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Microencapsulated hepatocytes and islets as in vivo bioartificial liver support system.

AIM: To confirm the xenotransplantation of microencapsulated hepatocytes and islets as a temporary bioartificial liver support system for mice with acute liver failure (ALF). METHODS: Mice were rendered ALF by a single intra-peritoneal injection of D-galactosamine (D-gal) and their tail blood was sampled to examine differences in blood ALT, albumin (ALB), total bilirubin (TB) and glucose (GLU) between 4 experimental groups. Rat hepatocytes and islets were collected and microencapsulated referring to both Sun's and Fritschy's methods. Mice were grouped into control group (CG), free hepatocyte group (FHG), microencapsulated hepatocyte group (MHG) and microencapsulated hepatocyte plus islet group (HIG). Tissue samples were subjected to microscopic and electron microscopic (EM) examinations. RESULTS: The highest survival was observed in HIG, surprisingly at 100%(16/16), while the lowest was in CG at 12.5%(2/16), with inter-group statistical difference P<0.05. ALT levels revealed no statistical difference between groups but the ALB level of HIG descended by the slightest margin (q=(0.54, 0.24, 1.33), P<0.05) at the time when it reached the lowest point in all groups. TB of HIG returned to normal reference range (NRR) statistically sooner than that of others after a fierce elevation. No statistical inter-group difference was observed in GLU levels. Fusion between hepatocytes and beta cells was demonstrated giving rise to theoretical assumptions. CONCLUSION: Hepatocytes to be microencapsulated together with islets should be a preferred in vivo hepatic functional supporting system, which can dramatically prolong survival and improve living status.

Animals↗

Protective effects of recombinant human growth hormone on cirrhotic rats.

AIM: To investigate the effects and molecular mechanisms of recombinant human growth hormone (rhGH) on protecting liver function and alleviating portal hypertension of liver cirrhotic rats. METHODS: Liver cirrhosis of male Sprague-Dawley rats was induced by administration of thioacetamide. The rats with or without liver cirrhosis were randomly divided into four groups. Group A consisted of the normal rats was treated with normal saline (NS), group B consisted of the normal rats was treated with rhGH, group C consisted of cirrhotic rats was treated with NS, and group D consisted of cirrhotic rats was treated with rhGH. The rats of different groups were subcutaneously injected with 0.5 mL of NS or 333 ng/kg of rhGH daily for 7 d. After treatments, the following parameters were examined, including GH-binding capacity (R(T)) by (125)I-hGH binding, growth hormone receptor mRNA(GHR mRNA) expression by RT-PCR, relative content of collagen (RCC) by histomorphomertry, and level of malon-dialdehyde (MDA) and superoxide dismutase (SOD) in liver tissue by thiobarbituric acid reaction and pyrogallic acid self-oxidation, respectively. Serum albumin (ALB), alanine transaminase (ALT) and portal vein pressure (PVP) were also examined. RESULTS: rhGH up-regulated both the GH-binding capacity (R(T)) and the expression of GHR mRNA in vivo. R(T) in group A (72+/-12 fmol/mg protein) was significantly higher than that in group C (31+/-4 fmol/mg protein) (P<0.05). R(T) in group B (80+/-9 fmol/mg protein) increased markedly compared to group A (P<0.05). R(T) in group D (40+/-7 fmol/mg protein) raised remarkably compared with group C (P<0.05), but less than that in group A, and there was no significant GH binding affinity contrast (Kd) change. The GHR mRNA level (iOD, pixel) in group A (29+/-3) was significantly higher than that in group C (23+/-3) (P<0.05). GHR mRNA levels were significantly raised in group B (56+/-4) and group D (42+/-8) compared with groups A and C (29+/-3 and 23+/-3, respectively) (P<0.05). Compared with the normal liver, MDA level was higher and SOD level was lower in cirrhotic livers. After rhGH treatment, MDA level was significantly declined to 12.0+/-2.2 nmol/mg protein and SOD was raised to 1 029+/-76 U/mg protein in group D (P<0.05). ALB levels in groups B and D (42+/-7 g/L and 37+/-7 g/L, respectively) were significantly raised compared with those in groups A and C (35+/-5 g/L and 29+/-4 g/L, respectively) (P<0.05). ALT level was markedly lower in group D (69+/-7 U/L) compared to group C (89+/-15 U/L) (P<0.05), and close to group A (61+/-10 U/L). RCC in group C (22.30+/-3.86%) was significantly higher than that in group A (1.14+/-0.21%) and group D (14.70+/-2.07%) (P<0.05). In addition, rhGH markedly alleviated portal hypertension in liver cirrhotic rats (group D vs C, 9.3+/-1.5 cmH(2)O vs 14.4+/-2.0 cmH(2)O) (P<0.05). CONCLUSION: Pharmacological doses of rhGH can increase R(T) and GHR mRNA expression, ameliorate liver functions, repress fibrosis and decline portal hypertension, suggesting it has potentially clinical usage as a hepatotropic factor.

