ABSTRACT
ObjectiveFrom the perspective of energy metabolism， the mechanism of Osteoking （OK） in the treatment of myofascial pain syndrome （MPS） was revealed through systems biology prediction combined with holistic animal experimental validation methods. MethodFirstly， the key targets of MPS and their related molecular mechanisms were predicted by the systems biology method， and the core network targets were screened. Then， the network-predicted targets were verified by animal experiments. Specifically， 60 SD rats were randomly divided into normal group， model group， low， medium， and high dose OK groups （0.66， 1.31， 2.63 mL·kg-1）， and positive celecoxib group （21 mg·kg-1）. The MPS model was established by beating combined with a centrifugal exercise method for eight weeks. Except for two days after modeling， the intervention of OK or celecoxib was performed. After the completion of the model， the drug was administered for two weeks. The histopathological changes of trigger point muscle tissue were observed by hematoxylin-eosin staining. The content/activity of Na-K-ATP enzyme （Na+-K+-ATPase）， Ca2+ pump （Ca2+ATPase）， Ca2+， lactate dehydrogenase （LDH）， glutathione （GSH）， malondialal （MDA）， superoxide dismutase （SOD）， cyclic adenosine phosphate （cAMP）， and protein kinase A （PKA） in serum and/or trigger point muscle tissue in MPS rats was detected by enzyme-linked immunosorbent assay. Protein expression levels of PKA and the peroxisome proliferator-activated receptor γ coactivator 1α （PGC1α） in MPS rats were detected by immunohistochemistry. The protein expression levels of PKA， PGC1α， and mitochondrial transcription factor A （TFAM） in MPS rats were detected by Western blot. ResultThe network prediction results suggest that OK acts on the key target of energy metabolism related to the occurrence and development of MPS and may participate in the activation of the cAMP/PKA/PGC1α signaling pathway. The experimental validation results show that compared with the normal group， contracture nodules and disordered arrangement of muscle fibers appear in the trigger point muscle tissue of MPS rats. Na+-K+-ATPase， Ca2+ATPase， SOD activity， Ca2+， and GSH contents in serum and/or trigger point muscle tissue are significantly decreased （P<0.01）. Both LDH activity and MDA contents are significantly increased （P<0.01）， and the protein expression levels of cAMP， PKA， PGC1α， and TFAM are significantly decreased （P<0.01）. Compared with the model group， OK improves the histopathological morphology of trigger point muscle fibers in MPS rats， and after the intervention of OK， Na+-K+-ATPase， Ca2+ATPase， SOD activity， Ca2+， and GSH contents in serum and/or trigger point muscle tissue in MPS rats are significantly increased （P<0.05， P<0.01）. LDH activity and MDA contents are significantly reduced （P<0.05， P<0.01）. The protein expression levels of cAMP， PKA， PGC1α， and TFAM are significantly increased （P<0.05， P<0.01）. ConclusionThe mechanism of OK's intervention in MPS rats may be related to its effective activation of the cAMP/PKA/PGC1α signaling pathway， thus promoting mitochondrial energy metabolism and trigger point muscle fiber damage repair in muscle cells.
ABSTRACT
Objective:To evaluate the data quality of Shenzhen Type 1 Diabetes Alliance (SZT1D), and to provide a basis for evaluation and improvement for the continuous improvement of data quality.Methods:From December 2018 to July 2021, 697 first-visit type 1 diabetes (T1DM) patients (including 501 in Shenzhen and 196 out-of-Shenzhen) and 120 re-visited T1DM patients (including 113 in Shenzhen and 7 out-of-Shenzhen) who were registered by SZT1D in collaborative research platform network of China Type 1 Diabetes Alliance (hereinafter referred to as China T1D). The data quality was evaluated from three dimensions: data completion, accuracy and revisit. The data completion degree was evaluated by the overall data completion degree and the key indicator completion degree; the data accuracy was evaluated by the probability of abnormal blood glucose value; the patient′s return visit was evaluated by the return visit rate.Results:The main characteristics of T1DM in SZT1D were young and middle-aged adults [age: (34.4±17.1)years] with thin body [BMI: (19.80±3.52)kg/m 2)], half of male and female patients [proportion of male: 52.4%(365/697)]; the main types of diagnosis were classical T1DM [65.22%(150/230)] and latent autoimmune diabetes in adults(LADA) [26.08%(60/230)], and the fasting blood glucose (FPG) [(10.93±6.98)mmol/L] and glycosylated hemoglobin (HbA 1c) [(10.63±3.01)%] were high. The average completion rate of the overall data of the first diagnosed patients in SZT1D was only 60% [(62.9±31.5)%]: the number of patients with overall data completion ≥80% in SZT1D was only 50.2%(350/697); the number of patients with overall data completion ≥80% in Shenzhen was less than that