BACKGROUND: Hippocampal neuron presents remarkably injury in cerebral after seizure of epilepsy. Necrosis and apoptosis are two kinds of neural cell injury after epilepsy and play an important role in neural injury of epilepsy. Being endogenous neural protective transmitter, adenosine may inhibit the release of excitatory amino acid, production of oxygenic free radical and action of nitric oxide. Simultaneously, it can improve cerebral blood flow and anti-convulsion. But it has been unknown concerning to the relationship between adenosine and cell apoptosis after epilepsy yet.OBJECTIVE: To observe the effects of 2-CAdo adenosine-receptor excitant on genetic expression of bcl-2, Bax of hippocampal cells in epileptic rats and further probe into the mechanism of adenosine on anti-convulsion and brain protection.DESIGN: Completely randomized controlled experimental research in which the experimental animals were taken as the objects.SETTING: Pediatrics department and general surgical department of one oil field general hospital, and pediatric internal department of a hospital affiliated to one university.MATERIALS: The experiment was performed in Experimental Zoology Departnent and Pathological Teaching & Research Department of Harbin Medical University from October 2002 to March 2003. Totally 104 Wistar rats of either sex were employed, weighing varied from 200 g to 250 g. The animals were randomly divided, named as normal group 8 rats, epileptic group 32 rats, epileptic & 2-CAdo group 32 rats, and epileptic & physiological saline group 32 rats.INTERVENTIONS: The animal epileptic model was set up by intra-abdominal injection of coriamyrtin 15 mg/kg(provided by Pathology Department of Harbin Medical University. Convulsion presented in all of rats, 5 minutes later after injection, lasting for 1 or 2 minutes. In epileptic & 2-CAdo group, 2-CAdo(provided by ICN company), 0.6 mg/kg, was injected from the vein on the tail 1 hour before coriamyrtin injection and 1 hour after convulsion respectively. In epileptic & physiological saline group, the physiological saline of equal dosage was injected from the vein on the tail 1 hour before coriamyrtin injection and 1 hour after convulsion respectively.MAIN OUTCOME MEASURES: Positive cell counts of bcl-2 and Bax genetic expression in hippocampal CA1 area.RESULTS: Twenty-four hours after epilepsy seizure, neural cell bcl-2 expression was increased in hippocampal CA1 area, was remarkably decreased in 48 hours, and the expression was only little amount in 72 hours, but it was increased again in 7 days. Bax expression began increased in 24 hours after epilepsy seizure, was significantly increased in 48 hours, reached the peak in 72 hours, the expression was the minimum in 7 days. In epileptic & 2-CAdo group, bcl-2 expressions at corresponding times were remarkably increased compared with epileptic group and epileptic & physiological saline group( P＜ 0.05), Bax expressions were remarkably decreased compared with epileptic group and epileptic & physiological saline group( P ＜ 0.05), indicating statistical significance.CONCLUSION: 2-CAdo can reduce apoptosis of hippoeampal neural cells after epilepsy seizure and provide a certain protection for neural cells.

Objective To study the drug dosage and time dependency characteristics of high-fructose-high-fat-feeding plus dexamethason for inducing the mouse acute fatty liver model and to optimize the condition of drug induced fatty liver model.Methods Male KM mice were divided into the normal control group and high-fructose-high-fat-feeding plus peritoneal injection of dexamethason group.The mice were killed at 3 different time points.The mouse body mass and liver mass were detected.The liver index was calculated.The serum and liver tissue homogenate TG and serum glucose(GLU) levels were detected.The liver tissue pathological change was observed by HE staining.Total RNA reverse expression related gene was extracted from the liver tissue.The total protein was extracted from the liver tissue and the related protein expression was detected by Western Blot.Results Compared with the control group,blood and liver homogenate TG after 7 d in the dexamethason model group was increased,the liver index was increased,the pathological section displayed that the fatty liver was formed.RT-PCR showed that lipid metabolism related gene expression had obvious change.Western Blot showed that SIRT1 was significantly decreased.But with the dexamethason dosage decrease and time extending,the fatty liver related indexes were decreased,lipid metabolic gene PPAR,FOXO3 and FXR were gradually increased,while LXR was gradually decreased and protein SIRT1 was gradually increased.Conclusion High-fructose-high-fatfeeding plus peritoneal injection of dexamethason could establish the mouse acute fatty liver model,moreover the model maintenance has dependency on dexamethason dosage and medication time,which has a guidance significance for the drug interventional experiment.