Animals↗

Effects of recombinant human growth hormone on remnant liver after hepatectomy in hepatocellular carcinoma with cirrhosis.

AIM: To explore the effects of recombinant human growth hormone (rhGH) on the remnant liver after hepatectomy in hepatocellular carcinoma with liver cirrhosis. METHODS: Twenty-four patients with hepatocellular carcinoma who underwent hepatectomy were randomly divided into 2 groups: parenteral nutrition (PN) group (n=12) and rhGH+PN group (n=12). Liver function, blood glucose, AFP, serum prealbumin and transferrin were detected before operation, at post-operative d 1 and d 6. Albumin (ALB) mRNA in liver biopsy specimens was detected by RT-PCR at post-operative d 6. Liver Ki67 immunohistochemical staining was studied. RESULTS: On post-operative d 6, compared with PN group, the levels of blood glucose, serum prealbumin, transferrin, the expression of hepatic ALB mRNA and liver Ki67 labeling index were higher in rhGH+PN group. CONCLUSION: rhGH can improve protein synthesis and liver regeneration after hepatectomy in hepatocellular carcinoma with liver cirrhosis.

Adult↗

Protective effects of 5,4'-dihydroxy-3',5'-dimethoxy-7-O-beta-D -glucopyranosyloxy-flavone on experimental hepatic injury.

AIM: To investigate the pharmacological effects of rice flavone (5,4'-dihydroxy-3',5'-dimethoxy-7-O-beta-D-glucopyranosyloxy-flavone, RF) separated from panicle-differentiating to flowing rice on rat experimental hepatic injury. METHODS: Models of rat acute hepatic injury induced by carbon tetrachloride (CCl(4)) administration, rat hepatic fibrosis induced by thioacetamide, injury of primary cultured rat hepatocytes induced by CCl(4), respectively, were established. After treated with RF, content of serum alanine transaminase (ALT), aspartate aminotransferase (AST) and albumin (Alb), hyaluronic acid (HA), the activity of glutathione peroxidase (GSH-Px), superoxide dismutase (SOD), and hydroxyproline (Hyp) were measured and liver tissue was observed pathologically by hematoxylin-eosin (HE) staining. Effects of RF on pathological changes, function index, enzyme of scavenging free radicals and blood rheology were evaluated. RESULTS: In model of rat acute hepatic injury induced by CCl(4), RF can significantly decrease the contents of serum ALT, AST, increase the content of Alb, improve the dropsy and fat denaturalization of hepatocytes. In model of rat hepatic fibrosis induced by thioacetamide, RF can inhibit the increase of HA, Hyp and whole blood viscosity, and improve the activities of GSH-Px and SOD, and inauricular microcirculation. CONCLUSION: RF has apparent protective effects on hepatic injury by increasing activity of GSH-Px and SOD, scavenging free radicals produced by CCl(4), reducing blood viscosity, and improving microcirculation and blood supply.

Acute Disease↗

Multicenter clinical study on Fuzhenghuayu capsule against liver fibrosis due to chronic hepatitis B.

