Regulatory T cells inhibit FoxP3 to increase the population of tumor initiating cells in hepatocellular carcinoma.

PURPOSE: Tumor initiating cells (TICs) or cancer stem cells (CSCs) are considered to be the main culprit of hepatocellular carcinoma (HCC) initiation and progression, nevertheless the mechanism by which tumor microenvironment maintains the HCC 'stemness' is not fully understood. This study aims to investigate the effect of regulatory T cells (Tregs) on the TICs characteristics of HCC. METHODS: Immunocytochemistry, flow cytometry, real-time PCR, western blot, in vitro sphere-formation, and in vivo tumorigenesis assay were used to detect HCC 'stemness'. Additionally, after forced expression or inhibition of FoxP3, ß-catenin expression and HCC 'stemness' were investigated. RESULTS: Tregs enhanced the 'stemness' of HCC cells by upregulating TIC-related markers CD133, Oct3/4, Sox2, c-Myc, Klf4, Nanog, CD13, EpCAM, and inducting epithelial to mesenchymal transition (EMT), increasing TICs ratio, as well as promoting tumorigenic ability. Moreover, ß-catenin and c-Myc were upregulated in HCC cells after co-cultured with Tregs. HCC 'stemness' was inhibited after treatment with Wnt/ß-catenin pathway inhibitor. Furthermore, forced expression of FoxP3 resulted in increased GSK3ß, decreased ß-catenin and TIC ratio in HCC. In contrast, FoxP3 interference reduced GSK3ß, enhanced ß-catenin and TIC ratio of HCC. CONCLUSION: This study, for the first time, demonstrated that Tregs increased the population of TICs in HCC by inhibiting FoxP3 as well as promoting ß-catenin expression.

Vasodilating effect of suberoylanilide hydroxamic acid on isolated thoracic aorta of rats and its mechanisms / 中国药理学通报

Aim To investigate the vasodilating effect of suberoylanilide hydroxamic acid ( SAHA) on isolated thoracic aorta of rats and the possible mechanisms. Methods Isolated thoracic aorta of rats were used to perfuse, and to observe the effect of accumulated concentration of SAHA (0.3, 1, 3, 10 and 30 p,mol • L_1) on isolated thoracic aorta at basal state, KC1 and NE precontracted state. At the same time, L-NAME, Indo, PMA and SP were used to explore its possible mechanisms of relaxing isolated thoracic aorta, and to investigate the role of the Ca2 during the vasodilating process. Results SAHA could relax KC1 and NE precontracted isolated thoracic aorta of rats (P <0. 01), and the effect on endothelium-intact was stronger than that on endothelium-denuded thoracic aorta (P < 0.01), but there was no significant effect on thoracic aorta at basal state. The vasodilatory effect of SAHA could be inhibited by L-NAME, Indo and PMA (P < 0.05), while that could be promoted by SP ( P < 0. 01). SAHA could attenuate vasoconstriction induced by CaCl2 and NE through inhibiting extracellular Ca2 + influxe and intracellular Ca2 release in a concentration-dependent manner ( P < 0. 01). Conclusions The vasodilating mechanisms of SAHA may be related to the increased production of vasodilatory factors NO and PGt2, and the inhibition of PKC, and the inhibition of extracellular Ca2 + influxe and intracellular Ca2 + release.

Role of central PGE2 on sympathetic excitation in chronic heart failure / 中国病理生理杂志

