Total nitrogen and L-hydroxyproline content of adipose tissue from various sites in rats of different ages

The total nitrogen and L-hydroxyproline concentrations of two intra-abdominal and two subcutaneous sites of adipose tissue were measured in male and female Wistar rats. At 100 g body weight the nitrogen concentration of intra-abdominal adipose tissue was slightly lower than that of subcutaneous adipose tissue. As age advanced beyond sexual maturity, the nitrogen content of intra-abdominal adipose tissue was steadily reduced, whereas that of subcutaneous tissue did not change. The L-hydroxyproline concentration of intra-abdominal adipose tissue was also lower than in subcutaneous adipose tissue. In male rats it rose in most adipose tissue sites when sexual maturity was reached, then fell with advancing age. In female rats the pattern of change was less consistent, but the l-hydroxyproline content of intra-abdominal adipose tissue was reduced as age increased.     

Glutathione and GSH-dependent enzymes in patients with liver cirrhosis and hepatocellular carcinoma

We investigated glutathione level, activities of selenium independent GSH peroxidase, selenium dependent GSH peroxidase, GSH S-transferase, GSH reductase and the rate of lipid peroxidation expressed as the level of malondialdehyde in liver tissues obtained from patients diagnosed with cirrhosis or hepatocellular carcinoma. GSH level was found to be lower in malignant tissues compared to adjacent normal tissues and it was higher in cancer than in cirrhotic tissue. Non-Se-GSH-Px activity was lower in cancer tissue compared with adjacent normal liver or cirrhotic tissue, while Se-GSH-Px activity in cancer was found to be similar to its activity in cirrhotic tissue and lower compared to control tissue. An increase in GST activity was observed in cirrhotic tissue compared with cancer tissue, whereas the GST activity in cancer was lower than in adjacent normal tissue. The activity of GSH-R was similar in cirrhotic and cancer tissues, but higher in cancer tissue compared to control liver tissue. An increased level of MDA was found in cancer tissue in comparison with control tissue, besides its level was higher in cancer tissue than in cirrhotic tissue. Our results show that the antioxidant system of cirrhosis and hepatocellular carcinoma is severely impaired. This is associated with changes of glutathione level and activities of GSH-dependent enzymes in liver tissue. GSH and enzymes cooperating with it are important factors in the process of liver diseases development.     

基于低场核磁共振T1-T2谱技术定性和定量分析罗非鱼各组织的方法探究

形体指标的检测是鱼类常规营养评定的重要组成部分。目前,分析鱼类组织体重比的方法会对鱼体造成损伤,且取样过程复杂,迫切需要无损快速检测技术分析鱼体组织占比。低场核磁共振技术具有快速无损检测的特点,已有研究利用低场核磁共振T1谱技术检测小鼠内脂肪组织体积,但尚未有研究应用低场核磁共振T1-T2谱技术对鱼类各组织器官进行定性和定量分析。基于此,利用低场核磁共振T1-T2谱技术,将分离后的罗非鱼的肌肉组织、腹腔脂肪组织、肝脏组织、肠道组织单独以及混合后进行扫描分析,结果发现利用低场核磁共振T1-T2谱技术可以分离罗非鱼肌肉组织和腹腔脂肪组织,但是无法区分肝脏组织和肠道组织。进一步对能够实现分离的肌肉组织和脂肪组织建立定量分析的模型,分析罗非鱼组织信号强度与组织重量相关性,结果显示,肌肉组织相关性R2=0.974 3,腹腔脂肪组织相关性R2=0965 0。并利用罗非鱼活体验证了肌肉组织定量分析模型的可靠性,将活体扫描肌肉信号大小转换成肌肉组织重量并分析其与全鱼体重相关性,结果显示肌肉组织重量与体重相关性R2=0.806 9。研究表明,在鱼类营养代谢研究中,可以利用低场核磁共振T1-T2谱技术快速无损地定量分析鱼体内肌肉组织含量。     

