Aim To investigate the changes of Na~+,K~+-ATPase activity and ? isoforms in rat neurons suffered from global ischemia.Methods The BCAL rat was induced by bilateral carotid artery ligation.The contents of H_2O,Na~+ and K~+ in brain tissue were detected.The method of spectrophotometry was applied to measure the activities of Na~+,K~+-ATPase.With immunohistochemical assay,the expression of the Na~+,K~+-ATPase ?_1,?_2,or ?_3 isoform was detected in neurons of rat hippocampus and cortex following ischemia.Results In the BCAL rats,the contents of H_2O and Na~+ were increased while the content of K~+ and the activity of Na~+,K~+-ATPase were decreased.The ?_1,?_2 or ?_3 isoform of Na~+,K~+-ATPase distributed in a tissue-specific fashion in neurons of hippocampus and cortex,in which ?_1 and ?_3 were abundant and ?_3 was more than ?_1,but ?_2 was much less.In BCAL rat,?_1 and ?_3 isoforms were significantly decreased in hippocampus and cortex neurons.Conclusion These results suggest that the changes of Na~+,K~+-ATPase ?_1 and ?_3 isoforms may be involved in the global cerebral ischemia

Objective To conduct a Meta-analysis to evaluate the efficacy and safety of interfer on-α for treating HBeAg-positive chronic hepatitis B in children.Methods PubMed and Chinese Biomedical Database were searched from the beginning to April 2006,and the references of eligible studies were manually screened.Randomized controlled trials published in the English and Chinese literature comparing interferon-α with non-antiviral interventions(placebo or no treatment)in children with chronic hepatitis B were eligible for inclusion.Studies were included if patients were treated for at least 3 months and followed up for at least 6 months after cessation of therapy.Two investigators independently assessed the quality and extracted the data.The methodological quality of trails was assessed by the Jadad-scale plus allocation concealment.Heterogeneity was examined by Chi-square test.Fixed effects model or random effects model was used to pool the data.Sensitivity analyses were used in the treatment course.Results Seven randomized controlled studies with a total of 360 child chronic hepatitis B virus carriers who were positive for hepatitis B surface antigen and hepatitis B e antigen werc identified.It was found by Meta-analysis that,compared with the control,at the end of therapy,interferon-α could significantly clear HBeAg[22.1%vs 6.7%,OR 3.56,95% CI(1.74, 7.28),P=0.000 5],HBV DNA[33.7% vs 12.6%,OR 3.50,95% CI(2.03,6.06),P＜0.01], HBsAg [6.5% vs 0.5%,OR 7.10,95% CI(1.52,33.12),P=0.01],and achieve HBeAg seroconversion [17.3% vs 2.9%,OR 5.62,95% CI(1.65,19.18),P=0.006],but was not more effective in HBsAg seroversion[2.0% vs 0,OR 3.55,95%CI(0.35,35.93),P=0.28]and alanine aminotransferase(ALT)normalization[24.2% vs 16.2%,OR 1.72,95% CI(0.84,3.52), P=0.14].Conclusions Interferon a may be efficacious in clearance of HBeAg,HBV DNA and HhsAg, and achievement of HBeAg seroversion.Little evidence is available on HBsAg seroversion and ALT normali zation.Rigorously designed large sample size randomized double blind clinical trials with large sample size are required to further confirm and support the conclusion.

Aim To observe effects of hypertension on contractile/diastolic function and calcium transient in rats ventricular myocytes. Methods The model of one-kidney-one-clip (1k1c) hypertensive rat was pre-pared by partially ligating the left renal artery and removing the right kidney. Left ventricular myocytes were enzymatically isolated. Then the contraction and calcium transient of a single cell from both normal and renovascular hypertensive rats were observed by a video-based motion edge-detection system simultaneously. Effects of calcium in various concentrations on contractile/diastolic function and calcium transient in ventricular myocytes from renovascular hypertensive rats were assessed in the same way. Results Compared with normal cardiac myocytes, the shorting amplitude and the contractile and diastolic velocity were increased significantly in 1k1c hypertensive rat cardiac myocytes. However their intracellular calcium in contractile and diastolic periods, the extent of calcium transient and the parameters of intracellular calcium dynamics were unchanged. But the extracellular calcium of different concentrations could shift the Fura-2 fluorescence ratio-cell shorting amplitude curve from hypertension rat myocytes to the left compared with that from normal rats. Conclusions The hypertension increases the contractility of rat cardiac myocytes, which is due to raising their sensitivity to calcium.

