Hepatic expression of the proliferative marker Ki-67 and cell cycle p53 protein in chronic hepatitis C [a histopathological and immunohistochemical study]

To evaluate hepatic expression of the nuclear proliferative marker Ki-67 and the cell cycle marker p53 oncoprotein in chronic hepatitis C in relation to the advanced stages of liver fibrosis in HCV positive Egyptian patients. Paraffin-embedded liver biopsy specimens were studied from 21 untreated patients with chronic HCV infection. All patients were HCV antibody positive, as determined by a commercially available enzyme-linked immunosorbent assay kit. Patients having other etiologies for chronic liver disease including HBV infection were not included in this study. Liver biopsies were obtained percutaneous. All biopsies were fixed in formalin, embedded in paraffin, and sectioned by microtome with a thickness of 5 ?m. Routine specimen processing involved staining slides with hematoxylin and eosin [5 levels], Masson's trichrome stain [5 levels], for a total of 10 levels per specimen All levels were screened. All specimens were examined by two pathologists, and classified by consensus for all abnormal histological findings. The histological activity index [or histological grade] was determined using Ishak grading scheme22 expressed as a semiquantitative score for portal inflammation [0-4], lobular activity sporadic lytic foci [0-4] and parenchymal confluent necrosis [0-6], and piecemeal necrosis[0-4]. The extent of fibrosis [or histological stage] was determined using Ishak score [0-6]. Steatosis was scored according to Keliner et al 2005, from grade 0 to 3; where S0 = no steatosis or less than 5% [low or medium power evaluation] of parenchymal involvement by fatty changes, S1 [mild] = 5%-33%, S2 [moderate] = >33%-66% and S3 [severe] > 66% of the hepatocytes are involved by fatty changes. Expression of p53 and Ki67 were determined by immunohistochemistry, using avidin-biotin-peroxidase. Liver histology: The studied group [n = 21] involved 16 males and 5 females [male to female ratio 3.3:1]. The histopathological findings of HCV infection, including portal lymphoid infiltration, periportal piecemeal necrosis, lymphocyte infiltration of the lobules, hepatocellular necrosis, steatosis and fibrosis, were studies. The age ranged from 31 to 59 years old with mean of 44.86 +/- 8.74, males 76.2%, females 23.8% .P53 expression was positive in 52.4% and negative in 47.6%. cytoplasmic localization dominated over nuclear expression. Ki 67 was negative in 81% of cases and positive in19% of cases, all cases in stage 6 were positive for p53 while there were no difference in the other stages of fibrosis, and this relation was statistically significant. There was no relation between the grade of necro-inflammation and the expression of p53, and this result was statistically non significant.There was a relation between the percent of steatosis and the expression of p53 as percent of positivity increases with the increase of the percent of steatosis, and this result was statistically significant using independent sample t test and regression test.All negative cases for P53 have negative Ki67 but this rule is not applied on positive cases for P53, and this relation was not statistically significant. There was no relation between the grade of necro-inflammation and the expression of Ki67, and this result was statistically non significant. Hepatic expression of the nuclear proliferative marker Ki-67 and the cell cycle marker p53 oncoprotein in chronic hepatitis C in relation to advanced stages of liver fibrosis in HCV positive patients are expressed in a considerable present of cases which should be cansdidates for follow up for early detection of hepatocellular carcinoma

Effect of androgen and schistosoniicide in murine schistomiasis mansoni: an experimental study

