BACKGROUND:Bone marrow mesenchymal stem cells can differentiate into osteoblasts under inducing condition that Zouguiwan and Youguiwan coordinate inducers, but the mechanism remains to be discussed. OBJECTIVE:To observe the effects of serum containing Zuoguiwan and Youguiwan on transforming growth factorβ1 and its signal transduction protein Smad2/3 message expression during the osteogenic differentiation of bone marrow mesenchymal stem cells. METHODS:A whole bone marrow adherence method was adopted to isolate and cultivate bone marrow mesenchymal stem cells from rats. The cellcultivation was processed in five groups:bone marrow mesenchymal stem cells were respectively cultured with blank serum, serum containing Zouguiwan, serum containing Youguiwan, positive serum containing progynova+inducer (dexamethasone, vitamin C, andβ-glycerophosphate), and inducer. Western blot was applied to detect the expression of type I col agen. The immunohistochemical assay was utilized to test transforming growth factorβ1 and Smad2/3 expression in the osteoblasts. RESULTS AND CONCLUSION:It was apparently more significant for serum containing Zuoguiwan and Youguiwan on type I col agen, transforming growth factorβ1 and Smad2/3 expression, compared with blank serum group and inducer group (P<0.05);moreover, serum containing Zuoguiwan was better than serum containing Youguiwan (P<0.05). Both of serum containing Zuoguiwan and Youguiwan are able to promote osteogenic differentiation of bone marrow mesenchymal stem cells. Moreover, Zuoguiwan is much more effective indicating that this method of traditional Chinese medicine about nourishing kidneys can be better to promote osteogenic induction of bone marrow mesenchymal stem cells.

Objective To study the effects of prenatal cocaine exposure on the development of offspring's brains by building a murine model. Methods We weighted the body weight and brain weight of offspring on P10 from COC and SAL groups and observed the development of neuron and astrocyte in cerebral cortex by toluidine blue staining and immunohistochemistry. Results The brain weight and body weight from COC were both reduced on P10 compared with SAL group.We discovered prenatal cocaine exposure induced polarity disorder and dysplasia of neuron in cerebral cortex;the number of the astrocytes in corpus callosum and hippocampus regions decreased.Conclusion\ Pregnatal cocaine exposure can result in abnormal development of cerebral cortex of offsprings which may play an important role on cocaine induced abnormal behavior.\;[

AIM To develop a murine model for investigating the effects of prenatal cocaine exposure on the development of fetal nerve system. METHODS A nutritionally paired control group of dams injected with saline and pair fed with the COC dams were set up. Another two groups were COC groups injected with cocaine HCl and SAL group administrated with saline. After injection twice daily during gestation days 8～17,mice were decapitated on E17 and blood and brain samples were collected for pharmacological analysis and neurotransmitter analysis by HPLC.RESULTS Pharmacological analysis revealed that cocaine was found in maternal and fetal plasma at 15 min following ip administration to embryonic day E17 pregnant mice. Though COC dams and SPF dams had the same feeding condition, compared with the latter, the former had higher maternal concentrations of DA and 5 HT, lower fetal weight, brain weight, striatum weight and higher concentrations of DA and 5 HT in striatum, P

AIM To investigate the effects of in utero cocaine exposure on the fetal development, when fetuses were exposed to equal total dose but different dose and timing. METHODS Pregnant dams were randomly separated into three groups: SAL, COC20 and COC40. On E17, recorded body weight, brain weight and striatum weight of all groups, and examined the concentrations of DA and 5 HT in fetal striatum by HPLC. RESULTS Body weight of SAL, COC40, COC20 groups decreased progressively in turns. Brain weight of COC20 group and COC40 group was lower than that of SAL. Only the brain/body ratio of COC40 was decreased ( P

Objective To investigate the molecular mechanism of osteoclast differentiation induced by staphylococcal lipoteichoic acid (LTA-sa). Methods Raw264.7 cells were treated with LTA-sa in a concentration of 200 ng/mL for 0, 5, 10, 20, 40, 60 min and 0, 1, 2, 3 days respectively, and the proteins in signaling pathways associated with osteoclast differentiation were measured with western blot. In addition, Raw264.7 cells were treated with different concentrations of LTA-sa (100, 200 and 400 ng/mL) and PBS for 0, 1, 2, 3 days, the expression of TNF-α, IL-1α and IL-6 was detected with Enzyme linked immunosorbent assay (ELISA). Results (1)Western blot showed that, under stimulation of LTA-sa, IκB-α decreased at 5 min and 10 min, while the phosphorylation of nuclear factor κB increased at 10 min . In addition , NFATc1 increased in 2 and 3 days gradually. The above results were statistically analyzed, and the difference was significant in statistics (P < 0.001). (2)ELISA showed that the expression of IL-6 increased in 2 and 3 days along with the increasing concentration and prolonging stimulation time of LTA-sa. Data were statistically analyzed, the difference was significant in statistics (P < 0.001). Conclusion LTA-sa promotes osteoclast differentiation through the NF-κB signaling pathway and the secretion of IL-6.