Objective To investigate the expression and clinical significance of tumor-associated carbohydrate anti-gen sTn in endometrium from patients with atypical hyperplasia of endometrium, endometrial carcinoma and benign the uter-ine lesion. And to analyze the association between sTn and the pathologic stage and grade of endometrial carcinoma. Meth-ods sTn was detected by immunohistochemistry in 193 patient samples, including 93 subjects with endometrial carcinoma, 50 subjects with atypical hyperplasia of endometrium and 50 subjects with benign uterine lesions. Results The positive sTn rate in endometrial carcinoma group was 73.1%,which was significantly different from that in atypical hyperplasia endo-metrium group (32%) and that in benign uterine lesion group (14%), (P＜0.017). sTn positive rate in patients with endometri-al carcinoma was 78.8%which is higher than that in patients with other diseases (P＜0.008 ). The positive rate of sTn in pa-tients with poorly differentiated endometrial carcinoma was significantly higher than that in patients with highly differentiat-ed endometrial carcinoma (P＜0.017). The positive rate of sTn in tissues of tumor infiltration with depth of more than half of myometrial was significantly higher than in tissues of tumor infiltration with depth of equal or less than half of myometrial (P＜0.05). But there was no significant difference in sTn positive rate between the endometrial carcinoma groups with and without lymph node metastasis (P>0.05). Conclusion sTn may play a role in the tumorigenesis and development of endo-metrial carcinoma.

The study aims to identify the role of cAMP-PKA pathway in the group Ⅱ metabotropic glutamate receptors (mGluRs)-mediated regulation of respiratory rhythm from the brainstem slice. Neonatal (aged 0-3 d) Sprague-Dawley rats of either sex were used. The brainstem slice containing the medial region of the nucleus retrofacialis (mNRF) and the hypoglossal nerve rootlets was prepared, and the surgical procedure was performed in the modified Kreb's solution (MKS) with continuous carbogen (95% O2 and 5% CO2) bubbling, and ended in 3 min. Respiratory rhythmical discharge activity (RRDA) of the hypoglossal nerve rootlets was recorded by suction electrode. Eighteen brainstem slice preparations were divided into 3 groups. In group 1, group Ⅱ mGluRs specific antagonist (2S)-α-ethylglutamic acid (EGLU) was added into the perfusion solution for 10 min. In group 2, after application of Forskolin for 10 min, washout with MKS, the slice was perfused with Rp-cyclic 3', 5'-hydrogen phosphorothioate adenosine triethylammonium salt (Rp-cAMPS) alone for another 10 min. In group 3, after application of Rp-cAMPS for 10 min, additional EGLU was added into the perfusion for another 10 min. The results showed EGLU shortened respiratory cycle (RC), but the changes of integral amplitude (IA) and inspiratory time (TI) were not statistically significant. Forskolin induced significant decreases in RC, and increased TI, IA. Rp-cAMPS could make the opposite effect compared with the changes of RRDA with Forskolin. The effect of EGLU on the RRDA was inhibited after blocking the cAMP-PKA pathway. Taken together, cAMP-PKA pathway may play an important role in the group Ⅱ mGluRs-mediated regulation of RRDA in the brainstem slice of neonatal rats.
Animals ; Animals, Newborn ; Brain Stem ; physiology ; Cyclic AMP-Dependent Protein Kinases ; metabolism ; Female ; In Vitro Techniques ; Male ; Rats ; Rats, Sprague-Dawley ; Receptors, Metabotropic Glutamate ; physiology ; Respiration ; Signal Transduction ; physiology

BACKGROUNDHypoxia-inducible factor-1alpha (HIF-1alpha) is one of the pivotal mediators in the response of lungs to decreased oxygen availability, and increasingly has been implicated in the pathogenesis of pulmonary hypertension. Vascular endothelial growth factor (VEGF), a downstream target gene of HIF-1alpha, plays an important role in the pathogenesis of hypoxic pulmonary hypertension and hypoxic pulmonary artery remodelling. In this study, we investigated the dynamic expression of HIF-1alpha and VEGF in pulmonary artery of rats with hypoxia-induced pulmonary hypertension.

METHODSForty male Wistar rats were exposed to hypoxia for 0, 3, 7, 14 or 21 days. Mean pulmonary arterial pressure (mPAP), vessel morphometry and right ventricle hypertrophy index (RVHI) were estimated. Lungs were inflated and fixed for in situ hybridisation and immunohistochemistry.