AIM:To observe the effect of central prostaglandin E2(PGE2) on sympathetic activation in chronic heart failure (CHF) and to explore the underlying mechanism. METHODS:Male SD rats were subjected to coronary ar-tery ligation to induce heart failure (HF), and the intracerebroventricular infusion was performed by osmotic pump continu-ously. The rats in sham group and HF group were given artificial cerebrospinal fluid (0. 25 μL/h). The rats in HF plus treatment group was given celecoxib (CLB; 20 mg/h). After 4 weeks, the levels of PGE2 in cerebrospinal fluid ( CSF), the sympathetic nerve excitability and cardiac function were measured, and the changes of corticotropin-hormone releasing hormone ( CRH)-containing neurons activation and neurotransmitter contents in the hypothalamic paraventricular nucleus ( PVN) were also determined. RESULTS:Compared with the sham-operated rats, the HF rats had raised level of PGE2 in CSF, up-regulated renal sympathetic nerve activity and plasma norepinephrine, increased left ventricular end diastolic pres-sure, lung-to-body weight and right ventricular-to-body weight ratios, and decreased maximal increase and decreased rate of left ventricular pressure (P<0.05). In addition, the number of CRH positive neurons in PVN and the level of plasma ad-renocorticotropic hormone were higher in HF rats than those in sham-operated rats (P<0.05). After administration of CLB into the lateral ventricle of HF rats, the contents of PGE2 in CSF were significantly reduced, the number of activation CRH neurons in PVN was decreased, the excitability of sympathetic nerves was down-regulated and cardiac function was im-proved (P<0.05). Compared with the sham-operated rats, the content of glutamic acid in PVN of HF rats was increased, the content of γ-aminobutyric acid and the number of glutamate decarboxylase 67-positive neurons were decreased ( P<0.05). After the CLB was given, the above indexes were reversed (P<0.05). CONCLUSION:These findings indicate that in CHF, the increased central PGE2 may activate CRH-containing PVN neurons and contribute to the augmented sym-pathetic drive possibly by modulating the neurotransmitters within the PVN.

[Change rules of quantity and species of diatoms in Hunhe River in Shenyang].

OBJECTION: To observe the change rules of quantity and species of diatoms in Hunhe River in Shenyang and to provide technology and scientific evidence for drowning identification and the location of drowning in forensic investigation. METHODS: In 2011, different locations for collecting water samples were chosen in Hunhe River in Shenyang. Water samples were collected and variation of quantity and species of diatoms were observed every month. And variation of dominant species of diatoms was observed every week. RESULTS: The quantity, species and dominant species of diatoms in Hunhe River in Shenyang varied with different time and locations. The quantity and species of diatoms were lowest from December to February and gradually increased, reaching peak in May and second peak in October, and then gradually decreased. The dominant species of diatoms varied significantly adjacent two weeks at same location from April to November, but had little changes at different locations in same week from July to August. CONCLUSION: The change rules of quantity and species of diatoms are complicated and affected by various factors such as environment and hydrology. The change rules of species and quantity of diatoms should be considered in forensic investigation of drowning identification and the location of drowning.

Analysis of differentiation characters in osteosarcoma Mdr sublines / 中国骨伤

<p><b>OBJECTIVE</b>To explore the relationship between multidrug resistance (Mdr) and malignancy. To observe whether P-glycoprotein (P-gp) overexpression had the same bioactivity as osteogenic stem cell turning into more mature cell or more complex phenotype when parent cell line turned to Mdr phenotype.</p><p><b>METHODS</b>Six cell sublines of Mdr phenotype with different expression degree were analyzed. Stathmin generally identified in malignancy cell and stem cell, was a microtubule associated protein and the signal of differentiation in osteogenic stem cell. RT-PCR and hybridization in situ were used to analyze the relationship between Mdr1 mRNA and expression of Stathmin mRNA and VEGF mRNA.</p><p><b>RESULTS</b>Morphological and functional analysis of Mdr phenotype showed the P-gp-positive cell lines were more differentiated than the parent cells in terms of enhanced activity of cellular alkaline phosphatase. These subclones all displayed a decrease in potential malignancy such as tumor growth rate and metastatic ability. A significant negative correlation could be identified between Mdr1 mRNA and expression of VEGF and Stathmin mRNA.</p><p><b>CONCLUSION</b>Expression of Mdr1/P170 indicated osteosarcoma cells differentiated towards more mature state, which will develop the new research field of Mdr and supply the new research method of the function of P- gp.</p>