Objective:To verify the determination method of iodine in serum by inductively coupled plasma mass spectrometry (ICP-MS) and to evaluate the consistency between ICP-MS and As 3+-Ce 4+ catalytic spectrophotometry in determination of serum iodine. Methods:Serum iodine concentration was determined by ICP-MS, 187Re was used as an internal standard, and ralated parameters were optimized. Eighty-eight serum samples were simultaneously determined by ICP-MS and As 3+-Ce 4+ catalytic spectrophotometry, and the evaluation indexes included determination range of standard curve, detection limit, precision, accuracy. In addition, we also evaluated the consistency of the two methods through inter-group correlation analysis, intra-group correlation coefficient analysis, Passing-Bablok regression and Bland-Altman analysis. Results:The linear range of ICP-MS standard curve was 0 - 300 μg/L. There was a good linear correlation between iodine concentration value and iodine response value, and the correlation coefficient range was 0.999 8 to 0.999 9. The detection limit of the ICP-MS method was 1.96 μg/L. The relative standard deviation ( RSD) ranged from 0.2% to 1.4% and from 0.4% to 1.8% for intra and inter-batch precision tests of serum samples. The recovery rate ranged from 90.44% to 108.71%. The correlation analysis of 88 serum samples showed that there was a good correlation between the two methods ( r = 0.934, P < 0.05), and the intra-class correlation coefficient was 0.932. The results of Passing-Bablok regression showed that there was no significant difference between the two methods ( P > 0.05). Bland-Altman diagram suggested that the results of the two methods were consistent. Conclusions:ICP-MS method has low detection limit, high precision and accuracy. ICP-MS method is simple, rapid, easy and suitable for determination of iodine in large quantities of serum samples. The results of the two methods for determining serum iodine are consistent.

Bone marrow-derived mesenchymal stem cells (BMSCs) for repairing damaged heart tissue are a new kind of important treatment options because of their potential to differentiate into cardiomyocytes. We in this experiment investigated the effect of different electrical stimulation time on the expression of myocardial specificity gene and protein in rat bone marrow mesenchymal stem cells (rBMSCs) in vitro. The rBMSCs of second or third generation were randomly divided into three groups, i.e, electrical stimulation (ES) group, 5-Azacytidine (5-Aza) group and the control group. The rBMSCs in the ES groups with complete medium were exposed to 2 V, 2 Hz, 5 ms electrical stimulation for 0. 5 h, 2 h, 4 h, and 6 h respectively every day for 10 days. Those in the 5-Aza group were induced by 5-Aza (10 μmol/L) for 24 h, and then cultured with complete medium for 10 days. Those in the control group were only cultured with complete medium, without any treatment, for 10 days. The rBMSCs' morphological feature in each group was observed with inverted phase microscope. The mRNA expression of myocyte-specific enhancer factor 2C (MEF-2C) and connexin 43 (Cx43) were examined with Real-Time quantitative PCR and the protein expression of MEF-2C, Cx43 were detected with Western Blot method. The results showed that the mRNA expression level of the MEF-2C, Cx43 and the protein expression level of MEF-2C, Cx43 were significantly higher in the ES group and 5-Aza group than those in the relative control group (P < 0.05). It suggests that electrical stimulation could play a part of role in the induction of the rBMSCs to differentiate into the cariomyocyte-like cells in vitro and the effectiveness of the electrical stimulation with 2 h/d had the best in our experiment. But the mechanism how electrical stimulation promotes the differentiation of rBMSC into cardiomyocyte is still unclear.
Animals ; Biomarkers ; metabolism ; Cell Differentiation ; Cells, Cultured ; Connexin 43 ; metabolism ; Electric Stimulation ; MEF2 Transcription Factors ; metabolism ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Myocytes, Cardiac ; cytology ; RNA, Messenger ; metabolism ; Rats ; Rats, Sprague-Dawley

The in vitro release is an important index to evaluate the quality of liposome formulation.Currently, there is no evaluation method for the in vitro release of liposome in pharmacopoeia of various countries, which leads to the lack of unified standard and safety guarantee for the quality evaluation of liposome formulation.Taking the self-made paclitaxel derivative liposomes as an example, the paddle membrane binding method established by optimizing external release conditions was used to simulate the complete release of paclitaxel derivative drugs in 12 hours under physiological conditions.The results showed that using 0.5% SDS-HEPES as the release medium and a dialysis bag with a molecular weight cutoff of 1 000 kD to release the liposome solution met the requirements and had discrimination ability, providing a reference for the development of drug-loaded liposomes release methods in vitro.