AIM: To study the efficacy and safety of Fuzhenghuayu capsule (FZHY capsule, a capsule for strengthening body resistance to remove blood stasis) against liver fibrosis due to chronic hepatitis B. METHODS: Multicenter, randomized, double blinded and parallel control experiment was conducted in patients (aged from 18 to 65 years) with liver fibrosis due to chronic hepatitis B. Hepatic histologic changes and HBV markers were examined at wk 0 and 24 during treatment. Serologic parameters (HA, LM, P-III-P, IV-C) were determined and B ultrasound examination of the spleen and liver was performed at wk 0, 12 and 24. Liver function (liver function and serologic parameters for liver fibrosis) was observed at wk 0, 6, 12, 18 and 24. Blood and urine routine test, renal function and ECG were examined before and after treatment. RESULTS: There was no significant difference between experimental group (110 cases) and control group (106 cases) in demographic features, vital signs, course of illness, history for drug anaphylaxis and previous therapy, liver function, serologic parameters for liver fibrosis, liver histologic examination (99 cases in experimental group, 96 cases in control group), HBV markers, and renal function. According to the criteria for liver fibrosis staging, mean score of fibrotic stage(s) in experimental group after treatment (1.80) decreased significantly compared to the previous treatment (2.33, P<0.05), but there was no significant difference in mean score of fibrotic stage(s) (2.11 and 2.14 respectively). There was a significant difference in reverse rate between experimental group (52%) and control group (23.3%) in liver biopsy. With marked effect on decreasing the mean value of inflammatory activity and score of inflammation (P<0.05), Fuzhenghuayu capsule had rather good effects on inhibiting inflammatory activity and was superior to that of Heluoshugan capsule. Compared to that of pretreatment, there was a significant decrease in HA, LM, P-III-P and IV-C content in experimental group after 12 and 24 wk of treatment. The difference in HA, LM, P-III-P and IV-C content between 12 and 24 wk of treatment and pretreatment in experimental group was significantly greater than that in control group (P<0.01-0.05). The effect, defined as two of four parameters lowering more than 30% of the baseline, was 72.7% in experimental group and 27.4% in control group (P<0.01). Obvious improvement in serum Alb, ALT, AST and GGT was seen in two groups. Compared to that of control group, marked improvement in GGT and Alb was seen in experimental group (P<0.05). The effective rate of improvement in serum ALT was 72.7% in experimental group and 59.4% in control group. No significant difference was seen in blood and urine routine and ECG before and after treatment. There was also no significant difference in stable rate in ALT and serologic parameters for liver fibrosis between experimental group and control group after 12 wk of withdrawal. CONCLUSION: Fuzhenghuayu capsule has good therapeutic effects on alleviating liver fibrosis due to chronic hepatitis B without any adverse effect and is superior to that of Heluoshugan capsule.

Adolescent↗

Porcine acute liver failure model established by two-phase surgery and treated with hollow fiber bioartificial liver support system.

AIM: To establish a highly reproducible animal model of acute liver failure (ALF), for assessing the effect of bioartificial liver support system (BALSS). METHODS: A two-phase complete liver devascularization procedure was performed in eight loco-hybrid pigs. Blood biochemical index and liver biopsy were studied every 2 h after surgery, and survival time was recorded. The BALSS constructed with high volume recirculating technique was a hollow fiber circulating system consisting of a hepatocyte reactor-hollow fiber module inoculated with microcarrier-adhering hepatocytes, and a double pump, heparinized, thermostabilized, micro-capsulized activated carbon-adsorbing plasmapheresis system. Twelve pigs undergoing two-phase surgery were randomized into: control group (perfused without hepatocytes, n = 6) and treatment group (perfused with hepatocytes, n = 6). Intergroup liver biochemical indexes, survival time, and liver pathological changes were analyzed at regular intervals. RESULTS: Two-phase surgery was performed in all the experimental pigs, and there was no obvious difference between their biochemical indexes. After 3 h of phase II surgery, ammonia (Amm) increased to (269+/-37) micromol/L. After 5 h of the surgery, fibrinogen (Fib) decreased to (1.5+/-0.2) g/L. After 7 h of the surgery, ALT, AST, Tbil and PT were (7.6+/-1.8) nka/L, (40+/-5) nka/L, (55+/-8) micromol/L and (17.5+/-1.7) nka/L respectively. After 9 h of surgery, ALB and Cr were (27+/-4) g/L and (87+/-9) micromol/L. After 13 h of surgery, BUN was (3.5+/-0.9) micromol/L. All the above values were different from those determined before surgery. Survival time of pigs averaged 13.5+/-1.4 h. ALF pigs in the other group were treated with BALSS. The comparison analysis between the treated and control animals showed the changes of Tbil, PT, Alb, BUN, Cr, Fib, and Amm (P<0.01), but there was no change of ALT and AST. The survival time was statistically different (P<0.01), and there was no significant difference in histological changes. CONCLUSION: The porcine ALF model established by two-phase devascularized surgery is valid and reproducible. The hollow fiber BALSS can meet the needs of life support and is effective in treating ALF.