RESULTSmPAP values were significantly higher than the control values after 7days of hypoxia [(18.4 +/- 0.4) mmHg, P < 0.05]. RVHI developed significantly after 14 days of hypoxia. Expression of HIF-1alpha protein increased in pulmonary arterial tunica intima of all hypoxic rats. In pulmonary arterial tunica media, HIF-1alpha protein was markedly increased by day 3 (0.20 +/- 0.02, P < 0.05), reached the peak by day 7, then declined after day 14 of hypoxia. HIF-1alpha mRNA increased significantly after day 14 of hypoxia (0.20 +/- 0.02, P < 0.05). VEGF protein began to increase markedly after day 7 of hypoxia, reaching its peak around day 14 of hypoxia (0.15 +/- 0.02, P < 0.05). VEGF mRNA began to increase after day 7 of hypoxia, then remained more or less stable from day 7 onwards. VEGF mRNA is located mainly in tunica intima and tunica media, whereas VEGF protein is located predominantly in tunica intima. Linear analysis showed that HIF-1alpha mRNA, VEGF and mPAP were correlated with hypoxic pulmonary artery remodelling. HIF-1alpha mRNA was positively correlated with VEGF mRNA and protein (P < 0.01).

CONCLUSIONHIF-1alpha and VEGF are both involved in the pathogenesis of hypoxia-induced pulmonary hypertension in rats.

Animals ; Blood Pressure ; Chronic Disease ; Hypertension, Pulmonary ; etiology ; Hypertrophy, Right Ventricular ; etiology ; Hypoxia ; complications ; metabolism ; Hypoxia-Inducible Factor 1, alpha Subunit ; Male ; Pulmonary Artery ; metabolism ; RNA, Messenger ; analysis ; Rats ; Rats, Wistar ; Transcription Factors ; genetics ; physiology ; Vascular Endothelial Growth Factor A ; genetics ; physiology

OBJECTIVETo investigate the effects of advanced glycosylation end products (AGEs) modified bovine serum albumin (AGE-BSA) on the expression of reactive oxygen species (ROS) and monocyte chemoattractant protein-1 (MCP-1) in human renal mesangial cells (HRMCs).

METHODSHRMCs were cultured in vitro with medium containing different doses of AGE-BSA or BSA (50,100, 200, 400 mg/L) for 48 hours, or with AGE-BSA (200 mg/L) for different times (12, 24, 48, 72 h). Immunocytochemistry assay was used to estimate the protein level of RAGE. The ROS in cells were measured by flow cytometry and the mRNA expression of MCP-1 were analyzed by semi-quantiative reverse transcription-polymerase chain reaction (RT-PCR) after treatment with AGE-BSA or BSA.

RESULTSThe protein level of RAGE was upregulated in the HRMCs with AGE-BSA. The expression of ROS and MCP-1 significantly enhanced by incubation of AGE-BSA in a time- and dose-dependent manner. The effects of AGE-BSA-induced up-regulation of ROS and MCP-1 level was significantly blocked by neutralizing antibodies to RAGE, while the expression of ROS and MCP-1 stood nearly unchanged after cultured with huamn IgG.

CONCLUSIONThe expression of ROS and MCP-1 in HRMCs is induced by AGE-BSA through RAGE, which may have potential effects in the pathgenic mechanism of diabetic nephropathy.

Cells, Cultured ; Chemokine CCL2 ; metabolism ; Glycation End Products, Advanced ; pharmacology ; Humans ; Mesangial Cells ; drug effects ; metabolism ; Oxidative Stress ; drug effects ; Reactive Oxygen Species ; metabolism ; Receptor for Advanced Glycation End Products ; Receptors, Immunologic ; metabolism ; Serum Albumin, Bovine ; pharmacology