Objective:To summarize the concept, theoretical basis, evaluation tools and mechanisms, influencing factors, and intervention measures of kinesiophobia.Methods:The literature on kinesiophobia in patients undergoing total knee replacement was electronically searched on databases such as China National Knowledge Infrastructure, China Biology Medicine disc, WanFang Data, PubMed, CINAHL, Web of Science, Embase, Scopus, PsycINFO, Cochrane Library. The search period was from database establishment to June 24, 2023. This study extracted and analyzed data from the included literature.Results:A total of 32 articles were included. The Tampa Scale for Kinesiophobia was a widely used tool for evaluating kinesiophobia. The influencing factors of kinesiophobia were demographic and disease factors, body motor function, and psychological and social factors. The intervention measures for kinesiophobia mainly included cognitive behavioral intervention, pain health education, exercise, art video or music intervention, multidisciplinary collaborative intervention, and so on.Conclusions:The concept and theoretical basis of kinesiophobia are not yet complete. It is necessary to revise and improve the theoretical model and assessment tool for kinesiophobia and construct an intervention program for kinesiophobia in combination with the concept of rapid rehabilitation.

BACKGROUND:Successful uterine spiral artery remodeling is necessary for normal pregnancy,in which vascular smooth muscle cells are important cells.Interferon-γ is associated with the loss of vascular smooth muscle cells during early pregnancy.However,the specific mechanism is not fully understood. OBJECTIVE:To investigate the effects of interferon-γ on migration and apoptosis of vascular smooth muscle cells through NLRP3/caspase-1/GSDMD pyroptosis pathway. METHODS:Human vascular smooth muscle cells were divided into control group and interferon-γ group.The control group was cultured normally,and the interferon-γ group was treated with 10 ng/mL interferon-γ for 24 hours.The migration ability of vascular smooth muscle cells was detected by Transwell assay.The apoptosis of vascular smooth muscle cells was detected by TUNEL assay and flow cytometry.The mRNA expression levels of NLRP3 and caspase-1 were detected by qPCR.Western blot assay was utilized to detect NLRP3,caspase-1,and cleaved N-terminal GSDMD protein expression levels. RESULTS AND CONCLUSION:Compared with the control group,the migration ability and apoptosis rate of vascular smooth muscle cells in interferon-γ group were significantly increased(P<0.05).Compared with the control group,the mRNA expression levels of NLRP3 and caspase-1 in vascular smooth muscle cells of interferon-γ group were significantly increased(P<0.05).Compared with control group,the expression levels of NLRP3,caspase-1,and cleaved N-terminal GSDMD protein in vascular smooth muscle cells in the interferon-γ group were significantly increased(P<0.05).The results suggest that interferon-γ may regulate the migration and apoptosis of vascular smooth muscle cells through NLRP3/caspase-1/GSDMD pyroptosis pathway.

Objective To analyze the expressions of lymphocytes in the peripheral blood and serum cytokines in the children with secretory otitis media (SOM) and the effects of glucocorticoids interventions.Methods Totally 90 SOM children were selected as case group,and 30 healthy children were selected as control group.The case group was randomly divided into group A (simple oral antibiotic treatment),group B (oral antibiotics combined with local glucocorticoid treatment) and group C (oral antibiotics combined with systemic glucocorticoid treatment),30 cases in each group.The CD4 + T lymphocytes percentage,CD8 + T lymphocytes percentage,the ratio of CD4 +/CD8 + T lymphocytes in peripheral blood and the serum interleukin-2 (IL-2),interferon gamma (IFN-γ),tumor necrosis factor alpha (TNF-α),interleukin-6 (IL-6),interleukin-10 (IL-10) levels were detected and compared between case group and control group.The air conduction auditory thresholds under different frequency of the patients in group A,group B and group C were examined and compared.Results The CD4 + T lymphocytes percentage,the ratio of CD4 +/CD8 + T lymphocytes in peripheral blood and the serum IL-2,IFN-γ,TNF-α,IL-6,IL-10 levels of the patients in the case group were significantly higher than those in the control group (P ＜ 0.0 5).After the treatment,the above indicators of the children in group A,group B and group C decreased,and there were significant differences before and after the treatment between two groups (P ＜ 0.05).The air conduction auditory thresholds under different frequency of the patients in group B or group C were significantly lower than those in group A (P ＜ 0.05),while there were no significant differences in the air conduction auditory thresholds under different frequency between the children in group B and group C (P ＞ 0.05).Conclusion The patients with SOM show imbalanced cell immune function and cytokines expressions in the peripheral blood.The combination of glucocorticoids therapy and routine antibiotic therapy can effectively improve the immune function and the therapeutic effects.