Alanine Transaminase↗

Promoted differentiation of cynomolgus monkey ES cells into hepatocyte-like cells by co-culture with mouse fetal liver-derived cells.

AIM: To explore whether a co-culture of cynomolgus monkey embryonic stem (cES) cells with embryonic liver cells could promote their differentiation into hepatocytes. METHODS: Mouse fetal liver-derived cells (MFLCs) were prepared as adherent cells from mouse embryos on embryonic d (ED) 14, after which undifferentiated cES cells were co-cultured with MFLCs. The induction of cES cells along a hepatic lineage was examined in MFLC-assisted differentiation, spontaneous differentiation, and growth factors (GF) and chemicals-induced differentiations (GF-induced differentiation) using retinoic acid, leukemia inhibitory factor (LIF), FGF2, FGF4, hepatocyte growth factor (HGF), oncostatin M (OSM), and dexamethasone. RESULTS: The mRNA expression of alpha-fetoprotein, albumin (ALB), alpha-1-antitrypsin, and hepatocyte nuclear factor 4alpha was observed earlier in the differentiating cES cells co-cultured with MFLCs, as compared to cES cells undergoing spontaneous differentiation and those subjected to GF-induced differentiation. The expression of cytochrome P450 7a1, a possible marker for embryonic endoderm-derived mature hepatocytes, was only observed in cES cells that had differentiated in a co-culture with MFLCs. Further, the disappearance of Oct3/4, a representative marker of an undifferentiated state, was noted in cells co-cultured with MFLCs, but not in those undergoing spontaneous or GF-induced differentiation. Immunocytochemical analysis revealed an increased ratio of ALB-immunopositive cells among cES cells co-cultured with MFLCs, while glycogen storage and urea synthesis were also demonstrated. CONCLUSION: MFLCs showed an ability to induce cES cells to differentiate toward hepatocytes. The co-culture system with MFLCs is a useful method for induction of hepatocyte-like cells from undifferentiated cES cells.

Animals↗

Functional evaluation of a new bioartificial liver system in vitro and in vitro.

AIM: To evaluate the functions of a new bioartificial liver (BAL) system in vitro and in vitro. METHODS: The BAL system was configured by inoculating porcine hepatocyte spheroids into the cell circuit of a hollow fiber bioreactor. In the experiments of BAL in vitro, the levels of alanine aminotransferase (ALT), total bilirubin (TB), and albumin (ALB) in the circulating hepatocyte suspension and RPMI-1640 medium were determined during 6 h of circulation in the BAL device. In the experiments of BAL in vitro, acute liver failure (ALF) model in canine was induced by an end-side portocaval shunt combined with common bile duct ligation and transaction. Blood ALT, TB and ammonia levels of ALF in canines were determined before and after BAL treatment. RESULTS: During 6 h of circulation in vitro, there was no significant change of ALT, whereas the TB and ALB levels gradually increased with time both in the hepatocyte suspension and in RPMI-1640 medium. In the BAL treatment group, blood ALT, TB and ammonia levels of ALF in canines decreased significantly. CONCLUSION: The new BAL system has the ability to perform liver functions and can be used to treat ALF.

Alanine Transaminase↗

Protective effect of early enteral feeding on postburn impairment of liver function and its mechanism in rats.

AIM:To study the protective effect of early enteral feeding (EEF) on the postburn impairment of liver function and its mechanism.METHODS:Wistar rats with 30% of total body surface area (TBSA) full-thickness burn were employed. The effects of EEF on the postburn changes of gastric intramucosal pH, endotoxin levels in portal vein, water contents of hepatic tissue, blood concentrations of tumor necrosis factor (TNF-alpha), plasma activities of alanine aminotransferase (ALT) and asparate aminotransferase (AST), as well as the blood contents of total (TB) and direct bilirubin (DB), total protein (TP) and albumin (ALB) were serially determined within 48h postburn.RESULTS:EEF could significantly improve gastric mucosal acidosis, reduce portal vein endotoxin level and water content of hepatic tissue, as well as plasma concentrations of TNF-alpha at all timepoints after severe burns (P < 0.01); postburn elevation of the plasma activities of ALT, AST and the contents of TB, DB were effectively prevented, whereas the plasma concentrations of TP and ALB were markedly increased 24h and 48h posturn in EEF group compared with that of the burn without EEF group (P < 0.01 ).CONCLUSION:EEF has significant beneficial effects on the improvement of hepatic function in rats after severe burn, and is probably related with an increase in splanchnic blood flow, reduction of the absorption of gut-origin endotoxin and the consequent release of inflammatory mediators.