Convertible adipose tissue in mice

Summary Ability to express uncoupling protein (UCP) and establish UCP-dependent thermogenesis was analyzed in anatomical areas of mice that are generally considered to be white adipose tissue: mesenterial, perimetral, epididymal, inguinal, and superficial layer of interscapular white adipose tissue. The mice were acclimatized for 1 week to 4° C; the following week they were exposed to cold stress (1 h at-20° C, 2–3 times daily). In such conditions in inguinal adipose tissue, slot-blot analysis detected significant amount of UCP mRNA and lipoprotein lipase mRNA. Immuno-electron-microscopic localization of UCP showed that developed mitochondria of cold-stressed inguinal adipocytes contained UCP in the same amount as uncoupled (UC)-mitochondria of brown adipocytes. Morphological and morphometrical analysis showed that such inguinal adipose tissue appeared as brown adipose tissue. Since in control mice, inguinal adipose tissue was UCP-negative and tissue appeared as white adipose tissue, the duration of this white-to-brown adipose tissue conversion was analyzed. Mice, cold stressed for 1 week, were rewarmed at 28° C and their inguinal adipose tissue was analyzed in comparison with interscapular brown adipose tissue and epididymal white adipose tissue for another 37 days. During that time inguinal adipocytes ceased expressing UCP mRNA; UC-mitochondria in inguinal adipocytes were destroyed and replaced with common, C-mitochondria; and UCP was undetectable immunohistochemically. Adipocytes accumulated lipids, and the tissue morphologically once again resembled white adipose tissue. Described changes showed that besides typical brown and white adipose tissue in mice, there existed a third type of adipose tissue described as convertible adipose tissue.     

Determination of tissue plasminogen activator by an enzyme-immunoassay method

The sensitive assay method of tissue plasminogen activator was established by an enzyme-immunoassay method, and discriminates tissue (nonurokinase) type plasminogen activator from urokinase. The sensitivity was 0.1 ng/assay tube, and the plasma concentration of tissue plasminogen activator in normal healthy subjects was 1.22 +/- 0.25 ng/ml. Distribution of tissue plasminogen activator was examined in normal tissue. A melanoma cell line was employed as cell culture medium for determination of tissue plasminogen activator.     

Gender differences in tumor necrosis factor alpha and leptin secretion from subcutaneous and visceral fat tissue

Tumor necrosis factor alpha (TNFalpha) and leptin concentrations were determined in the abdominal subcutaneous and visceral (omental) adipose tissue of patients undergoing elective open-abdominal surgery and compared with their body mass index. The concentration of leptin did not differ significantly between women and men, being high in subcutaneous fat tissue and low in visceral fat tissue. TNFalpha concentration in subcutaneous fat tissue was approximately the same in both genders, but it was significantly lower in visceral fat tissue of women and unchanged in visceral fat tissue of men. A significant correlation between BMI and leptin was found in the two fat tissue compartments of both genders, but the correlation between BMI and TNFalpha was found only in subcutaneous fat tissue of women.     

High-energy diets produce different effects on fatty acid synthesis in brown adipose tissue, white adipose tissue and liver in the rat

The influence of feeding rats a high-energy diet for 7 days on fatty acid synthesis in brown adipose tissue, white adipose tissue and liver of the rat was investigated. The incorporation of 3H2O and [U-14C]glucose into fatty acid was measured in vivo. The rats fed the high-energy diets had higher rates of fatty acid synthesis in white adipose tissue than the controls fed on chow, while fatty acid synthesis in brown adipose tissue and liver was either decreased or unchanged relative to that of controls fed on chow. After an oral load of [U-14C]glucose the incorporation of radioactivity into tissue fatty acid was several-fold higher in brown adipose tissue than in white adipose tissue in rats fed on chow. In rats fed the high-energy diets, incorporation of radioactivity into fatty acid in brown adipose tissue was decreased while that into white adipose tissue was either increased (Wistar rats) or unchanged (Lister rats).     