Objective To analyze the expressions of lymphocytes in the peripheral blood and serum cytokines in the children with secretory otitis media (SOM) and the effects of glucocorticoids interventions.Methods Totally 90 SOM children were selected as case group,and 30 healthy children were selected as control group.The case group was randomly divided into group A (simple oral antibiotic treatment),group B (oral antibiotics combined with local glucocorticoid treatment) and group C (oral antibiotics combined with systemic glucocorticoid treatment),30 cases in each group.The CD4 + T lymphocytes percentage,CD8 + T lymphocytes percentage,the ratio of CD4 +/CD8 + T lymphocytes in peripheral blood and the serum interleukin-2 (IL-2),interferon gamma (IFN-γ),tumor necrosis factor alpha (TNF-α),interleukin-6 (IL-6),interleukin-10 (IL-10) levels were detected and compared between case group and control group.The air conduction auditory thresholds under different frequency of the patients in group A,group B and group C were examined and compared.Results The CD4 + T lymphocytes percentage,the ratio of CD4 +/CD8 + T lymphocytes in peripheral blood and the serum IL-2,IFN-γ,TNF-α,IL-6,IL-10 levels of the patients in the case group were significantly higher than those in the control group (P ＜ 0.0 5).After the treatment,the above indicators of the children in group A,group B and group C decreased,and there were significant differences before and after the treatment between two groups (P ＜ 0.05).The air conduction auditory thresholds under different frequency of the patients in group B or group C were significantly lower than those in group A (P ＜ 0.05),while there were no significant differences in the air conduction auditory thresholds under different frequency between the children in group B and group C (P ＞ 0.05).Conclusion The patients with SOM show imbalanced cell immune function and cytokines expressions in the peripheral blood.The combination of glucocorticoids therapy and routine antibiotic therapy can effectively improve the immune function and the therapeutic effects.

Objective:To study the clinical characteristics of growth development and metabolic disorders in obese children and adolescents with insulin resistance (IR).Methods:Normal weight or obese children and adolescents who hospitalized at the Department of Children′s Health Care of Children′s Hospital Affiliated to Nanjing Medical University from September 2015 to April 2018 were recruited.Children′s height, body weight and waist circumference were measured, and waist-to-height ratio (WHtR) and body mass index (BMI) were calculated.Puberty process was determined by Tanner stage.Blood glucose, blood lipid and insulin were measured in fasting state, and homeostasis model assessment of insulin resistance (HOMA-IR) was calculated based on fasting blood glucose and insulin levels.IR was considered when HOMA-IR was over 2.69.Non-alcoholic fatty liver (NAFLD) was diagnosed by abdominal ultrasound.Results:(1) A total of 691 subjects were included, including 183 cases with the age of (9.73±2.38) years in the normal weight group/normal group, and 508 cases with the age of (10.24±2.05) years old in the obese group.The rate of IR was higher in obese group than that in normal group (55.71% vs. 10.38%), and the difference was statistically significant ( P<0.001). (2)HOMA-IR was positively correlated with age ( r=0.256, P<0.001), BMI ( r=0.426, P<0.001), waist circumference ( r=0.454, P<0.001), and WHtR ( r=0.321, P<0.001). After the adjustment for age, sex, and puberty stage, HOMA-IR was also positively correlated with BMI ( r=0.418, P<0.001), waist circumference ( r=0.419, P<0.001) and WHtR ( r=0.375, P<0.001). (3) During puberty, HOMA-IR in both of obese group and normal group was increased, and HOMA-IR in obese group was more particularly serious compared to normal group[TannerⅠ: 2.60(1.49, 3.94) vs.1.28(0.80, 1.90); Tanner Ⅱ: 3.07(1.75, 5.17) vs.1.80(1.16, 2.96); Tanner Ⅲ: 4.33(2.80, 6.57) vs.2.47(1.41, 3.68); Tanner Ⅳ-Ⅴ: 3.49(1.04, 5.78) vs.1.91(0.54, 2.60)], and the differences were all statistically significant(all P<0.05). (4)Compared with the obese objects without IR, obese children and adolescents with IR had higher systolic blood pressure[112(104, 124) mmHg vs.109(98, 121) mmHg, 1 mmHg=0.133 kPa], triglyceride level [1.27(0.95, 1.81) mmol/L vs.1.09(0.79, 1.61) mmol/L], fas-ting blood glucose level [4.80(4.46, 5.01) mmol/L vs.4.48(4.16, 4.76) mmol/L] and fasting insulin level [21.27(16.21, 28.56) mmol/L vs.7.62(4.43, 10.83) mmol/L], and the differences were all statistically significant(all P<0.05). IR was a risk factor for NAFLD in obese children( OR=1.536, 95% CI: 1.049-2.247, P<0.05). Conclusions:Serious and abdominal obesity in children and adolescents is a major risk factor for the development of IR.HOMA-IR of obese children and adolescents is particularly serious during puberty.The obese children with IR are more likely to have metabolic disorders in blood glucose, serum lipid and blood pressure, and have the risk of NAFLD development.