Time dependent study to evaluate the efficacy of zinc on hepatic marker enzymes and elemental profile in serum and liver of protein deficient rats

This study was designed to determine the time dependent protective effects of zinc sulfate on the serum and liver marker enzymes along with elemental profile in protein deficient Sprauge Dawley (S.D.) female rats. Zinc sulfate in the dose of 227 mg/l in drinking water was administrated to normal control as well as protein deficient rats for a total duration of 8 weeks. The effects of different treatments were studied on enzymes like alkaline phosphatase (ALP), aspartate aminotransferases (AST) and alanine aminotransferases (ALT) in rat serum at different time intervals of 1, 2, 4 and 8 weeks and in the rat liver at the end of study. The status of different essential elements in liver was also studied. The serum ALP activity got significantly depressed when estimated at the intervals of 4 and 8 weeks. Activity of serum ALT was significantly increased after 4 weeks interval in protein deficient rats and the increasing trend continued upto 8 weeks of protein deficiency. On the other hand, activity of AST showed a significant increase just after 2 weeks and activity continued to be increased up to 8 weeks. Moreover activities of all the hepato marker enzymes showed a significant increase in liver of protein deficient rats. Interestingly, supplementation of Zn to protein deficient rats helped in regulating the altered activities of ALP, AST and ALT both in serum and liver. However, zinc treatment alone to normal rats did not indicate any significant change in the activities of all the enzymes in liver as well serum except at the interval of 2 weeks where a marginal increase in the activity of AST was seen. It has also been observed that concentrations of zinc, copper, iron and selenium were found to be decreased significantly in protein deficient animals. However, the levels of these elements came back to within normal limits when zinc was administrated to protein deficient rats. Published online December 2004     

凝集素亲和层析研究人血清肝型碱性磷酸酶的糖链结构

用麦胚凝集素(WGA)将人血清肝型和骨型碱性磷酸酶(ALP)分离,再将肝型ALP通过蔓陀萝凝集素(DSA)亲和层析柱作醋酸梯度洗脱,发现正常人、急性肝炎、慢性肝炎、肝硬化和原发性肝癌各组各自的混合血清ALP的层析行为有明显差异。正常人只有不结合组分,而各种良性肝病出现数目不同(2—3个)的弱结合组分,肝癌则还出现3个強结合组分,各组分ALP在不同肝病血清中的出峰时间基本恒定。这种ALP洗脱峰的不均一性在单个病人血清中依然存在,是ALP糖链结构不同而引起的。由此推测肝病时ALP上N糖链的天线数增加,肝病越发展成慢性或恶性,血清ALP和DSA的亲和力也越強。这些结果有可能在临床上鉴别各类不同的肝病。     

Effect of cyclic AMP-dependent phosphorylation on the reactivity of sulphydryl groups in cardiac sarcoplasmic reticulum.

1. The metabolism of 4-ethynylbiphenyl has been studied in vitro with subcellular fractions of normal and induced rat liver, and rat intestinal microflora (caecal contents). 2. Oxidation was NADPH-dependent, was inhibited by CO and stimulated by pretreatment with phenobarbitone or 3-methylcholanthrene. 3. Oxidation of the ethynyl group occurred in washed microsomal preparations, but not significantly in soluble fractions. Oxidation of the ethynyl group by a microsomal fraction preceded aromatic hydroxylation and no metabolites containing the intact ethynyl group were detected. 4. The major metabolite in liver fractions was biphenyl-4-ylacetic acid. This was the only product produced by a modified Udenfriend system. 5. Metabolism of 4-ethynylbiphenyl by rat caecal contents under anaerobic conditions produced very small amounts of 4-vinylbiphenyl. 6. In a modified Ames test with Salmonella typhimurium TA98, 4-ethynylbiphenyl gave a weak positive result that was doubled after 'activation' with an induced rat S9 fraction.     

Gamma-glutamyltranspeptidase isoenzyme forms and lipoproteins in normal and pathological sera

The molecular nature of serum (from normal subjects and from patients affected by various hepatobiliary diseases) gamma-glutamyltranspeptidase (GGT) isoenzymes has been studied by selective lipoprotein precipitation. Some fractions co-precipitate with LDL + VLDL (pre-beta-, beta-, beta/gamma-, gamma-, and dep-GGT fractions) or with HDL (partial precipitation of alpha 1-GGT in cirrhosis). Alpha 1-GGT + alpha 2-GGT in normal subjects, and Alb-GGT did not precipitate with either of the precipitation treatments. Total GGT and its isoenzymes were stable at 4 degrees C and at -20 degrees C for at least 20 days, with the exception of Alb-GGT which at -20 degrees C decreased by 20%. The percentage of GGT associated with LDL + VLDL appeared to be a possible marker to discriminate liver tumors from cirrhosis. A cut-off value of 20 U/L of this marker yielded a diagnostic sensitivity of 87% and a diagnostic specificity of 85%.     