Journal Article↗

Generation and characterization of transgenic mice expressing tamoxifen-inducible cre-fusion protein specifically in mouse liver.

AIM: To establish transgenic mice expressing tamoxifen-inducible Cre-ERt recombinase specifically in the liver and to provide an efficient animal model for studying gene function in the liver and creating various mouse models mimicking human diseases. METHODS: Alb-Cre-ERt transgenic mice were produced by microinjecting the construct with Cre-ERt fusion gene of DNA fragments into fertilized eggs derived from inbred C57BL/6 strain. Transgenic mice were identified by using PCR and Southern blotting. Expression of Cre-ERt fusion gene was analyzed in the liver, kidney, brain and lung from F1 generation transgenic mice at 8 weeks of age by reverse transcription (RT)-PCR. RESULTS: Four hundred and fourteen fertilized eggs of C57 BL/6 mice were microinjected with recombinant Alb-Cre-ERt DNA fragments, and 312 survival eggs injected were transferred to the oviducts of 12 pseudopregnant recipient mice, 6 of 12 recipient mice became pregnant and gave birth to 44 offsprings. Of the 44 offsprings, two males and one female carried the hybrid Cre-ERt fusion gene. Three mice were determined as founders, and were back crossed to set up F1 generations with other inbred C57BL/6 mice. Transmission of Cre-ERt fusion gene in F1 offspring followed Mendelian rules. The expression of Cre-ERt mRNA was detected only in the liver of F1 offspring from two of three founder mice. CONCLUSION: Transgenic mice expressing tamoxifen-inducible Cre-ERt recombinase under control of the liver-specific promoter are preliminary established.

Animals↗

Does loss of residual renal function lead to malnutrition in peritoneal dialysis patients?

OBJECTIVES: It is usually believed that loss of residual renal function is associated with anorexia and the development of malnutrition. We conducted a retrospective study in our center to evaluate the effect of declining residual renal function on patients' nutritional status. METHODS: All incident uremic patients (n = 46) who began peritoneal dialysis from January 1, 2003 June 1, 2003 in our center were closely followed for 1 year with focus on maintaining strict volume control with time on dialysis. Patient's residual renal function (RRF) was assessed by the average renal urea and creatinine clearances. Those patients who had more than 50% decrease in GFR were selected for the present analysis. Serum albumin (ALB), dietary protein intake (DPI) and subjective global assessment (SGA) were closely followed. RESULTS: There were 16 patients (9 males and 7 females) included in the present analysis, among whom 31.3% were diabetics. Patients' GFR declined significantly (RRF were 4.32 +/- 2.69, 2.99 +/- 2.21 and 1.24 +/- 0.99 ml/min for Months 1, 6 and 12, respectively, p < 0.05), along with a significant decline in urine volume (985.62 +/- 543.29, 698.13 +/- 463.59 and 425.63 +/- 320.52 ml/d for Months 1, 6 and 12, respectively, p < 0.01). Although weekly peritoneal Kt/V did not increase significantly, peritoneal ultrafiltration increased significantly during this period (428.75 +/- 408.96, 534.38 +/- 296.39, 844.38 +/- 440.35 ml for Months 1, 6 and 12, respectively, p < 0.05). Serum ALB increased significantly (32.34 +/- 5.07, 34.74 +/- 4.89 and 36.21 +/- 3.98 g/l for Months 1, 6 and 12, respectively, p < 0.01). DPI also increased significantly. The prevalence of malnutrition (by SGA) decreased from 62.5% at the start of dialysis to 18.8% at the end of this study (p < 0.05). CONCLUSIONS: Our study suggests that rapid decline of residual renal function in PD patients does not necessarily lead to decreased dietary protein intake and deteriorated nutritional status. Focus on incremental peritoneal fluid removal along with the decline in residual renal function and, thus, maintaining volume control may be one of the critical reasons for the success.

Antihypertensive Agents↗

Changes in mallard (Anas platyrhynchos) serum chemistry due to age, sex, and reproductive condition.