Background and purpose:Researches had indicated that about over 85%of malignant tumors highly express telomerase activity. So telomerase has become one of the important methods in the research field of tumor diagnosis and treatment. Nowadays, several reports about malignant tumor which over expresses hTERT targeting imaging with radionuclide labeled hTERT ASON had been published. In these reports, high quality of pictures can hardly be acquired because of poor anatomical and spacial resolution in nuclear imaging itself. Accordingly, in this study, we developed a method of detecting human telomerase in vivo with magnetic resonance imaging (MRI) and evaluate its feasibility. Methods:Firstly, Uniformly phosphorothioate-modified human telomerase reverse transcriptase antisense oligonucleotide (hTERT ASON) was labeled with Gd3+ through the bifunctional chelator 1, 4, 7, 10-tetraazacyclododecane-N, N’, N’’, N’’’-tetraacetic acid (DOTA) and iv vitro experiments were performed to characterize the antisense probes (for biodistribution and cellular uptake, 99mTc-DOTA-ASON was used in stead of Gd-DOTA-ASON). Then Gd-DOTA-ASON was injected intraperitoneally in pulmonary adenocarcinoma A375 nude mice tumor-bearing BALB/c for in vivo imaging using 7.0 T Micro MRI periodically, tumors and their surrounding tissues were defined as region of interest (ROI) to calculate the signal to noise ratio (SNR) of tumor to muscle using Gd-DTPA as control. Finally, immunohistochemical analysis of telomerase activity of each xenograft was operated 2 days after imaging. Results:The binding efficiency of Gd-DOTA-ASON reached was as high as 65%(63.2±2.4, n=6). And it can maintain 61%in fresh human serum and normal saline at 37℃over 24 h;A375 cells showed an uptake of 8.5%when incubated with 99mTc-DOTA-ASON;In comparing with DOTA-ASON and Gd-DTPA, cells transected with Gd-DOTA-ASON had higher SI when performed MRI with T1WI. The hTERT-expressing xenografts were obviously enhanced by Gd-DOTA-ASON at 0.5-6 h after injection and the SNR can reach 2.37, whereas obvious enhancement only could be found within 2 h after injection of Gd-DTPA. Both labeled and non-labeled antisense probes can suppress the activity of telomerase of A375 cells either in vitro or in vitro. Conclusion:Our research offers proof that Gd-DOTA-ASON can be used as tumor specific targeting MR probe for diagnosing malignant tumors with high expression of telomerase.

OBJECTIVE: To observe the expression of aquaporin 4 (AQP4) in diffuse brain injury (DBI) of rats and to explore the corresponding effect of AQP4 for brain edema. METHODS: The rat model of DBI was established using Marmarou's impact-compression trauma model. Brain water content was measured by dry-wet weight method. Blood-brain barrier permeability was evaluated by Evans blue (EB) staining. Immunohistochemical method was used to observe the expression of AQP4. RESULTS: Brain water content increased after 3 h and peaked at 24 h after DBI. Brain EB content significantly increased and peaked at 12 h after DBI. The expression of AQP4 significantly increased after 3 h and peaked at 24 h after DBI, and the number of AQP4 positive astrocytes increased. CONCLUSION The increment of the permeability of blood-brain barrier and the expression of AQP4 may contribute to the development of brain edema in rat DBI. The change of AQP4 expression in astrocytes may also contribute to determine DBI.
Animals ; Aquaporin 4/metabolism* ; Astrocytes ; Blood-Brain Barrier/metabolism* ; Brain ; Brain Edema/metabolism* ; Brain Injuries/metabolism* ; Cell Membrane Permeability/genetics* ; Disease Models, Animal ; Permeability ; Rats ; Water