outside Shenzhen [44.3%(222/501) vs 65.3%(128/196), P<0.001]. The key indicators with better completion rate of first-visit were disease course [76.2%(531/697)], age of onset [75.8%(528/697)], family history of diabetes [74.9%(522/697)], etc., but none of them had a completion rate of more than 80%, and the diabetes self-management behavior assessment questionnaire and scale score were completely missing; the frequency of daily blood glucose monitoring [46.1%(231/501) vs 64.3%(126/196), P<0.001], current insulin regimen [44.3%(222/501) vs 63.3%(124/196), P<0.001], number of diabetic ketoacidosis (DKA) since the onset of the disease [45.7%(229/501) vs 64.8%(127/196), P<0.001] and the number of symptomatic hypoglycemia in the past 1 month [39.3%(197/501) vs 63.8%(125/196), P<0.001] were higher in Shenzhen than those reported outside Shenzhen. In addition, the probability of abnormal FPG and postprandial glucose (PPG) [5.2%(24/466); 3.8%(19/236)] were low. The revisit rate was not high [17.2%(120/697)], and the revisit rate in Shenzhen was higher than that outside Shenzhen [22.6%(113/501) vs 3.6%(7/196), P<0.001]. The first revisit rate was 16.2%(113/697) and the second revisit rate was seriously insufficient [1.0%(7/697)]. Conclusions:The data quality of T1DM patients recorded by SZT1D needs to be further improved. Improving the information interconnection between China-T1D and SZT1D, employing quality control personnel and building a systematic data quality evaluation analysis and feedback mechanism are methods to promote the comprehensive, accurate and efficient input of T1DM data and continuously improve the evaluation methods to improve the overall data quality.
ABSTRACT
Objective To investigate the possibility of the involvement of RhoA/mDia1 pathway to cause the expression of phosphorylate ezrin-radixin-moesin (p-ERM) in rat pulmonary micro-vascular endothelial cell (PMVEC) after the stimulation of lipopolysaccharide (LPS).Methods The specimens of lung tissue were taken from healthy male SPF grade SD rat with 100-120 g body weight which was purchased from the laboratory animal center of Anhui province.After culture,the PMVECs were randomly divided into dose-dependent groups (0,0.1,1,10 μg/mL LPS added in PMVECs and cultured for30 min,n =8 in each),time-dependent groups (10 μg/mL LPS added to PMVECs cultured for 0,15,30,60,120 min,n =8 in each) and intervention group (n =8).In the intervention group,PMVECs were cultured with 1 μg/mL C3 transferase in serum free media for 240 min,followed by treatment with 10 μg/mL LPS for 30 min.Meanwhile,two control groups in serum-free DMEM medium were made by adding 10.μg/mL LPS to PMVECs and 1 μg/mL C3 transferase to PMVECs respectively cultured for 30 min (n =8 in each).Western blot was used to detect the level of p-ERM,ERM and mDia1.Data were analyzed with SPSS 16.0 software,while one way analysis of variance (ANOVA) was used to compare multiple sets of variables,the intergroup comparisons were analyzed by the least-significant-difference (LSD) tests,with P ＜0.05 for the statistically significant difference.Results ERM,p-ERM and mDia1 were presented in rat PMVEC.Stimulation with LPS up-regulated p-ERM,mDia1 in a dose-dependent manner:LPS [0 μg/mL LPS group:(0.520±0.101),0.1 μg/mL LPS group:(0.657 ±0.092),1 μg/mL LPS group:(0.891 ±0.167),10 μg/mL LPS group:(1.227 ±0.106);0 μg/mL group vs.0.1 μg/mL group,P ＞0.05;the rest P ＜0.01];and mDia1 [0 μg/mL LPS group:(0.200 ±0.102),0.1 μg/mL LPS group:(0.430 ±0.121),1 μg/mL LPS group:(0.603 ±0.154),10 μg/mL LPS group:(0.887 ±0.204);0.1 μg/mL group vs.1 μg/mL group,P ＞ 0.05;the rest P ＜ 0.05].In time-dependent group,the level of p-ERM protein increased at 15 min (0.670 ±0.149),peaked at 30 min (1.175 ±0.103),then decreased,at 60 min (0.959 ±0.189),90 min (0.842 ±0.129),but kept at higher level at 120 min (0.767 ±0.097) than that in control group (0.471 ±0.157,15 min group vs.120 min group,60 min group vs.90 min group and 90 min group vs.120 min group,P ＞ 0.05;the rest P ＜ 0.05);and the level of mDia1 increased at 15 min (0.779 ±0.035),peaked at 30 min (0.889 ±0.036) then decreased at 60 min (0.648 ±0.019),90 min (0.582 ±0.068),but kept at higher level at 120 min (0.526 ±0.059) than that in control group (0.284±0.118,all P ＜ 0.01).C3 transferase caused a marked attenuation of LPS induced p-ERM expression [control group:(0.339 ± 0.069);C3 + LPS group:(0.438 ± 0.07);C3 control group:(0.352 ± 0.071);LPS control group:(0.634 ± 0.191),C3 + LPS group vs.LPS control group,P =0.01],as the same in mDia1 [control group:(0.449 ±0.122);C3 + LPS group:(0.380 ±0.148);C3 control group:(0.404 ±0.164);LPS control group:(0.622 ±0.174),C3 + LPS group vs.LPS control group,P ＜ 0.01].Conclusions LPS could up-regulated the expression of p-ERM protein,and inhibition of RhoA/mDia1 signal pathway by C3 transferase could down-regulated the p-ERM levels.