Impotence is aconsistent inability to sustain an erection sufficient for sexual intercourse. Testosterone administration in men with liver cirrhosis improves the sense of well-being, increase serum proteins and reduces edema without serious adverse effects. Oral, alkylated forms of testosterone can create a situation of liver toxicity. There is little evidence that other methods of administration cause liver dysfunction. Most doctors be indecisive on prescribing androgen preparations in patients with liver disease, so this work was designed to study the effect of androgen replacement [injectable form] on the murine diseased liver, and subsequently whether it can be used safely in men with chronic liver disease or not. To evaluate the effect of exogenous injectable androgen and praziquantel on the diseased liver of mice. National Hepatology and Tropical Medicine Research Institute [NHTMRI]. Materials and methods: Forty male mouse [weighing 25-30 g] were infested subcutaneously with Schistosoma mansoni [100 cercariae/animal], and then they were divided into four groups. Mice in the first group were infected only and used as infected control group. Mice of group II and IV were given the Schistosomicide, praziquantel in a dose of 0.3 mg/mouse. Androgen [Sustarion] was injected intramuscularly in a "dose of 0.125 mg/mouse, [three doses, 3 weeks apart] in group III and IV. At the end of the trial all animals were then sacrificed to study histopathologically the possible effects of androgen on the liver tissue. Liver function tests were done in animals of group I, III, and IV, first prior to study and finally by the end of study. Results of assayed liver function tests and histopathological examination were tested for statistical significant association. there were marked elevation of the liver enzymes in mice of group IV compared to the corresponding control [p<0.01] and mice of the third group [p<0.01], which reflect deterioration of hepatic function in those mice received the antibilharzial drug praziquantel. On the other hand there was statistical difference between control group [group I] and androgen treated group III [P<0.05]. Histological examination of liver sections of mice in all groups revealed the presence of typical bilharzial granulomas. The mean diameter of bilharzial granulomas clearly dropped to 283.20 micrometer in group II compared to 392.55 in corresponding control. The difference between these two groups was statistically significant [p=0.0001]. In group III there was no statistical difference in the number of egg granulomas [P>0.05] compared to group I. There was a reduction of granulomas diameter in group III and IV [animals injected with androgen] in comparison to group I [P>0.05 and P<0.01] respectively. Also comparison between the four groups as regards the type of bilharzial granulomas, it is clearly evident that the predominant type of granulomas in the androgen treated groups is the cellular type [38% and 57.1%] in group III and IV respectively and this may reflect the possible beneficial effect of androgen on the diseased liver. Our results clearly indicate that androgen have no deleterious effects on tissues of the diseased liver and hence on liver functions. In conclusion androgen therapy [injectabi form] appears to be safe in the clinical management of erectile dysfunction patients with chronic liver disease

Study of IL-10 and Il-18 levels in relation to different stages of liver fibrosis in Egyptian chronic hepatitis C infected patients

Most acute and chronic liver diseases are characterized by inflammatory processes with enhanced expression of various pro-and anti-inflammatory cytokines in the liver. These cytokines are the driving force of many inflammatory liver disorders often resulting in fibrosis and cirrhosis. This work aimed to identify the role of serum IL-10 and IL-18 in the pathogenesis of chronic hepatitis C in different grades and stages of HCV patients and correlate their levels to the necroinflammatory grade and stage of fibrosis. A prospective study on [55] HCV infected patients 43 males and 12 females, their age ranged from 20 to 55 years with a mean of 41.3 year and [20] age and sex matched control [HCV-ve] from National Hepatology and Tropical Medicine Research Institute clinics, from whom consents were taken. The patients included in this study were those originally enrolled on protocol for treatment [interferon + ribavirin] where liver biopsy was required for eligibility of treatment according to the approved protocol. 5ml of blood was collected at the same day of biopsy for assessment of IL-10 and IL-18 levels in blood [ELISA technique]. Serum level of IL-18 was significantly higher in patients with chronic HCV [126.80 +/- 11.76 pg/ml] as compared to control group [45.97 +/- 5.26 pg/ml]; the level was even higher in cases with hepatocellular carcinoma [HCC]. IL-18 levels were significantly higher in patients with early fibrosis than in patients with advanced fibrosis., it was also higher in case with minimal to mild necro-inflammatory grade than in cases with moderate to severe grades, but that difference was not significant. As regards serum IL-10, the level was also higher in patients with chronic HCV [9.24 +/- 1.9 pg/ml] than control group [3.28 +/- 1.81 pg/ml]. Serum IL-10 level was significantly lower in early than advanced stages [P=0.04]. When classifying cases according to necro-inflammatory grade the IL-10 level was lower in minimal/mild cases than moderate /severe cases, but this difference wasn't statistically significant. Serum concentrations of IL-10 and IL18 could be correlated to the histopathological spoilage of the liver as they can predict the stage of fibrosis and hence can be used as indirect markers to assess the severity of liver disease in HCV infected patients. They can also be used as complementary markers in HCC patients

Reproductive effects of human interferon-alpha-2b administration on male albino mice testes. An experimental study