序列固相凝集素亲和层析法研究大鼠诱发肝癌中碱性磷酸酶ALP的糖链变化

 凝集素是一种能同复合糖中不同糖基序列特异性相结合的糖结合蛋白质。本文在研究二乙基亚硝胺诱发肝癌中碱性磷酸酶(ALP)活力和同工酶的基础上,利用四种固相凝集素柱组成的序列亲和层析,配合外切糖苷酶β-N-酰氨基葡萄糖苷酶,研究了诱发肝癌ALP糖链中“Bisecting”GlcNAc残基的变化。研究结果表明:(1)大鼠诱发肝癌ALP的活性较正常肝脏明显增高,其性质和正常肝脏ALP基本相同,两者均属于组织非特异性ALP,提示大鼠肝癌发生过程中没有同工酶谱的变化。(2)大鼠诱发肝癌ALP糖链的“Bisecting”GlcNAc残基含量较正常鼠肝明显增加。     

The pathophysiology of human obstructive cholestasis is mimicked in cholestatic Gold Syrian hamsters

Obstructive cholestasis causes liver injury via accumulation of toxic bile acids (BAs). Therapeutic options for cholestatic liver disease are limited, partially because the available murine disease models lack translational value. Profiling of time-related changes following bile duct ligation (BDL) in Gold Syrian hamsters revealed a biochemical response similar to cholestatic patients in terms of BA pool composition, alterations in hepatocyte BA transport and signaling, suppression of BA production, and adapted BA metabolism. Hamsters tolerated cholestasis well for up to 28 days and progressed relatively slowly to fibrotic liver injury. Hepatocellular necrosis was absent, which coincided with preserved intrahepatic energy levels and only mild oxidative stress. The histological response to cholestasis in hamsters was similar to the changes seen in 17 patients with prolonged obstructive cholestasis caused by cholangiocarcinoma. Hamsters moreover upregulated hepatic fibroblast growth factor 15 (Fgf15) expression in response to BDL, which is a cytoprotective adaptation to cholestasis that hitherto had only been documented in cholestatic human livers. Hamster models should therefore be added to the repertoire of animal models used to study the pathophysiology of cholestatic liver disease.     

Changes of glycoconjugates in human hepatocellular carcinoma

Receptors of 12 lectins in 25 cases of human hepatocellular carcinomas (HCC) were histochemically investigated by avidin-biotin-peroxidase complex (ABC) method. Liver tissues of five cirrhotic patients and five normal subjects were used as controls. SJA receptor was absent both in HCC and controls, while LCA and PSA receptors were present in all tissues studied here. Receptors of DBA, PHA, PNA, UEAI and SBA which did not bind to normal, cirrhotic and pericarcinomatous liver tissues had the positive rates of 4%, 44%, 16%, 4% and 12% in HCC, respectively. Four lectins which strongly bound to the non-cancer liver tissues had their receptors in 96% (ConA, WGA, RCAI) and 36% (BSAI) of HCC. The pretreatment of tissue sections with neuraminidase abolished most of WGA receptors and exposed some PNA binding sites. There were many differences in lectin distribution between HCC and noncancer liver tissues. The changes of glycoconjugates in HCC were discussed.     

Comparative evaluation of the glycosaminoglycan composition in plasma membranes of normal and transformed mouse liver cells

Using electrophoresis in agarose gel, a comparative study of composition of membrane-bound glycosaminoglycans (GAG) from normal mouse liver cells and from o-aminoazatoluene-induced mouse hepatoma cells was carried out. Differences in the composition and localization of GAG were revealed. The plasma membrane fraction of hepatoma cells contained no GAG; the bulk of GAG (approximately 98%) was localized in the plasma membrane free fraction. Within this fraction GAG contained no heparan sulfates with a high electrophoretic mobility that were detected in plasma membranes of normal liver cells. The possible involvement of proteoglycans bound to cell surface in transmembrane signal transfer is discussed.     