Objective:To investigate the clinical characteristics and the risk of major adverse cardiac events within 1 year of middle-aged and elderly in-patients with acute decompensated and mid-range ejection fraction heart failure(HF)in the medical alliance setting.Methods:A retrospective cohort study was conducted among a total of 180 in-patients with acute decompensated heart failure in Cardiovascular Hexi Hospital Consulting Area of Tianjin Chest Hospital.According to ejection fraction measured by echocardiogram, the in-patients were classified into three groups: heart failure with reduced ejection fraction(HFrEF)group(n=70, 38.9%), HFmEF group(n=50, 27.8%), and heart failure with preserved ejection fraction(HFpEF)group(n=60, 33.3%). Clinical feature and 1-year prognosis between different groups were compared.Results:Univariate Cox regression analysis of 1-year all-cause death and cardiovascular death showed that there was no significant difference between HFrEF group and HFmEF group, HFpEF group and HFmEF group(all P>0.05); 1-year readmission analysis of heart failure showed that 47.1%(33 cases)of HFrEF group was higher than 24.0%(12 cases)of HFmEF group, 48.3%(29 cases)of HFpEF group was higher than HFmEF group( HR=2.307, 2.368, 95% CI: 0.187-4.480, 1.207-4.644, respectively, all P<0.05); The major 1-year cardiovascular events were 57.1%(40 cases)higher in the HFrEF group than 34.0%(17 cases)in the HFmEF group( HR=2.053, 95% CI: 0.187-4.408, P< 0.05). Multivariate analysis showed that the 1-year risk of major cardiovascular events was significantly different between HFmEF group and HFpEF group( HR=0.477, 95% CI: 0.241-0.941, P< 0.05). Pulmonary heart disease( P< 0.05), atrial flutter and/or atrial fibrillation( P< 0.01), New York Cardiology class Ⅳ( P< 0.01)were risk factors for death.Hypertension and cor pulmonale were the risk factors for readmission in patients with heart failure(all P< 0.01). Conclusions:The clinical characteristics of inpatients with HFmEF in the medical alliance setting tended to be consistent with those with HFrEF, while the feature of ischemic heart disease was more prominent in HFmEF.The 1-year risk of heart failure readmission in HFmEF group was significantly lower than that in HFpEF and HFrEF group, and the risk of all-cause mortality and cardiovascular mortality at 1 year was not significantly different among the three groups.

The aim of the experiments was to develop and characterize a murine model for investigating the effects of prenatal cocaine exposure on the mother and fetus. Pregnant mice were separated into three groups: group 1 was treated with cocaine HCl at 10 mg/kg twice daily (COC); group 2 was treated with saline at 10 ml/kg twice daily (SAL); and group 3 was pair-fed with the COC dams and was injected with saline following the same schedule (SPF) from embryonic day (E) 8 to 17. We utilized high-pressure liquid chromatography (HPLC) with UV detector and electrochemical detector to test the concentrations of cocaine, dopamine and serotonin, as well as HE staining to observe morphological alterations of liver and placenta. Though less food intake and lower weight gain were observed in COC and SPF groups but not in SAL dams, lower fetal body weight and brain weight were only seen in COC offspring. Pharmacological analysis revealed that cocaine was found in fetal plasma at 15 min following intraperitoneal administration on E17, accompanied with elevated concentrations of dopamine (DA) and serotonin (5-HT) in fetal brain. We also observed morphological changes in liver and placenta of cocaine-exposed fetuses. The present study indicates that pregnancy cocaine exposure can lead to maternal undernutrition and developmental abnormality of the fetal brain, liver and placenta. It is suggested that the developmental abnormality of the fetuses induced by cocaine is due to the toxicological effect of cocaine but not to maternal undernutrition.
Animals ; Brain ; metabolism ; pathology ; Cocaine ; adverse effects ; blood ; Disease Models, Animal ; Dopamine ; metabolism ; Female ; Fetus ; drug effects ; pathology ; Liver ; pathology ; Malnutrition ; Maternal Exposure ; adverse effects ; Maternal Nutritional Physiological Phenomena ; Mice ; Mothers ; Placenta ; pathology ; Pregnancy ; Serotonin ; metabolism

Objective To summarize the experience in open surgery for huge adrenal tumors in order to improve its safety and efficiency of this complicated surgical procedure. Methods Fortyfour consecutive patients with huge adrenal tumors underwent open surgery with mean long tumor diameter of 13 cm (9-34 cm), and autologous blood transfusion was prepared in routine. It was analyzed retrospectively for clinical data, perioperative complications and the effective and safety results of this procedure. Results The incision was oblique in lumbar region in 5 cases, subcostal unilaterally in 32 cases and abdomino-thoracic joint in 7 cases. There were 27 malignant tumors (61.4%) in 44 cases, 3 with hepatic invasion, 6 with thrombi extending into inferior vena cava, among which 2 needed translocation of artificial blood vessels and 3 needed cardio-pulmonary bypass. The mean blood loss was 1309 ml (100-3000 ml) in 41 cases(93.2%)and the autologous blood transfusion was used in 20 case (45.5%). There were 1 diaphragmatic injury, 1 pleura injury, 3 hemorrhage in large amount more than 15 000 ml and 2 peritoneal cavity infection.There were no perioperative deaths and 42 tumors (95.5%) were curatively resected. Conclusions Open surgery for huge adrenal tumors is a complicated surgical technique with high risk and large amount of blood loss. The key points to success are proper selection of incision, routine autologous blood transfusion, perfect surgical skills and good cooperation between different specialties.