Recombinant human interferon alpha [rh-IFN-alpha] is used therapeutically in malignant disorders and chronic hepatitis. The phenotypic effects of this drug at the structural levels on testicular tissue were hardly ever addressed. Hence, this work was designed in adult male albino mice to study the phenotypic effects of rh-INF-alpha-2b on testicular tissue as well as assessing its effects on serum testosterone and gonadotropins levels. This research was planned to through light on the effects of interferon-alpha-2b [IFN-alpha-2b] on the hypothalamic-pituitary-testicular [HPT] axis of the adult male albino mice. Experimental study. National Hepatology and Tropical Medicine Research Institute [NHTMRI]. The study was conducted from November [2004] to February [2005]. Thirty sexually mature male mice were divided into three groups [10 mice in each group], namely: the control, the experimental and the recovery groups. Mice in the experimental and recovery groups were administered recombinant human interferon alpha intraperitoneally at a dose of 3000 U / mouse weekly for 12 weeks in a volume of 1.0-microliter isotonic normal' saline, then animals in the recovery group were left to recover for a further period of two months. At the end of the experiment, serum concentrations of gonadotropins and testosterone were measured and then all animals were then sacrificed to study histopathologically the possible effects of interferon on the testicular tissue. rh-IFN-alpha-2b induced remarkable decline in the serum levels of both follicle stimulating hormone [FSH] and luteinizing hormone [LH] in mice of the experimental group compared to the corresponding control and mice of recovery group. At the same time, testosterone was moderately increased in the experimental group, and then returned to its normal levels within 2 months after cessation of treatment. Histopathologically, in the experimental group, there were focal thickening of the basement membrane, degenerative changes and clumping of the germinal epithelial cells in the center of seminiferous tubules, partial desquamation of the germinal epithelium from basement membrane, reduction in the germ cell height, partial arrest of maturation and increased number of Sertoli cells. Increased number of leydig's cells and hypervascularity were detected in the interstitial spaces. In the recovery group, there was lessening of the germ cell hypoplasia manifested by restoration of spermatogenic cells and accidental disruption in the basement membrane. Most of the spermatogenic and Sertoli cells restored their polarity, height and maturation. Our results suggest that rh-INF-alpha-2b temporally affects the hypothalamic-pituitary-testicular axis [HPT], both centrally and peripherally [at the testicular level], through the lessening of FSH, LH, raise of testosterone serum levels and direct phenotypic effect on the testicular tissue

Reproductive effects of human interferon-Alpha-2b administration on male albino mice testes. An experimental study

Background: recombinant human interferon alpha [rh-IFN-alpha] is used therapeutically in malignant disorders and chronic hepatitis. The phenotypic effects of this drug at the structural levels on testicular tissue were hardly ever addressed. Hence, this work was designed in adult male albino mice to study the phenotypic effects of rh-INF-alpha-2b on testicular tissue as well as assessing its effects on serum testosterone and gonadotropins levels

Objective: this research was planned to through light on the effects of interferon-alpha-2b [IFN-alpha-2b] on the hypothalamic-pituitary-testicular [HPT] axis of the adult male albino mice

Design: experimental study

Setting: national Hepatology and Tropical Medicine Research Institute [NHTMRI]. The study was conducted from November [2004] to February [2005]

Materials and methods: thirty sexually mature male mice were divided into three groups [10 mice in each group], namely: the control, the experimental and the recovery groups. Mice in the experimental and recovery groups were administered recombinant human interferon alpha intraperitoneally at a dose of 3000 U / mouse weekly for 12 weeks in a volume of 1.0-microliter isotonic normal saline, then animals in the recovery group were left to recover for a further period of two months. At the end of the experiment, serum concentrations of gonadotropins and testosterone were measured and then all animals were then sacrificed to study histopathologically the possible effects of interferon on the testicular tissue

Results: rh-IFN-alpha-2b induced remarkable decline in the serum levels of both follicle stimulating hormone [FSH] and luteinizing hormone [LH] in mice of the experimental group compared to the corresponding control and mice of recovery group. At the same time, testosterone was moderately increased in the experimental group, and then returned to its normal levels within 2 months after cessation of treatment. Histopathologically, in the experimental group, there were focal thickening of the basement membrane, degenerative changes and clumping of the germinal epithelial cells in the center of seminiferous tubules, partial desquamation of the germinal epithelium from basement membrane, reduction in the germ cell height, partial arrest of maturation and increased number of Sertoli cells. Increased number of Leydig's cells and hypervascularity were detected in the interstitial spaces. In the recovery group, there was lessening of the germ cell hypoplasia manifested by restoration of spermatogenic cells and accidental disruption in the basement membrane. Most of the spermatogenic and Sertoli cells restored their polarity, height and maturation