ABSTRACT
Objective To analyze serum 25(OH)D level and the influence factors in preterm infants. Methods The data of serum 25(OH)D level in preterm infants born from July 2012 to June 2014 were retrospectively analyzed along with gestational age, gender, parity, delivery mode, birth season, birth weight, age of the mother and pregnancy complications. Results Totally 325 cases were included and their average gestational age was 33.41±2.29 week, the ratio of male to female was 1.32?1 and average serum 25(OH)D level was 37.34±16.17 nmol/L. The incidence of vitamin D deficiency and insufficiency in preterm infants was 37.8% and 41.8% respectively. Serum 25 (OH) D levels in preterm infants born in summer and autumn were higher than those born in winter and spring, and there was statistical difference (P<0.05). Logistic regression analysis showed that birth season and the mother's age may be the risk factors that influence serum 25 (OH) D levels in preterm infants. Conclusion The incidence of vitamin D deficiency and insufficiency in preterm infants is high, and the factors affecting the level of vitamin D need to be further studied.
ABSTRACT
Purpose To investigate the effects of chemotactic factor CCR4 on the abi1ity of pro1iferation,ce11 cyc1e,invasion,and mi-gration of human ga11b1adder cancer ce11. Methods Western b1ot was used to detect the expression 1eve1 of CCR4 in ga11b1adder carci-noma ce11s. Ga11b1adder carcinoma ce11s was infected by means of s1ow virus,the CCR4 gene si1encing was conducted using siRNA-CCR4 interference techno1ogy. Ga11b1adder carcinoma ce11s GBC-SD were divided into three groups( GBC-SD,GBC-SD/CCR4-RNAi and GBC-SD/contro1). CCL17,a 1igand of CCR4,was used to act on these three groups of ce11s. CCK8 method was used to detect the ce11 pro1iferation abi1ity of three groups. F1ow cytometry was used to test ce11 cyc1e. Tanswe11 assay was app1ied to detect ce11 migration and invasion abi1ity. Western b1ot was performed to detect the expression of its corresponding 1igands CCL17 and CCL22 proteins. Re-sults CCR4 gene si1ence did not inf1uence ce11 cyc1e and pro1iferation of ga11b1adder ce11 GBC-SD,but can significant1y inhibit GBC-SD ce11 invasion and movement abi1ity,CCR4 gene si1ence had no inf1uence on the expression of CCL17 and CCL22 gene in tumor ce11s. Conclusion Ga11b1adder carcinoma ce11s GBC-SD express chemokine receptor CCR4,chemokine receptor CCR4 can promote the invasion and metastasis of GBC-SD ce11s.
ABSTRACT
Objective:To compare the amino acid sequences difference of HA,NA novel influenza virus A/H7N9 isolates, decipher possible B cell epitopes and T cell epitopes of HA,NA protein,and analyze the association between susceptibility and HLA polymorphisms.Methods:The amino acid sequences of novel influenza A ( H7N9) virus were downloaded from Genbank.Phylogenetic trees were constructed based on the amino acid sequences of HA and NA by using software Clustal X and MEGA 4.0.B cell and T cell epitopes were respectively predicted with Protean software and NetMHCⅡ2.2 Server online server.Results:The homology of HA and NA proteins of H7N9 virus was high.10 B cell epitopes and 15 T cell epitopes were randomly distributed throughout HA sequence and 12 B cell epitopes and 9 T cell epitopes were randomly distributed throughout NA sequence.HLA-DRB1*0701 allele which was commonly observed in Northern Chinese population have a high binding affinity for 9-mer peptides of HA and NA proteins.Conclusion:The prediction of B and T cell epitopes of HA and NA proteins with multiple methods benefits the research and development of vaccine against human infection with avian influenza A H7N9 virus.HLA-DRB1*0701 allele may contribute to susceptibility to novel influenza A (H7N9) virus.H7N9 influenza virus is more easily spread in Urumqi,Harbin,Shandong Province,Liaoning Province,Beijing, Shijiazhuang and Tianjin of China.