Endogenous steroid production in cellular and subcellular fractions of rat testis after prolonged treatment with gonadotropins.

Steroid production and enzyme activities were examined in preparations of whole testis tissue, isolated interstitial tissue and seminiferous tubules obtained from adult rats with intact pituitaries receiving daily subcutaneous injections of 100 I.U. human chorionic gonadotropin for 5 days and from control animals. After human chorionic gonadotropin administration testosterone concentrations were increased in total homogenates of whole testis tissue, interstitial tissue and seminiferous tubules. The testosterone production from endogenous precursors was enhanced only in total homogenates of whole testis tissue and interstitial tissue obtained from testes of human chorionic gonadotropin-treated rats. The production of testosterone in the corresponding homogenates of isolated seminiferous tubules was very low. The specific activity of 3 beta-hydroxysteroid dehydrogenase was increased in total homogenates of whole testis tissue, isolated interstitial tissue and seminiferous tubules. No effect was observed on the specific activities of marker enzymes such as cytochrome c oxidase, monoamine oxidase, steroid sulfatase and lactate dehydrogenase, whereas the specific activities of carboxyl esterase were decreased in homogenates of whole testis tissue and interstitial tissue. Total activity of monoamine oxidase was increased in homogenates of interstitial tissue of tests from human chorionic gonadotropin treated rats. After the same prolonged human chorionic gonadotropin treatment the concentration of pregnenolone was increased in mitochondrial fractions of whole testis tissue, interstitial tissue and seminiferous tubules, and the amount of protein isolated in the mitochondrial fraction of interstitial tissue increased by 40%. Steroid production (estimated as pregnenolone) from endogenous precusors by mitochondrial fractions of whole testis tissue and interstitial tissue were increased after human chorionic gonadotropin treatment, for whole testis from 580 pmol/mg mitochondrial protein per h to 1420 pmol/mg per h; and for interstitial tissue from 2665 pmol/mg per h to 7050 pmol/mg per h. The production of pregnenolone in mitochondrial fractions obtaine from isolated seminiferous tubules was very low and contributed hardly at all to the total pregnenolone production in mitochondrial fractions of whole testis tissue from normal rats as well as from human chorionic gonadotropin-treated rats.     

Plasticity of allogeneic muscle tissue implanted into a traumatized rat muscle on exposure to He-Ne laser light

The combined effect of muscle tissue alloplasty and He-Ne laser light on the regeneration of a traumatized skeletal muscle was studied. In implantation of untreated allogeneic tissue into the region of transverse cutting of the gastrocnemius muscle exposed to laser light in vivo before surgery, the muscle regeneration in recipient rats was successful. Active regeneration of the allogeneic tissue and the muscle tissue of the recipient rat was noted. In implantation of laser-exposed allogeneic tissue into the region of trauma of unexposed gastrocnemius muscle, the muscle regeneration was less successful. The alloplastic implant was replaced by connective tissue and prevented the growth of the muscle tissue of the recipient rat in the defective zone. Implantation of laser-exposed allogeneic muscle tissue into the region of trauma of the muscle x-irradiated to a dose of 20 Gy enhanced destructive processes in it.     

Influence of tissue preparation techniques on p53 expression in bronchial and bladder carcinomas, assessed by immunofluorescence staining and flow cytometry.