S-腺苷蛋氨酸治疗胆汁淤积性肝病伴抑郁／焦虑的临床效果研究

目的：研究s-腺苷蛋氨酸治疗胆汁淤积性肝病伴抑郁／焦虑患者的临床效果。方法：选择2011年3月-2013年3月我院收治的51例不同病因的胆汁淤积性肝病（药物性肝损害13例、慢性乙型肝硬化14例、酒精性肝硬化11例、自身免疫性肝病6例、肝癌5例、胆管癌2例）并抑郁／焦虑的患者,予s-腺苷蛋氨酸1．0g治疗2周,应用SDS／SAS量表分别评估和比较治疗前后各组患者抑郁／焦虑程度的评分情况。结果：S-腺苷蛋氨酸治疗后,所有组别胆汁淤积性肝病肝病改善的临床总有效率94．12％,其中药物性肝损害、慢性乙型肝硬化、酒精性肝硬化、自身免疫性肝硬化总有效率均为100．00％,肝癌的有效率为60．00％,胆管癌的有效率为50．00％,药物性肝损害患者临床疗效与其他各组有差异（P〈0．05）;药物性肝病患者SDS和SAS评分均较治疗前显著降低（P〈0．05）。而慢性乙型肝硬化、酒精性肝硬化、自身免疫性肝病、肝癌、胆管癌患者SDS和SAS评分与治疗前相比均无统计学差异（P〉0．05）。结论：S-腺苷蛋氨酸可改善药物性胆汁淤积性肝病并轻、中度抑郁／焦虑患者的肝功能,并有效减轻其抑郁／焦虑情绪。     

S-腺苷蛋氨酸治疗胆汁淤积性肝病伴抑郁/焦虑的临床效果研究

目的:研究S-腺苷蛋氨酸治疗胆汁淤积性肝病伴抑郁/焦虑患者的临床效果。方法:选择2011年3月~2013年3月我院收治的51例不同病因的胆汁淤积性肝病(药物性肝损害13例、慢性乙型肝硬化14例、酒精性肝硬化11例、自身免疫性肝病6例、肝癌5例、胆管癌2例)并抑郁/焦虑的患者,予S-腺苷蛋氨酸1.0 g治疗2周,应用SDS/SAS量表分别评估和比较治疗前后各组患者抑郁/焦虑程度的评分情况。结果:S-腺苷蛋氨酸治疗后,所有组别胆汁淤积性肝病肝病改善的临床总有效率94.12%,其中药物性肝损害、慢性乙型肝硬化、酒精性肝硬化、自身免疫性肝硬化总有效率均为100.00%,肝癌的有效率为60.00%,胆管癌的有效率为50.00%,药物性肝损害患者临床疗效与其他各组有差异(P0.05);药物性肝病患者SDS和SAS评分均较治疗前显著降低(P0.05)。而慢性乙型肝硬化、酒精性肝硬化、自身免疫性肝病、肝癌、胆管癌患者SDS和SAS评分与治疗前相比均无统计学差异(P0.05)。结论:S-腺苷蛋氨酸可改善药物性胆汁淤积性肝病并轻、中度抑郁/焦虑患者的肝功能,并有效减轻其抑郁/焦虑情绪。     

A simple procedure for the isolation of highly purified lysosomes from normal rat liver

A procedure for the isolation of highly purified lysosomes from normal rat liver is described. The method depends on the swelling of mitochondria when the postnuclear supernatant fraction is incubated with 1 mM Ca2+. The lysosomes can then be separated from the swollen mitochondria by Percoll density gradient centrifugation. The lysosomal fraction obtained by our method was enriched more than 120-fold in terms of the marker enzymes with a yield of 25%. The electron microscopic examination and the measurement of the activities of marker enzymes for various subcellular organelles indicated that our lysosomal preparation was essentially free from contamination by other organelles.     

Identification of liver-intestine cadherin in hepatocellular carcinoma--a potential disease marker

Liver-intestine cadherin (LI-cad) is a non-classical cadherin, which is expressed during intestinal development, but absent in normal liver tissue. Our earlier investigation has detected overexpression of LI-cad in gastric adenocarcinoma and indicated its association with lymph node metastasis. Herein, we found in RT-PCR and TaqMan Q-PCR that LI-cad was identified in HCC cell lines, HuH-7, Hep-3B, and PLC/PRF/5, but not in MIHA and HepG2 non-tumorigenic cells. Immunofluorescence cytochemistry assay revealed that the LI-cad was predominantly expressed in cytoplasm of HCC cells, contrary to that of E-cad immunostain at the plasma membrane region. By testing against 18 pairs of HCC and adjacent non-tumor tissues, 13 cases (72.2%) showed over expression of LI-cad in HCC tissues, 2 cases (11.1%) were similar, and 3 cases did not yield detectable signal. None of the 6 normal liver specimens tested was positive with LI-cad. Taken together, LI-cad could be a potential disease marker for HCC.     

