of filiform fire-needling plus continuous passive motion (CPM) therapy for frozen shoulder. Methods: A total of 72 patients were randomized into an observation group and a control group by the random number table method, with 36 cases in each group. Patients in the control group received celecoxib capsule plus CPM, while those in the observation group received filiform fire-needling plus CPM. The whole course of treatment lasted for 2 weeks in both groups. The visual analog scale (VAS), Constant-Murley score (CMS) and range of motion of shoulder joint were measured for therapeutic efficacy evaluation. Results: The total effective rate was 91.7％ in the observation group, higher than 72.2％ in the control group, and the between-group comparison showed statistical significance (P<0.05). After treatment, the VAS scores in the two groups dropped significantly, the CMS as well as the range of motion including abduction, forward flexion and extension were all increased significantly, and the intra-group comparisons showed statistical significance (all P<0.05). The betweengroup comparisons showed the improvements in these items in the observation group were more significant than those in the control group (all P<0.05). Conclusion: Filiform fire-needling plus CPM can produce more significant efficacy than celecoxib capsule plus CPM for frozen shoulder; it can alleviate pain, improve shoulder function and restore joint range of motion in such patients.
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Objective: The effects of different decoction time and different decoction temperature on its chemical constituents of anthraquinones, anthrone, and tannins were revealed by pseudo-targeted metabolomics, which provided the basis for clinical use of rhubarb. Methods: Using LTQ-Orbitrap-MSn to accurately excavate the characteristic ions of various chemical components of rhubarb, the ion pairs of chemical components were determined as much as possible by characteristic ions to obtain the peak area of various components by using the MRM mode of QQQ-MS. The effects of different decoction methods on the chemical components of rhubarb was compared by multivariate statistical analysis combined with paired t test. Results: Both decoction time and decoction temperature have impacts on the chemical components of rhubarb decoction. Short-term decocting for 15 min was beneficial to the dissolution of dianthrone glycosides and anthraquinone glycosides, while long-term decocting for 60 min was beneficial to the dissolution of tannins; Compared with boiling water maceration, boiling water decoction was more favorable for the dissolution of anthraquinones and tannins. Conclusion: This paper adopts the method of pseudo-targeted metabolomics, and clearly points out that short-term decocting or maceration is conducive to exerting the laxative effect of rhubarb, and long-time boiling decocting is conducive to exerting clearing heat and detoxifying efficacy, which provides a reference for the clinical application of rhubarb.
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OBJECTIVE@#To analyze the genotypes and the hematological phenotypic characteristics of α-thalassemia in different areas of Fujian and to evaluate the values of mean corpuscular volume (MCV), mean corpuscular hemoglobin (MCH), hemoglobin (Hb), RBC distribution width/red blood cell (RDW/RBC) for screening α-thalassemia in this area.@*METHODS@#The Gap-PCR assay was applied for detecting 3 common deletional mutations of patients with α-thalassemia, and the reverse dot-blot (RDB) assay was adopted to detect the foci of 3 common non-deletional gene mutations.Then,the hematological parameters of individuals with α-thalassemia were analyzed. Finally, the optimal cut-off value in hematological indexes for screening α-thalassemia were determined by the ROC curve.@*RESULTS@#Altogether 16 types of gene mutations were found in 772 patients with α-thalassemia. Among them, the -SEA/αα deletion mutation was the most common which was observed in 521 cases(67.49%). Compared with the control group, the differences in MCV, MCH, and Hb were statistically significant between the patients of the same sex but no same type. In male groups, the RDW/RBC ratio was statistically significant in individuals of light type and HbH disease as compared with the healthy control group. But in female groups, the statistical different of RDW/RBC ratio was found between only HbH disease group and control group. MCV<81.25 fl, MCH<27.30 pg, Hb(male)<128.5 g/L, and Hb(female) <123.5 g/L, with the highest specificity and the highest sensitivity, were the best cut-off points for screening α-thalassemia in the laboratory.@*CONCLUSION@#Due to the difference of regional heterogeneity and hospital equipment environment, the different laboratories need to establish cut-off value for screening α-thalassemia suitable for its local region. In future, our laboratory can use MCV<81.25 fl, MCH<27.30 pg, Hb(male)<128.5 g/L, and Hb(female) <123.5 g/L for value for clinical screening, of α-thalassemia.