In a series of bronchial and bladder carcinomas, p53 protein expression was examined. Samples from formalin-fixed, paraffin-embedded tissue (routine-treated) were compared with parallel samples of fresh tissue and tissue fixed in paraformaldehyde and ethanol. The expression of p53 was measured by immunofluorescence staining and dual parameter flow cytometry, with simultaneous monitoring of DNA content. For each tumor, p53 fluorescence with different fixatives was expressed relative to fresh tissue. The p53 fluorescence signals were on average brighter from routine-treated tissue than from fresh tissue. The tissue fixed in paraformaldehyde showed no difference from fresh tissue. In the ethanol-fixed tissue, however, fluorescence signals were reduced by nearly 70%, and the fraction of detectable p53 positive cells in tumor tissue was reduced by more than 50%. This loss of fluorescence was probably due to a leakage of the antigen from nucleus to cytoplasm. Pepsin treatment did not influence p53 fluorescence. Within the same tumor, the S-phase fraction in p53 positive cells was significantly higher than in p53 negative cells (13.1 +/- 2.0% vs. 6.5 +/- 0.8%). This pattern was not influenced by formalin fixation or pepsin treatment. Our study demonstrates that in measuring a nuclear antigen, tissue handling may influence the results, and care should be taken to optimize the preparation procedure. Using the antibody PAb 1801, p53 expression measured in archival material is not reduced as compared to fresh tissue.     

非小细胞肺癌组织中mir-483-5p的差异表达

目的:寻找非小细胞肺癌组织中微小RNA(miRNA)表达谱的差异,并鉴定非小细胞肺癌组织与癌旁组织差异表达的miRNA。方法:应用miRNA芯片分别检测非小细胞肺癌组织与癌旁组织的miRNA表达丰度,并比较两者中差异表达的miRNA;挑选mir-483-5p通过荧光定量PCR进行验证。结果:非小细胞肺癌组织与癌旁组织检测的miRNA表达谱存在较大差异。其中,鳞癌组织中有16条miRNA上调,22条miRNA下调;腺癌组织中有40条miRNA上调,51条miRNA下调。对mir-483-5p进行了定量PCR验证,发现其在非小细胞肺癌组织中的表达丰度显著高于癌旁的正常组织。结论:mir-483-5p在非小细胞肺癌组织中高表达。     

Haptoglobin release by human adipose tissue in primary culture

Haptoglobin is a putative adiposity marker because its concentration in blood is increased in obese humans. The present studies examined haptoglobin release by explants of adipose tissue in primary culture. Haptoglobin was released by explants of human visceral and subcutaneous adipose tissue at a nearly linear rate over 48 h. Explants of visceral adipose tissue released more haptoglobin than did explants of subcutaneous adipose tissue. The release of haptoglobin was quite variable, but there was a close correlation between haptoglobin release by visceral adipose tissue and that by explants of subcutaneous tissue from the same individual. Dexamethasone and niflumic acid, a cyclooxygenase-2 inhibitor, both inhibited haptoglobin release. There was release of haptoglobin by both isolated adipocytes and the adipose tissue matrix remaining after collagenase digestion of human adipose tissue. However, the amount of haptoglobin released by human adipose tissue explants in primary culture was quite low in relationship to the circulating level of haptoglobin.     

Androgen metabolism in male and female breast tissue

Incubation studies have been carried out using normal breast tissue and breast tissue from patients with gynecomastia, mammary dysplasia and breast carcinoma to determine the pattern of androstenedione metabolism. All tissues formed estrone (E₁) and testosterone (T) in all incubations. Estradiol (E₂) was isolated in incubations of tissue from 1 to 6 patients with mammary dysplasia, 5 of 6 patients with gynecomastia and in all incubations with normal and carcinoma tissue. Estrone formation was lowest in mammary dysplasia and gynecomastia, and higher in apparently normal breast tissue. The greatest E₁ formation was found in incubations with breast carcinoma tissue, although there was considerable variation within this tissue group. Estradiol formation was low in all tissues, with the highest conversion rates in carcinoma tissue. Testosterone formation in carcinoma tissue was greater than in mammary dysplasia or gynecomastia, but similar to apparently normal tissue. These results indicate that breast tissue from different pathological states varies in its capacity to aromatize androstenedione (A) to estrogenic products and to convert it to other androgens. They have also shown that the pattern of metabolism is distinctive for the nature of the pathological abnormality.     