Lipid peroxidation and physical properties of smooth and rough hepatoma microsomes

Smooth and rough endoplasmic reticulum of two Morris hepatomas, the slow growing 9618A and the fast growing 3924A, have been isolated, and their biochemical composition, supramolecular organization, and response to the action of peroxidative agents have been studied. Cytochrome P450 content and lipid availability are the limiting factors of their peroxidizability. The hemoprotein content is reduced about 80% in hepatoma 9618A and is virtually absent in hepatoma 3924A. The peroxidizability decreases with increasing growth rate of the tumor. The protein, phospholipid, and cholesterol content, the fatty acid composition as well as the double bond index, and the saturated and unsaturated fatty acid content are reported. Differences have been found between normal liver and tumors and between the fractions within a given tumoral tissue. The molecular order, as determined by fluorescence anisotrophy decay of DPH, increases in total microsomes and in the smooth fraction going from liver 9618A to 3924A, whereas for the rough fraction it is the same in liver and hepatoma 9618A; in 3924A it increases of about 30%. Fluidity decreases in total microsomes going from liver to 3924A, to 9618A. In both the purified fractions it decreases with increasing deviation of the tumor.     

Role of zinc in regulating the levels of hepatic elements following nickel toxicity in rats

This study was designed to determine the protective effects of zinc on the hepatotoxicity induced by nickel in rats. Female Sprague-Dawley (SD) rats received either nickel sulfate alone in the dose of 800 mg/L nickel in drinking water, zinc sulfate alone in the dose of 227 mg/L zinc in drinking water, and nickel plus zinc or drinking water alone for a total duration of 8 wk. The effects of different treatments were studied on activities of rat liver marker enzymes like alkaline phosphatase (ALP), alanine aminotransferase (ALT), and aspartate aminotransferases (AST) and on the status of essential elements in rat liver. The study revealed a significant increase in the activities of enzymes ALP and ALT in rats subjected to nickel treatment. Interestingly, zinc supplementation to rats treated with nickel brought back the raised activities of these enzymes to within normal limits. Further, the levels of elements in liver that include zinc, copper, selenium, and potassium were found to be significantly suppressed following nickel treatment, whereas the levels of iron and sulfur were elevated. However, zinc treatment alone did not cause any appreciable change in the concentration of these elements. To the contrary, when zinc was given to nickel-treated rats, the concentrations of zinc, copper, potassium, and phosphorus were not significantly different from that of normal controls, whereas the levels of iron, selenium, and sulfur were improved in comparison to nickel-treated rats but were not within the normal limits. The present study concludes that zinc has the ability to maintain the levels of hepatic elements and has bearing in regulating the liver functions by maintaining the activities of marker enzymes in conditions of nickel toxicity.     

Levels of hepatitis B virus replicative intermediate in serum samples of chronic hepatitis B patients

Hepatitis B virus (HBV) cccDNA levels is an absolute marker of HBV replication in the liver of HBV infected patients. This study aimed to quantify the HBV cccDNA levels in sera and liver tissue samples of treatment naïve patients with chronic hepatitis B. Eighty one chronic hepatitis B (CHB) treatment naïve patients were enrolled from January 2009 to June 2011. Total HBV DNA and HBV cccDNA levels were quantified using sensitive real time PCR assay. The mean age of recruited patients was 34 ± 11.5 years. Fifty four (66.7 %) patients were HBeAg negative. Liver tissue samples were available from 2 HBeAg positive and 21 HBeAg negative CHB patients. The amount of total intrahepatic HBV DNA ranged from 0.09 to 1508.92 copies/cell. The median intrahepatic HBV cccDNA was 0.31 and 0.20 copies/cell in HBeAg positive and HBeAg negative cases, respectively. Serum HBV cccDNA was detectable in 85.2 % HBeAg positive and 48.1 % HBeAg negative CHB patients. Median serum HBV cccDNA was 46,000 and 26,350 copies/mL in HBeAg positive and HBeAg negative subjects, respectively. There was a significant positive correlation between the levels of intrahepatic total HBV DNA and intrahepatic HBV cccDNA (r = 0.533, p = 0.009). A positive correlation was also seen between serum HBV cccDNA levels and serum HBV DNA levels (r = 0.871, p < 0.001). It was concluded that serum HBV cccDNA could be detectable in higher proportion of HBeAg positive patients compared to HBeAg negative patients. Moreover, the median level of serum HBV cccDNA was significantly higher in HBeAg positive patients in contrast to HBeAg negative subjects.     