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Female , Humans , Male , China , Erythrocyte Indices , Genotype , Mass Screening , alpha-ThalassemiaABSTRACT
OBJECTIVE@#To observe the clinical efficacy of warm acupuncture combined with yoga posture method in the treatment of periarthritis with frozen period.@*METHODS@#Ninety patients with periarthritis who met the inclusion criteria were randomly divided into a control group 1, a control group 2 and an observation group, 30 cases in each group. Warm acupuncture was applied in the control group 1 (Jianzhen (SI 9), Jianyu (LI 15), Jianliao (TE 14), etc were selected), yoga posture method was applied in the control group 2, warm acupuncture combined with yoga posture method were given in the observation group, the treatment was given once a day, 10 times as a course with 2 days between courses and continuous for 2 courses. After 2 courses of treatment, the shoulder joint pain score and shoulder function grading were used to evaluate the clinical efficacy, and the clinical efficacy was observed.@*RESULTS@#①The pain scores of the three groups were significantly lower after treatment (all 0.05). ②After treatment, the functional classification of shoulder joints were significantly improved in the three groups (all 0.05). ③After 2 courses of treatment, the effective rate of the observation group was 86.7% (26/30), which was better than 70.0% (21/30) in the control group 1 and 76.7% (23/30) in the control group 2 (both <0.05).@*CONCLUSION@#Warm acupuncture combined with yoga posture method can effectively relieve shoulder pain and improve dysfunction. The clinical comprehensive effect is better than simple acupuncture and yoga posture method.
Subject(s)
Humans , Acupuncture Points , Acupuncture Therapy , Periarthritis , Therapeutics , Posture , Treatment Outcome , YogaABSTRACT
AIM:To investigate the relationship between adrenergic receptor(AR) R389G gene polymorphism and changes in intraocular pressure in patients with primary open angle glaucoma (POAG). METHODS: We selected 100 patients with newly diagnosed POAG from January 2015 to February 2017 in our hospital. The patient's intraocular pressure was measured at 6:00, 12:00, 18:00, and 24:00 daily and the mean value was calculated. Patients with mean intraocular pressure >30mmHg were included in Group A (44 patients), patients with mean intraocular pressure <30mmHg were treated as Group B (56 patients). The clinical data of two groups of patients were compared, and multivariate Logistic regression analysis was used to investigate the relationship between β1AR R389G gene polymorphism and changes in intraocular pressure in patients with POAG. RESULTS: The frequency of GG genotype in Group A (27.3%) was significantly higher than that in Group B (8 9%) (P < 0. 05). The allele G frequency was significantly higher in Group A (40.9%) than in Group B (18.8%), with a statistically significant difference (P<0 05). By multivariate Logistic regression analysis, increased systolic blood pressure and increased frequency of β1AR R389G allele were independent risk factors for increased intraocular pressure in POAG patients (P <0 05). CONCLUSION: The β1AR R389G gene polymorphism is closely related to POAG and is an independent risk factor for increased intraocular pressure in patients with POAG.
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Diagnostic ions filter method was used to rapidly detect and identify the phenolic compounds in Rheum palmatum based on ultra performance liquid chromatography coupled with quadrupole time-of-flight mass spectrometry (UPLC-Q-TOF/MSE). The representative authentic standards of phenolic compounds, including gallic acid, (+)-catechin, (－)-epicatechin, (－)-epicatechin-3-O-gallate and procyanidin B2, were subjected to analysis by UPLC-Q-TOF/MSE system with negative ion mode. Fragmentation patterns of each standard were summarized based on assigned fragment ions. The prominent product ions were selected as diagnostic ions. Subsequently, diagnostic ions filter was employed to rapidly recognize analogous skeletons. Combined with retention time, accurate mass, characteristic fragments and previous literature data, the structures of the filtered compounds were identified or tentatively characterized. A total 63 phenolic compounds (36 phenolic acid derivatives, 8 flavonoid derivatives and 19 tennis derivatives) in R. palmatum were identified, including 6 potential new compounds. The method of diagnostic ions filter could rapidly detect and identify phenolic compounds in R. palmatum This study provides a method for rapid detection of phenolic compounds in R. palmatum and is expected to complete the material basis of rhubarb.