云南秋海棠属植物叶片横切面比较解剖研究

报道30种主产于云南的秋海棠属植物叶片的横切面解剖构造特征。采用常规石蜡切片法切片观察,结果表明:云南秋海棠属植物叶片薄、横切面均为异面叶、呈典型的阴叶结构,叶肉组织虽有栅栏组织和海绵组织的分化,但栅栏组织不发达,占叶肉组织的比例较小。表皮多为单表皮,极稀复表皮,表皮毛均由多细胞组成。气孔集中于下表皮,孔下室或下陷气孔特大、通气组织极发达;角质层形状多样,呈均匀增厚、瘤状和片状突起;叶绿体椭球形、数多、个体大,主要分布于叶肉组织,集中于栅栏组织。解剖构造特征在各分类组内呈现不完全一致性,而在相同茎的形态类型中有些较一致的特征,在不同种间解剖特征各有差别;根状茎和直立茎类型种类的横切面组织结构表现为表皮细胞壁外的角质层薄、栅栏组织与叶肉组织厚度比例较小等弱光照、湿生等适应性较弱的特征。球茎类型的种类表现为角质层较厚、栅栏叶肉组织厚度比例较大等适应略为干燥和较强光照的特征。     

Biomechanical effect of rapid mucoperiosteal palatal tissue expansion with the use of osmotic expanders

The comparative study was performed to investigate the biomechanical properties (maximum tangential stiffness, maximum tangential modulus and tensile strength) of expanded mucoperiosteal palatal tissue after rapid expansion regimen correlated with histological findings. Rabbit palatal model was used to correlate the non-operated control group, sham-operated control (subperiosteal tissue dissection) groups and 24- and 48-hour tissue expansion groups. There was no observed damage of tissue collagen network in both tissue expansion groups analyzed immediately after expansion, and biomechanical profile was not significantly different from the profile of control groups. However, rapid tissue expansion activates remodeling of mucoperiosteal tissue structure that revealed significant changes in mechanical properties during the 4-week follow-up. The 24-hour expansion induced transient increase of resilience observed 2 weeks after surgery in comparison to the control groups. As a result of maturation of newly created collagen fibers and mucoperiosteum rebuilding, there were no significant differences between any of the analyzed tensile parameters 4 weeks after the 24-hour expansion. Increased and elongated inflammatory response and connective matrix synthesis observed during healing of 48-hour expanded tissue led to a significant decrease of tensile strength value in comparison to the control groups. Even though 4 weeks after surgery, the resilience of 48-hour expanded tissue was similar to the control groups, tissue healing was not completed and limited scar formation might considerably change the final biomechanical tissue profile. These findings provide new information about tensile properties to rapid mucoperiosteal palatal tissue expansion with the use of osmotic expanders for cleft palate repair by tissue augmentation.     

降胆固醇乳酸菌对肉鸡胆固醇的影响研究

目的探讨经体外实验筛选出的降胆固醇乳酸菌对肉鸡血清胆固醇及胸肌组织、腿肌组织和肝脏组织胆固醇含量的影响。方法将肉鸡随机分为2组,对照组喂食普通饲料,实验组喂食含有DM86056菌的饲料;喂食56d后分离其血清及肝脏等组织,并应用硫酸铁铵法测定其胆固醇含量,观察对照组与实验组结果差异是否有显著性。结果对肉鸡血清、胸肌组织、腿肌组织胆固醇含量测定后,对照组与实验组结果差异有显著性（P〈0．05）,而对肝脏组织胆固醇含量测定后,对照组与实验组结果差异无显著性（P〉0．1）。结论菌株DM86056可明显降低肉鸡血清、胸肌组织、腿肌组织中胆固醇含量,而对肝脏组织胆固醇含量没有明显改变。     

Continuous versus intraoperative expansion in the pig model.