Study of gelatinized marrow stroma osteoblasts and true bone ceramic active bone / 中华创伤杂志(英文版)

<p><b>OBJECTIVE</b>To investigate a new method to construct tissue-engineering bone that will be applicable clinically.</p><p><b>METHODS</b>The cultured 5th generation rabbit bone marrow stroma osteoblasts (MSO) was dissolved in 3% sodium alginate solution (the final concentration of sodium alginate in the solution being 1%, and MSO, 5x10(6)/L), and then inoculated into prepared true bone ceramic (TBC) and gelatinized the bone by dribbling with calcium gluconate. The standard bone defect models were made in 48 adult New Zealand rabbit's both radius. Among the 48 rabbits, 24 were in Groups A and B, in which the left radius was implanted with gelatinized MSO-TBC (Group A) and right radius implanted with autograft-bone (Group B); and the other 24 were in control group whose left radius was implanted with non-gelatinized MSO-TBC (Group C) and right radius implanted with gelatinized TBC (Group D). Outcomes of the implanted bones were assessed by radiology, pathological histology, osteogenetic quantitative analysis, and biomechanics at 2, 4, 8, 12 weeks postoperatively.</p><p><b>RESULTS</b>In Groups A and B, a satisfactory bone reparation and bony union was noted within 12 weeks. In Groups C and D, bone reparation was not satisfied compared with Group A in terms of ostogenetic quantity and biomechanics.</p><p><b>CONCLUSIONS</b>Gelatinized MSO-TBC is an ideal artificial active bone that overcomes TBC shortcomings of fragileness and smooth surface that is not eligible for seed cell's adhesion. It is promising to put into clinical use extensively.</p>

Olfactory acuity and improvement of olfaction after total laryngectomy / 中华耳鼻咽喉头颈外科杂志

<p><b>OBJECTIVE</b>To test and evaluate the olfactory function of patient after total laryngectomy, seek to a practical method to ameliorate olfactory function and rise the qualitative character of these patients.</p><p><b>METHODS</b>Using the T&T olfactory examination to evaluate the olfactory function of 60 cases. Four cases olfactory mucosae were observed by electron microscope. Observing relation among the remains olfaction, the time after operation and whether or not undergone the voice reconstruction. And analyse the reasons of the above hyposomnia. Using the closing-mouth and nasal out-word airflow maneuver (CNOAM) as the intervention in the patients of tracheoesophageal fistula voice reconstruction after total laryngectomy to observe the amelioration after intervention.</p><p><b>RESULTS</b>It shows various degree of hyposmia and anosmia in the cases after total laryngectomy with or without tracheoesophageal fistula voice reconstruction, with significant deference (P < 0.01) compared to the normal person respectively. There are precisely correlation among the time after operation and whether or not undergone the voice reconstruction. The longer time leads to less remaining olfaction. The patients after total laryngectomy without tracheoesophageal fistula voice reconstruction have lost their olfaction thoroughly within 5 years. But for the patients after total laryngectomy with tracheoesophageal fistula voice reconstruction, they have a middle hyposmia within 5 years, with significant deference (P < 0.01) between the patients in 5 years and after 5 years. There were significant differences (P < 0.01) between the values of patients with and without tracheoesophageal fistula voice reconstruction. The ultrastructure of 4 cases of olfactory epithelium shows the apoptosis change. After the treatment of CNOAM, the remaining olfaction of most patients were improved, with significant deference (P < 0.01) compared to those before the treatment of CNOAM.</p><p><b>CONCLUSIONS</b>The proceed hypofunction of olfaction may be influenced by the reform of respiratory air, the extinction of air velocity bypass the nasal cavity and the apoptosis of epithelial cells in the patients after total laryngectomy. But if we give an early intervention study such as tracheoesophageal fistula voice reconstruction and CNOAM, the olfactory function may be maintenance. During the intervention, the ending of olfactory nerves may be get uninterrupt stimulation. This may help the patients keep a better existing quality than those fail to accept the interventions.</p>