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The improvement of antibiotic rational use in China was studied by usage analysis of combination antibiotic therapy for type I incisions in 244 hospitals. Five kinds of hospitals, including general hospital, maternity hospital, children's hospital, stomatological hospital and cancer hospital, from 30 provinces were surveyed. A systematic random sampling strategy was employed to select outpatient prescriptions and inpatient cases in 2011 and 2012. A total of 29 280 outpatient prescriptions and 73 200 inpatient cases from 244 hospitals in each year were analyzed. Data were collected with regards to the implementation of the national antibiotic stewardship program (NASP), the overall usage and the prophylactic use of antibiotic for type I incisions. Univariate analysis was used for microbiological diagnosis rate before antimicrobial therapy, prophylactic use of antibiotics for type I incision operation, and so on. For multivariate analysis, the use of antibiotics was dichotomized according to the guidelines, and entered as binary values into logistic regression analysis. The results were compared with the corresponding criteria given by the guidelines of this campaign. The antibiotic stewardship in China was effective in that more than 80% of each kind of hospitals achieved the criteria of recommended antibiotics varieties. Hospital type appeared to be a factor statistically associated with stewardship outcome. The prophylactic use of antibiotics on type I incision operations decreased by 16.22% (P<0.05). The usage of combination antibiotic therapy for type I incisions was also decreased. Region and bed size were the main determinants on surgical prophylaxis for type I incision. This national analysis of hospitals on antibiotic use and stewardship allows relevant comparisons for bench marking. More efforts addressing the root cause of antibiotics abuse would continue to improve the rational use of antibiotics in China.
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Female , Humans , Male , Anti-Bacterial Agents , Therapeutic Uses , China , Epidemiology , Drug Prescriptions , Drug Utilization , Hospitals , Inpatients , Outpatients , Surgical Wound , Epidemiology , Microbiology , TherapeuticsABSTRACT
<p><b>OBJECTIVE</b>To observe the expression of receptor activator of NF-κB ligand (RANKL) and its decoy receptor osteoprotegerin (OPG) during unloading period of dental implants.</p><p><b>METHODS</b>An animal model of dental implants was established in Beagle dogs. Bone remodeling was tested at 3, 7, 15, 30, 60 and 90 days after the placement of implants. RANKL and OPG mRNA expression were quantified by real-time PCR. Then mandibular bones were resected and some sections were observed.</p><p><b>RESULTS</b>The most prominent period of bone remodeling occurred at 7 day after the placement of implants (OPG/RANKL mRNA, 2.15 ± 0.1). The expression of RANKL and OPG increased in a time-dependent manner in both soft and hard tissue. After 7 days they gradually decreased.</p><p><b>CONCLUSIONS</b>Both OPG and RANKL were expressed in peri-implant tissues, and the changing tendency of RANKL and OPGmRNA was consistent with the change of bone remodeling. The active stage for bone remodelling in peri-implant tissues during unloading period is about 7 days after implantation.</p>
Subject(s)
Animals , Dogs , Male , Bone Remodeling , Genetics , Dental Implantation , Osteoprotegerin , Genetics , Metabolism , RANK Ligand , Genetics , Metabolism , RNA, Messenger , MetabolismABSTRACT