Continuous tissue expansion utilizing a continuous infusion device that maintains a constant expander pressure was previously demonstrated to be feasible and successful in obtaining rapid tissue expansion in a canine model. Intraoperative tissue expansion has been described and has gained some clinical acceptance as a method to gain rapid expansion. We compared the efficacy of continuous tissue expansion versus intraoperative tissue expansion in a piglet model. After completing a pilot study, continuous tissue expansion was performed in six pigs (14.5 to 20 kg) on one flank over a 3-day period utilizing an improved prototype device; at the termination of continuous tissue expansion, intraoperative tissue expansion was performed on the opposite flank. There were no complications or continuous tissue expansion device malfunctions. Intraoperative tissue expansion gave a true gain in area of 7.4 percent, while continuous tissue expansion produced a 22 percent gain (p < 0.02). When the effects of both recruitment and expansion were added, continuous tissue expansion gave a dividend of 286 percent versus 192 percent for intraoperative tissue expansion (p < 0.01). Biomechanically, intraoperative tissue expansion skin showed few differences from unexpanded skin, while continuous tissue expansion skin showed a significant increase in stress relaxation (47.78 versus 38.74) and decrease in breaking strength. Histologic analysis revealed some epidermal hyperplasia and inflammation surrounding the continuous tissue expansion expander and some vascular congestion over the intraoperative tissue expansion expander. We conclude that continuous tissue expansion is superior to intraoperative tissue expansion and that the prototype device may be useful clinically.     

Estrone sulfate and sulfatase activity in human breast cancer and endometrial cancer

Estrone sulfate (E1-S) in the serum and tissues of patients with breast cancer or endometrial cancer was measured by a direct radioimmunoassay without hydrolysis. The concentration of E1-S in breast cancer tissue was 1.64 +/- 0.28 ng/g wet wt (+/- SE), lower than in surrounding normal breast tissue (4.46 +/- 1.23). Estradiol-17 beta(E2)/E1-S was higher in endometrial cancer tissue than normal endometrial tissue. Estrone sulfatase activity in breast cancer tissue was 0.81 +/- 0.23 nmol/h/mg protein, higher than in surrounding normal breast tissue (0.35 +/- 0.11). These results suggest that E1-S, which is abundant in the peripheral circulation, is hydrolyzed by sulfatase in breast cancer tissue or endometrial cancer tissue and liberates free estrogens, which may stimulate the growth of these malignant tumors.     

Ontogeny of American paddlefish lymphoid tissues

The temporal and spatial distribution of American paddlefish Polyodon spathula immune cell populations was determined using enzyme cytochemistry and immunohistochemistry. Monocytes and macrophages were present in the renal haematopoietic tissue, spleen, meningeal myeloid tissue, cardiac myeloid tissue and lamina propria of the spiral valve at 7 days post-hatch (dph). Immature lymphocytes were present in the renal haematopoietic tissue, spleen, meningeal myeloid tissue, cardiac myeloid tissue, thymus and lamina propria of the spiral valve at 7 dph. Type A lymphocytes (T-cell like) were demonstrated in the thymus by 21 dph. Type B immunoglobulin positive lymphocytes (B-cell like) were present in the renal haematopoietic tissue, cardiac myeloid tissue and lamina propria of the spiral valve by 7 dph, the thymus at 21 dph, the spleen by 56 dph, and were not observed in the meningeal myeloid tissue of paddlefish aged 7–28 dph. Granulocytes were present in the renal haematopoietic tissue, thymus, spleen, meningeal myeloid tissue, cardiac myeloid tissue and lamina propria of the spiral valve by 7 dph. The spleens in 7–28 dph fish were predominately red pulp. Differentiation of leukocytic and erythrocytic compartments (white and red pulp, respectively) was not apparent in the spleen until 56 dph. Remarkable thymic cortical and medullary differentiation was not yet present at 28 dph, and the thymus was not sampled at 56, 84 or 112 dph. The cardiac myeloid tissue was not developed until 56 dph. Peyer's patches were present in the lamina propria by 56 dph. Paddlefish lympho-myeloid structures are therefore slow to develop, and vaccination procedures should be performed at 2–4 months post-hatch.