Lipid peroxidation and physical properties of smooth and rough hepatoma microsomes

Smooth and rough endoplasmic reticulum of two Morris hepatomas, the slow growing 9618A and the fast growing 3924A, have been isolated, and their biochemical composition, supramolecular organization, and response to the action of peroxidative agents have been studied. Cytochrome P₄₅₀ content and lipid availability are the limiting factors of their peroxidizability. The hemoprotein content is reduced about 80% in hepatoma 9618A and is virtually absent in hepatoma 3924A. The peroxidizability decreases with increasing growth rate of the tumor. The protein, phospholipid, and cholesterol content, the fatty acid composition as well as the double bond index, and the saturated and unsaturated fatty acid content are reported. Differences have been found between normal liver and tumors and between the fractions within a given tumoral tissue. The molecular order, as determined by fluorescence anisotrophy decay of DPH, increases in total microsomes and in the smooth fraction going from liver 9618A to 3924A, whereas for the rough fraction it is the same in liver and hepatoma 9618A; in 3924A it increases of about 30%. Fluidity decreases in total microsomes going from liver to 3924A, to 9618A. In both the purified fractions it decreases with increasing deviation of the tumor.     

Value of fine needle aspiration in the diagnosis of breast lesions.

OBJECTIVE: To evaluate the accuracy values of 276 fine needle aspriations (FNA) of breast lesions with a subsequent excisional biopsy diagnosis and to make a comparison between 25 studies of the literature using the same criteria to calculate those values. STUDY DESIGN: Cytologic findings were compared with the histologic diagnosis of each mass. The correlation of results was analyzed by a decision-analysis approach, and the following values concerning diagnostic accuracy were calculated in the present study and in 25 other reports: sensitivity, specificity, positive predictive value, negative predictive value, false positive fraction and false negative fraction. To calculate those values, we eliminated unsatisfactory results and assumed that suspicious and positive cytologic findings represented carcinoma of the breast. RESULTS: Comparing our results with the means in the literature (numbers in parenthesis), FNA detected cancer with a sensitivity of 92.1% (87.7%), specificity of 98.6% (94.7%), positive predictive value of 99.4% (92.8%), negative predictive value of 82.1% (90.7%), false positive fraction of 0.6% (7.1%) and false negative fraction of 17.9% (13.4%); in 6.2% of cases the material was unsatisfactory (13.4%). CONCLUSION: All the rates varied enormously between the studies and during the past 13 years. It seems that false positive and false negative fractions tended to diminish and stabilize in more recent years, and specificity and sensitivity underwent a slight increase. The differences between the rates of those studies suggest that FNA of the breast has some unavoidable limitations.     

Changes of glycoconjugates in human hepatocellular carcinoma

Summary Receptors of 12 lectins in 25 cases of human hepatocellular carcinomas (HCC) were histochemically investigated by avidin-biotin-peroxidase complex (ABC) methol. Liver tissues of five cirrhotic patients and five normal subjects were used as controls. SJA receptor was absent both in HCC and controls, while LCA and PSA receptors were present in all tissues studied here. Receptors of DBA, PHA, PNA, UEA_I and SBA which did not bind to normal, cirrhotic and pericarcinomatous liver tissues had the positive rates of 4%, 44%, 16%, 4% and 12% in HCC, respectively. Four lectins which strongly bound to the non-cancer liver tissues had their receptors in 96% (ConA, WGA, RCA_I) and 36% (BSA_I) of HCC. The pretreatment of tissue sections with neuraminidase abolished most of WGA receptors and exposed some PNA binding sites. There were many differences in lectin distribution between HCC and noncancer liver tissues. The changes of glycoconjugates in HCC were discussed.     

Role of the cytomegalovirus (CMV)-antigenemia assay as a predictive and follow-up detection tool for CMV disease in AIDS patients

Forty-two patients were evaluated to determine the value of the CMV antigenemia (CMV-Ag) test as a follow-up marker as well as a prediction marker of CMV disease. Twenty patients were positive for at least one positive CMV-Ag assay and 9 of them developed CMV retinitis. With the threshold value (10 positive cells), sensitivity was 56% and specificity was 94%. The CMV-Ag assay, with the threshold value, produced high specificity, positive predictive value and negative predictive value but relatively poor sensitivity. Eight patients experienced CMV disease relapse a total of 16 times. At relapse, 8 of the 16 times showed negative for CMV-Ag assay; 7 underwent systemic maintenance while 1 underwent local maintenance. It is inferred that the CMV-Ag test is a poor follow-up marker to detect the relapse of CMV disease, particularly in patients undergoing systemic maintenance.