Objective To investigate the efficacy of AI-SONICTM Thyroid system, a version 2.0 artificial intelligence (AI) automatic detection system, in the preoperative ultrasound diagnosis of thyroid nodules, and to evaluate the application value of AI automatic detection system version 2.0 in the differential diagnosis of benign and malignant thyroid nodules by comparing with the subjective diagnosis conclusions of sonographers with different seniorities. Methods A total of 247 patients (325 thyroid nodules) admitted to the Department of General Surgery in our hospital from Aug. 2019 to Jan. 2020 were selected for this study. All patients underwent routine ultrasound examinations by a senior sonographer with 13 years of experience in thyroid ultrasound diagnosis and a junior sonographer with 4 years of work experience. At the same time, the patients were also examined by another sonographer with 20 years of work experience using AI automatic detection system version 2.0, without knowing the diagnosis conclusions of the above two sonographers. Kappa test was used to evaluate the consistency of the results of routine ultrasound examination of sonographers with different seniorities and AI automatic detection system version 2.0 and the postoperative pathological results. Results The postoperative pathology confirmed 229 malignant nodules and 96 benign nodules. The sensitivity, specificity and accuracy in the diagnosis of benign and malignant thyroid nodules were 85.15% (195/229), 66.67% (64/96) and 79.69% (259/325), 93.45% (214/229), 79.17% (76/96) and 89.23% (290/325), and 92.58% (212/229), 71.88% (69/96) and 86.46% (281/325) for junior sonographer, senior sonographer and AI automatic detection system version 2.0, respectively. The Kappa consistency test results showed that the diagnosis result of senior sonographer was highly consistent with the pathological diagnosis result (Kappa value 0.78, P＜0.01), while the diagnosis results of junior sonographer and AI automatic detection system version 2.0 were generally consistent with the pathological diagnosis result (Kappa values 0.55 and 0.74, both P＜0.01). Conclusion The sensitivity, accuracy and specificity of the AI automatic detection system version 2.0 AI-SONICTM Thyroid in diagnosing benign and malignant thyroid nodules are similar to those of routine ultrasound examination by senior sonographers, and the system might be a reliable auxiliary means for preoperative evaluation of benign and malignant thyroid nodules.

Objective: To explore the application value of artificial intelligence (AI) automatic detection system in preoperative ultrasonic diagnosis of thyroid nodules. Methods: Totally 98 patients with 137 thyroid nodules admitted to the General Surgery Department of Changzheng Hospital of Naval Medical University (Second Military Medical University) from April 2019 to July 2019 were enrolled in this study. Pathological data and ultrasonic diagnosis results were retrospectively analyzed. All patients underwent conventional ultrasonography and AI automatic detection before surgery. The diagnoses for benign and malignant thyroid nodules were compared between conventional ultrasonography and AI automatic detection system, which were based on the postoperative pathology. The sensitivity, specificity and accuracy of the two examination methods were calculated, and Kappa coefficient was performed to measure the consistency between the two methods and postoperative pathological diagnosis. Results: The sensitivity, specificity and accuracy of conventional ultrasonography in diagnosis of benign and malignant thyroid nodules were respectively 93.75% (90/96), 80.49% (33/41) and 89.78% (123/137), and those of AI automatic detection were 89.58% (86/96), 68.29% (28/41) and 83.21% (114/137). There was substantial coefficience between conventional ultrasonography and pathological diagnosis results (Kappa=0.75, P<0.001), and that was fair coefficience between AI automatic detection system and pathological diagnosis results (Kappa=0.59, P<0.001). Conclusion: The sensitivity and accuracy of AI automatic detection system are slightly lower than but close to those of conventional ultrasonography in differentiating benign from malignant thyroid nodules. AI automatic detection system can be used as an effective supplement to assist conventional ultrasonography for preoperative assessment of thyroid nodules.

Objective To study the difference of gene expression profile between the radioresistant human nasopharyngeal carcinoma cell line CNE-2R and CNE-2,and to screen the signaling pathway associated with radioresistance of nasopharyngeal carcinoma.Methods The radioresistant nasopharyngeal carcinoma cell line CNE-2R was constructed from the original cell line CNE-2.CNE-2R and CNE-2 cells were cultured and administered with 60Co γ-ray irradiation at the dose of 400 cGy for 15 times.Human-6v 3.0 whole genome expression profile was used to screen the differentially expressed genes.Bioinformatic analysis was used to identify the pathways related to radioresistance.Results The number of the differentially expressed genes that were found in these 2 experiments was 374.The Kegg pathway and Biocarta pathway analysis of the differentially expressed genes showed the biological importance of Toll-like receptor signaling pathway and IL-1 R-mediated signal transduction pathway to the radioresistance of the CNE-2R cells and the significant differences of 13 genes in these 2 pathways,including JUN,MYD88,CCL5,CXCL10,STAT1,LY96,FOS,CCL3,IL-6,IL-8,IL-1α,IL-1B,and IRAK2(t=13.47-66.57,P<0.05).Conclusions Toll-like receptor signaling pathway and IL-1R-mediated signal transduction pathway might be related to the occurrence of radioresistance.