Selected serum constituents were analyzed from 50 adult mallards (Anas platyrhynchos) of both sexes during several stages of reproduction: pre-egg laying, egg laying, incubating, molting, and postreproductive. Similar assays were conducted on sera from ducklings aged 5 to 58 days. Values for total protein (TPR), albumin (ALB), glucose (GLU), gamma-glutamyl transferase (GGT), calcium (CA), phosphorus (PHOS) and magnesium (MG) differed by sex. When all data were combined and analyzed for sex-related differences within each reproductive condition separately, all assays except lactate dehydrogenase (LD-L), cholinesterase (CHE), alanine aminotransferase (ALT), aspartate aminotransferase (AST), creatinine (CRN) and direct bilirubin (BIDI) differed between sexes during one or more reproductive periods. Each assay showed differences among the various reproductive conditions regardless of gender. The pattern of change differed between sexes. All assays except ALB, GLU, CA and MG showed age-related changes. Lipemia in the sample interfered with all chemistries except TPR, LD-L and CA. Results indicate that when using clinical chemistry as a diagnostic tool in the mallard, age and reproductive condition should be determined in order to compare the data to appropriate control values.

Age Factors↗

[Genetic linkage of blood group, egg and serum protein and plumage color loci in chickens].

Genetic relationship of six blood group (A, B, C, D, E, x5), three egg (G2, G3, Ov) and one serum (Alb) protein loci and two plumage colour (I-dominant white, E-extended black) loci were investigated. 3250 gametes have been analysed for 21 loci combinations, 11 from them have never been studied on linkage. Blood group loci A, B, C, D, E, x5 segregated independently on egg protein loci G2, G3, and Ov, serum protein locus Alb and plumage colour locus E. No linkage was observed between blood group locus B and dominant white locus I. Close linkage for two egg protein loci G3 and Ov is confirmed. Independent segregation of investigated blood group, egg and serum protein loci suggests their localization on different autosomes in the chicken genome. The recent literature and the authors' data on genetic relationship between blood group, polymorphic protein loci and morphological traits are reviewed.

Animals↗

Pharmacologically stimulated portal flow measurement by magnetic resonance imaging for assessment of liver function.

PURPOSE: To evaluate pharmacologically stimulated portal flow measured by magnetic resonance (MR) imaging for assessment of liver function. MATERIALS AND METHODS: Pharmacologically stimulated portal flow was measured by phase contrast MR imaging in 27 patients when they were undergoing abdominal angiography for liver tumors or gall bladder cancer. The patients included 11 cases of liver cirrhosis and eight of chronic hepatitis. Pharmacological stimulation was done by infusion of 10 microg/Kg of nicardipine hydrochloride into the superior mesenteric artery through an angiographic catheter. We examined the correlation between stimulated or non-stimulated portal flow and biochemical liver function tests. RESULTS: Correlation coefficients and their corresponding p values between non-stimulated portal flow and the indocyanine green residual rate at 15 min after injection (ICG R15), serum albumin (ALB), total bilirubin (TB), cholinesterase (CHE), and hepaplastin test (HP) were--0.414 (0.056), 0.296 (0.134), -0.570 (0.002), 0.289 (0.153), and 0.321 (0.126), respectively, whereas those between stimulated portal flow and ICG R15, ALB, TB, CHE, and HP were--0.561 (0.007), 0.411 (0.033), -0.509 (0.007), 0.445 (0.023), and 0.494 (0.014), respectively. CONCLUSION: Stimulated portal flow showed better correlations with biochemical liver function tests than non-stimulated portal flow. It is suggested that stimulated portal flow measurement is more useful for the evaluation of liver function than non-stimulated portal flow measurement.

Aged↗

[Evaluation of screening methods for cholinesterase variants from daily laboratory data].

