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Objective:To study the effects of Zuogui Pills on the expressions of miR-133b-3p and RhoA in osteoclasts of postmenopausal osteoporosis rats; To discuss its potential mechanism.Methods:SD female rats were randomly divided into normal group, model group, sham-operation group, and Zuogui Pills group using a random number table method, with 6 rats in each group. The model group and Zuogui Pills group were treated with oophorectomy to construct a rat model of osteoporosis. Zuogui Pills group was orally administered with Zuogui Pills decoction at a concentration of 10 g/kg for 12 consecutive weeks. Colorimetric method was used to measure the serum calcium and phosphorus levels of rats, and ELISA method was used to detect ALP levels. Bone density meter was used to measure the bone density of the femurs of rats in each group. The osteoclast of each group were cultured, and the expressions of RANKL and RUNX2 protein were detected by Western blot. MiRNA sequencing and differential expression analysis were performed on bone tissues of rats. Osteoclasts were treated with miR-133b-3p mimic and its negative control. The cell proliferation activity of osteoclasts was detected by cell counting kit-8 (CCK-8). The osteoclast differentiation activity was detected by the tartrate-resistant acid phosphatase staining. The dual-luciferase reporter assay was used to detect the relationship between miR-133b-3p and RhoA. The "rescue" experiment of miR-133b-3p mimic and RhoA co-expression were used to study the molecular regulatory mechanism of Zuogui Pills on osteoclast activity.Results:Compared with the model group, the bone mineral density of Zuogui Pills group significantly increased ( P<0.05, P<0.01), the levels of calcium and phosphorus in serum increased, the level of alkaline phosphatase ALP decreased ( P<0.05), the expression of RANKL protein decreased, and the expression of RUNX2 protein increased. Sequencing results showed that rno-miR-133b-3p was down-regulated in osteoclasts of postmenopausa osteoporosis rats treated with Zuogui Pills with the maximum difference ( P<0.01). Q-PCR results showed that the expression of miR-133b-3p in osteoclasts of Zuogui Pills group was significantly lower than that of the model group. The upregulation of miR-133b-3p could significantly promote the cell proliferation and differentiation of osteoclasts. RhoA overexpression could reverse the excessive proliferation and differentiation of osteoclasts caused by miR-133b-3p overexpression. Conclusions:RhoA is the target gene regulated by miR-133b-3p. Zuogui Pills can inhibit the activity of osteoclasts by regulating miR-133b-3p/RhoA axis, relieving the symptoms of osteoporosis.
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@#An innovative approach to quantitatively analyze the histamine and its precursor histidine simultaneously in biological matrices was established for the first time based on double adsorption combined with high performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS).The internal standard was 2-dihydroxybenzoic acid (DHB).The plasma and brain tissue homogenate was protein precipitated with 3-fold acetonitrile, and the supernatant was then sampled for injection analysis.The chromatographic separation of the target components was achieved on an amino chromatography column (ODS-SPXBridge? Amide).Gradient elution was carried out with the mobile phase consisting of solvent A (0.1% formic acid and 1mmol/L ammonium formate in water) and solvent B (acetonitrile).Mass spectrometry was employed for quantitative analysis with ESI ion source in multiple reaction monitoring (MRM) mode.In order to improve the specificity and accuracy, activated carbon and calcite were used for the double adsorption of biological matrices for the first time.The adsorbed matrix was then used for methodology validation.The results showed that histamine and histidine were linear in the quantitative range (correlation coefficient r ≥ 0.999).Accuracy, precision, extraction recovery, matrix effect and stability all met the requirements of biological sample analysis.All results suggested that the present method could not only be efficiently and reliably used for simultaneous quantitative analysis of histamine and histidine in biological samples, but also provide reference for the detection of other endogenous substances.
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BACKGROUND: Children and adolescents are in the growth and development stage, so the use of the same reconstruction methods as adults is easy to induce complications including limb length differences, high graft failure rate and re-surgical intervention. Simultaneously, the occurrence of osteoarthritis will also be advanced. OBJECTIVE: To explore the anatomical characteristics of the anterior cruciate ligament between children, adolescents and adults on MRI, and to provide an anatomical basis for the reconstruction of the anterior cruciate ligament in children and adolescents. METHODS: A retrospective analysis of patients undergoing knee MRI examination in Qingdao Municipal Hospital from October 2016 to October 2018 was conducted. These patients were divided into child and adolescent group and adult group (n=48 per group). The angle between the anterior cruciate ligament and the tibia and femur on the sagittal plane and the position of the anterior cruciate ligament tibial insertion, the angle between the anterior cruciate ligament and the tibia on the coronal plane, the tibial insertion of the anterior cruciate ligament and femur, and the position of the femur stop point on the axial position were measured in both groups. The data obtained from the measurement of the anterior cruciate ligament of children and adolescents were processed. The growth curves of the shape and position of the anterior cruciate ligament of children and adolescents were drawn, and the law of its growth changes was analyzed. RESULTS AND CONCLUSION: (1) The angle between the sagittal plane of the anterior cruciate ligament and the femur (t=﹣2.906, P<0.05), the angle between the sagittal plane of the anterior cruciate ligament and the tibia (t=﹣10.280, P < 0.05), the anterior cruciate ligament angle between the coronal plane and the tibia (t=﹣5.714, P<0.05) were smaller in the child and adolescent group than those of the adult group, and the difference was significant. (2) The ratio of the tibia coronal plane of the anterior cruciate ligament (t=﹣7.263, P < 0.05) and the ratio of the anterior cruciate ligament and femur axial plane (t=﹣7.378, P < 0.05) were lower in the child and adolescent group than those of the adult group, and the difference was significant. (3) There was no significant difference in the anterior cruciate ligament and tibia sagittal plane ratio (t=﹣1.588, P>0.05) and anterior cruciate ligament and femoral coronal surface ratio (t=﹣1.647, P>0.05) between the child and adolescent group and the adult group. (4) The growth curve results showed that during the growth and development, the angle between the anterior cruciate ligament and the femur and tibia on the sagittal plane and the angle with the tibia on the coronal plane changed from small to large (P<0.05). The relative position of the tibial insertion in the coronal position changed from small to large, which indicates that during the growth and development, the tibial insertion moved from inside to outside relative to the inside of the tibial plateau on the coronal plane (P < 0.05). (5) The relative position of femoral insertion in the axial position changed from small to large, suggesting that during the growth and development, the femoral insertion moved from the outside to the inside relative to the lateral femoral condyle (P<0.05). (6) There was no significant difference in the tibial insertion and the femoral insertion between the child and adolescent group and adult group (P>0.05).