<p><b>OBJECTIVE</b>To study the effects of combination of angiopoietin-1 (ANG-1) and vascular endothelial growth factor₁₆₅ (VEGF₁₆₅) gene transfer mediated by recombinant adeno-associated viral vector on the neovascularization in chronic ischemic porcine myocardium.</p><p><b>METHODS</b>An ameroid constrictor was implanted around the left circumflex coronary artery (LCX) via endoscopy. Six weeks later, coronary angiography revealed that the myocardial ischemia was established by gradual occlusion of the left circumflex coronary artery (LCX). Sixteen swine with the total occlusion or partial stenosis (> 85 %) of the LCX were divided into 4 groups (4 in each group): group I, group II and group IV (control) received direct myocardium injection of rAAV₂ VEGF₁₆₅, rAAV₂ ANG-1 or PBS alone, respectively; group III received rAAV₂ VEGF₁₆₅ and rAAV₂ ANG-1. Selective coronary angiography and ultrasonography were performed perioperatively to evaluate the cardiac function and the formation of collateral circulation. The expression of VEGF₁₆₅ and ANG-1 proteins were assessed using ELISA or Western blot. The degree of angiogenesis was assessed by use of immunohistochemical analysis.</p><p><b>RESULT</b>Angiography showed that the occlusion of all LCX was completed or exceeded 95% 6 weeks after ameroid constrictor implantation, indicating the successful establishment of animal model. The expression levels of VEGF₁₆₅ in group I and III and ANG-1 in groups II and III began to increase at d7 after transfection and reached the peak at d14; then decreased gradually to the normal level after 3 months. The expression levels of VEGF₁₆₅ in group II and group IV or that of ANG-1 protein in group I and group IV had no markedly changes at different time after transfection. There were significant increase in capillary density and arteriole density and more side branch vessels formed in group III compared with other groups. Echocardiographic measurements showed that the left ventricular systolic function of animals in groups I, II and III increased significantly after gene transfection, especially in group III; but there was no changes in group IV.</p><p><b>CONCLUSION</b>Myocardial perfusion and the left ventricular systolic function are improved after rAAV₂ VEGF₁₆₅ or rAAV₂ ANG-1 transfection, which is associated with the angiogenesis in porcine model of chronic myocardial ischemia.</p>
Subject(s)
Animals , Male , Adenoviridae , Genetics , Angiopoietin-1 , Genetics , Collateral Circulation , Coronary Vessels , Disease Models, Animal , Genetic Therapy , Genetic Vectors , Myocardial Ischemia , Therapeutics , Neovascularization, Physiologic , Swine , Swine, Miniature , Transfection , Vascular Endothelial Growth Factor A , GeneticsABSTRACT
<p><b>OBJECTIVE</b>To create a standard mini-swine model of chronic ischemic myocardium by endoscopy for the research of gene transfer and stem cell.</p><p><b>METHODS</b>Twenty-three male China experimental minipigs were used, aged from 8 to 11 months with a mean of (9.3 +/- 1.8) months and weighed from 20 to 30 kg with a mean of (29.3 +/- 4.3) kg. The myocardial ischemia was established by gradual occlusion of the left circumflex coronary artery (LCX) with an Ameroid constrictor. The Ameroid constrictor was implanted around LCX by endoscopy. Selective coronary angiography, electrocardiogram and Echo-Doppler study were performed perioperatively to evaluate the degree of stenosis.</p><p><b>RESULTS</b>Chronic ischemic myocardial models were successfully generated in 20 of 23 swine by full-endoscopy. Ameroid constrictors were placed at the LCX accurately. Three swine died of anesthetic accident, cardiac arrhythmia at secondary coronary angiography, and pulmonary infection within 6 weeks after operation respectively. Operation time was 25 to 65 min with a mean of (46 +/- 9) min. The blood loss was 30 to 60 ml with a mean of (55 +/- 12) ml. Six weeks later, coronary angiography revealed the total occlusion and partial stenosis (> 85%) of the LCX occurred in 7 and 13 swine respectively. Cardiac systolic and diastolic dysfunction were found in all swine. The ejection fraction value was (65.0 +/- 6.3)% before operation and (41.0 +/- 9.3)% after operation (P = 0.008). The fractional shortening value was (36.2 +/- 4.3)% before operation and (34.2 +/- 2.3)% after operation (P = 0.027).</p><p><b>CONCLUSION</b>The endoscopic surgery is a less invasive way to create a standard mini-swine model of chronic ischemic myocardium with effective results.</p>