The genetic variants of the cholinesterase (ChE) are frequently misdiagnosed as a liver dysfunction. We compared three extraction methods for screening of ChE abnormalities. Employing these three methods, total 31 cases were found to be genetic abnormalities from 2985 patients of Hamamatsu University Hospital. We picked up 11 of candidates with low enzyme activities less than 100U/l as group 1 using the first method and effectively detected 9 cases (82%) with genetic abnormalities. The second extraction method was based on the ratio between Albumin (Alb) and ChE and subsequently, 48 of high risk patients were picked up as group 2 and 28 cases (58%) showed genetic abnormalities. Furthermore, all cases of group 1 were contained in the second group. The third method was based on the discrimination function from Alb and total cholesterol (TC) as group 3 and 32 cases were picked up. Fourteen cases (44%) out of them showed the genetic abnormalities using this method, and surprisingly, 13 cases (93%) of them were estimated to be K-variant. Although the three methods showed the different characteristics to extract genetic abnormalities of ChE, the second extraction method could pick up the largest population with genetic abnormalities. Further phenotypic extraction methods should be compared to understand the relationship between phenotype and genotype.

Cholesterol↗

[Set-up of standard value by sex and age of biochemical tests derived from blood donors].

We set up the standard value of 7 biochemical tests from the data of approximately 237,000 blood donors who are healthy at least objectively. The abnormal data were excluded by Grubbs Smirnov method, and then converted by the log equation in order to simulate to normal distribution. Then the reference interval was set at mean +/- 2SD. If this failed, 95% percentile was adopted after non-parametric analysis. The data fluctuated much between sex and generations. Especially sex difference was dominant in ALT, AST, gamma-GTP and TP. ALT and AST was greater in males than in females, although age difference was smaller with aging. gamma-GTP peaked in 40s in males and greater in males than in females. TP was greater in males. Age difference was dominant in ALB, CHOL and BUN. ALB decreased with aging, and CHOL and BUN increased with aging. Therefore standard value in each sex and age group should be adopted in these tests. The volume of the data in each age group in this study is suitable for the set-up of the standard value.

Adolescent↗

[Transition of branched-chain amino acids and tyrosine ratio (BTR) in the blood of acute hepatitis patients].

The molar ratio of branched-chain amino acids to tyrosine (BTR) correlates well with the Fischer ratio, and can be measured in a short period of time. It is regarded as the method of analysis that will eventually replace the Fischer ratio. But clinical significance of BTR in terms of acute liver disorders has not been examined thoroughly as of yet. In this study, we measured BTR of 34 patients with acute hepatitis, and examined the transition of the acute period of acute hepatitis and its recovery process. Thirty-four patients diagnosed with acute viral hepatitis became subjects of examination (16 patients of A type, 15 patients of B type, 1 patient of C type, 2 patients of non-A, non-B, non-C type). Out of the 34 patients, 11 were in serious stages (HPT under 40%), including 3 in fulminant condition. By using preserved serum obtained during the acute period (within 1 week of the highest transaminase value), recovery period (within 4 weeks), and treatment period (3 months and later), measurements were conducted with Diacolor:BTR (enzymatic analysis, ONO Pharmaceutical Co., Ltd.), and the results were compared with those of 50 healthy subjects (25 men, 25 women). BTR correlated well with the Fischer ratio for chronic hepatic patients, and with albumin (Alb), PT, and ICGR15 as well, proving that it is useful as an indicator of hepatic reserve ability. But BTR has not been thoroughly examined as it relates to acute liver disorders. In this study, BTR fell in the acute period, correlating with the serious period, proving that it is a useful indicator. For acute liver damage, BTR supports conventional indicators (Alb, Ch-E, HGF, etc.) for assessing serious damage. Also, it has been suggested that measuring the passage of BTR could be the indicator of true recovery, including amino acid metabolism for liver disorders.

Acute Disease↗

[Effects of plasma endothelin-1 and aldosterone on sodium retention in children with nephrotic syndrome].

The effects of plasma endothelin-1 (ET-1) and aldosterone (Ald) on sodium retention in children with nephrotic syndrome were investigated. 14 cases were divided into the edematous stage and non-edematous stage. Plasma ET-1, Ald, and serum osmolarity, albumin (Alb), and Na+ were measured in each stage. The results showed that the plasma ET-1 in edemaous stage increased significantly, compared with that in no-edematous stage (P < 0.01). There was no significant difference in plasma Ald between the edematous stage and the non-edematous stage. Plasma ET-1 was positively correlated with the serum sodium ionic concentration and negatively correlated with the serum Alb(r = 0.486, P < 0.01; r = 0.490, P < 0.01, respectively). In conclusion, the sodium retention with nephrotic syndrome might be correlated with reduction of secreted sodium in the kidney, suggesting that ET-1 plays an important role in pathogenesis.

Aldosterone↗