Conclusion: our results suggest that rh-INF-alpha-2b temporally affects the hypothalamic-pituitary-testicular axis [HPT], both centrally and peripherally [at the testicular level], through the lessening of FSH, LH, raise of testosterone serum levels and direct phenotypic effect on the testicular tissue

Effect of anti - cox -z on tumor proliferation, angiogenesis and metastatic potential in induced colon cancer in adult albino rats: an immunohistopathological study

In this study, 60 male albino rats [average weight 100-150 g] were used. They were randomly divided into four groups, each comprised 15 rats: The first group served as a control; rats in the second and third groups were subcutaneously injected with 1,2 dimethylhydrazine [DMH] 20 mg/kg b.w. once a week for 20 weeks as well as rats in the third and fourth groups were orally administered celecoxib 20 mg/kg b.w. daily for 20 weeks. At the end of the trial, all animals were sacrificed to study the possible effects of selective cox-2 inhibitor on induced colon cancer tumor cell proliferation, angiogenesis and migration in the experimental animals. Colons, livers and lungs were taken, put in 10% neutral formalin, processed and stained for histopathological and immunohistopathological study. The tumor incidence in DMH treated group [2nd group] was 100%. Celecoxib + DMH treated group had fewer tumors and less angiogenesis compared with DMH treated group and control group [4th and 1st group]. Also, liver and lung metastasis were less in celecoxib + DMH treated group than the DMH treated group

Effect of androgen and schistosomicide in murine schistomiasis mansoni: an experimental study

Background: Impotence is a consistent inability to sustain an erection sufficient for sexual intercourse. Testosterone administration in men with liver cirrhosis improves the sense of well-being, increase serum proteins and reduces edema without serious adverse effects. Oral, alkylated forms of testosterone can create a situation of liver toxicity. There is little evidence that other methods of administration cause liver dysfunction. Most doctors be indecisive on prescribing androgen preparations in patients with liver disease, so this work was designed to study the effect of androgen replacement [injectable form] on the murine diseased liver, and subsequently whether it can be used safely in men with chronic liver disease or not

Objective: To evaluate the effect of exogenous injectable androgen and praziquantel on the diseased liver of mice

Setting: National Hepatology and Tropical Medicine Research Institute [NHTMRI]

Materials and methods: Forty male mouse [weighing 25-30 g] were infested subcutaneously with Schistosoma mansoni [100 cercariae/animal], and then they were divided into four groups. Mice in the first group were infected only and used as infected control group. Mice of group II and IV were given the Schistosomicide, praziquantel in a dose of 0.3mg/mouse. Androgen [Sustanon] was injected intramuscularly in a dose of 0.125 mg/mouse, [three doses, 3 weeks apart] in group III and IV. At the end of the trial all animals were then sacrificed to study histopathologically the possible effects of androgen on the liver tissue. Liver function tests were done in animals of group I, III, and IV, first prior to study and finally by the end of study. Results of assayed liver function tests and histopathological examination were tested for statistical significant association

Results: there were marked elevation of the liver enzymes in mice of group IV compared to the corresponding control [p<0.01] and mice of the third group [p<0.01], which reflect deterioration of hepatic function in those mice received the antibilharzial drug praziquantel. On the other hand there was statistical difference between control group [group I] and androgen treated group III [P < 0.05]. Histological examination of liver sections of mice in all groups revealed the presence of typical bilharzial granulomas. The mean diameter of bilharzial granulomas clearly dropped to 283.20 micrometer in group II compared to 392.55 in corresponding control. The difference between these two groups was statistically significant [p = 0.000].]. In group III there was no statistical difference in the number of egg granulomas [P> 0.05] compared to group I. There was a reduction of granulomas diameter in group III and IV [animals injected with androgen] in comparison to group I [P>0.05 and P<0.01] respectively. Also comparison between the four groups as regards the type of bilharzial granulomas, it is clearly evident that the predominant type of granulomas in the androgen treated groups is the cellular type [38% and 57.1%] in group III and IV respectively and this may reflect the possible beneficial effect of androgen on the diseased liver