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Objective:To investigate the clinical efficacy of double tract reconstruction of proximal gastric resection and Roux-en-Y total gastrectomy in the treatment of adenocarcinoma of the esophagogastric junction, in order to provide reference for clinical selection.Methods:From June 2016 to June 2018, patients with adenocarcinoma of the esophagogastric junction in Chifeng City hospital were prospectively selected and randomly divided into proximal gastrectomy double channel anastomosis group (PG group) and total gastrectomy Roux-en-Y anastomosis group (TG group). The perioperative indexes, clinical pathology, early (≤ 30 days) and late (> 30 days) complications of the two groups were compared, and the nutritional status [body mass index (BMI), prealbumin, albumin, total protein], and anemia degree [hemoglobin (Hb), red blood cell count (RBC)] of the two groups at 3 months, 6 months and 12 months after operation were compared.Results:A total of 83 patients with adenocarcinoma of the esophagogastric junction were admitted, including 62 patients who were eligible for surgery, 31 cases in each group, 1 case in each group was lost to follow-up, and 30 cases in each group. There were no tumor recurrence or death in two groups. There were no statistically significant differences in age, gender, tumor size, tumor stage, preoperative complications, preoperative nutritional status and anemia between the two groups ( P>0.05). The time of first exhaust, time of starting fluid feeding and postoperative hospitalization in PG group was significantly earlier than that in TG group [(3.0±0.7)days vs (4.2±0.7)days, (7.9±0.9)days vs (9.0±0.9)days, (13.3±1.1)days vs (14.6±0.9)days, P<0.05], while the operation time was slightly longer [(252.0±36.9)min vs (225.8±31.1)min, P<0.05]. There was no significant difference in intraoperative blood loss and number of lymph node dissection between PG group and TG group [(241.7±81.0)ml vs (245.8±71.9)ml, (40.5±12.2)nodes vs (43.2±10.3)nodes, P>0.05]. There was no statistically significant difference in the occurrence of early and late postoperative complications ( P>0.05). The incidence of reflux esophagitis in PG group was less than that in TG group ( P<0.05). The nutritional status of PG group was significantly better than TG group in 3 months, 6 months and 12 months after the operation ( P<0.05). The Hb and RBC decreased in both groups at 3 and 6 months after operation, and the Hb and RBC in PG group were higher than those in TG group. There was no anemia in PG group and TG group 12 months after operation. Conclusions:Double tract reconstruction of proximal gastric resection is effective in the treatment of adenocarcinoma of the esophagogastric junction. It is worthy of further clinical promotion.
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<p><b>OBJECTIVE</b>To investigate the infiltration and prognostic significance of tumor-infiltrating mast cells (TIMs) in nasopharyngeal carcinoma (NPC).</p><p><b>METHODS</b>Immunohistochemistry for tryptase was performed on 154 NPC specimens. The median value of TIM density was used as a cutoff point to separate the patient cohort into two groups with either low or high TIM infiltration. The associations between TIM and clinicopathological factors were analyzed by Mann-Whitney U text. Survival curves were plotted according to the Kaplan-Meier method. Univariate and multivariate analyses were performed using the Log-rank test and the Cox proportional hazard models, respectively. P<0.05 was considered statistically significant. All statistical analyses were conducted using SPSS 13.0.</p><p><b>RESULTS</b>TIM was mainly in the stroma of NPC and detected in all specimens. The median value of TIM density (25.60/high power field) was used as a cutoff point to separate the patient cohort into two groups with either low or high TIM infiltration. The density of TIM was positively correlated with N stage (Z=-2.193, P<0.05) and clinical stage (Z=-2.551, P<0.05). The 3-year overall survival (OS) and progression-free survival (PFS) of patients were 64.4% and 55.7% in the high TIM density group; 78.3% and 77.0% in the low TIM density group. For survival evaluation, high density of TIM was associated with worse OS and PFS (P<0.05). Multivariate Cox regression model analysis showed TIM infiltration was an independent risk factor for both OS and PFS.</p><p><b>CONCLUSIONS</b>The density of TIM in NPC increased with tumor stage. High TIM infiltration was associated with poor overall survival and progression-free survival.</p>