N-Terminal PreS1 Sequence Regulates Efficient Infection of Cell-Culture-Generated Hepatitis B Virus.

BACKGROUND AND AIMS: An efficient cell-culture system for hepatitis B virus (HBV) is indispensable for research on viral characteristics and antiviral reagents. Currently, for the HBV infection assay in cell culture, viruses derived from HBV genome-integrated cell lines of HepG2.2.15 or HepAD-38 are commonly used. However, these viruses are not suitable for the evaluation of polymorphism-dependent viral characteristics or resistant mutations against antiviral reagents. HBV obtained by the transient transfection of the ordinary HBV molecular clone has limited infection efficiencies in cell culture. APPROACH AND RESULTS: We found that an 11-amino-acid deletion (d11) in the preS1 region enhances the infectivity of cell-culture-generated HBV (HBVcc) to sodium taurocholate cotransporting polypeptide-transduced HepG2 (HepG2/NTCP) cells. Infection of HBVcc derived from a d11-introduced genotype C strain (GTC-d11) was ~10-fold more efficient than infection of wild-type GTC (GTC-wt), and the number of infected cells was comparable between GTC-d11- and HepG2.2.15-derived viruses when inoculated with the same genome equivalents. A time-dependent increase in pregenomic RNA and efficient synthesis of covalently closed circular DNA were detected after infection with the GTC-d11 virus. The involvement of d11 in the HBV large surface protein in the enhanced infectivity was confirmed by an HBV reporter virus and hepatitis D virus infection system. The binding step of the GTC-d11 virus onto the cell surface was responsible for this efficient infection. CONCLUSIONS: This system provides a powerful tool for studying the infection and propagation of HBV in cell culture and also for developing the antiviral strategy against HBV infection.

Effects of traditional Chinese medicine formula Le-Cao-Shi on hepatitis B: In vivo and in vitro studies.

ETHNOPHARMACOLOGICAL RELEVANCE: Formula Le-Cao-Shi (LCS) is a traditional Chinese medicine (TCM), which has long been used as a folk remedy against hepatitis B in China. The present study was conducted to evaluate the anti-hepatitis B effects of aqueous extract of LCS in vivo and in vitro. MATERIALS AND METHOD: we investigated the anti-HBV effects of LCS in vivo and in vitro with duck hepatitis B model and HepG2.2.15 cell line model, respectively. The serologic and cellular biomarkers and the histopathological changes were examined. RESULTS: By a duck hepatitis B model, the extract of LCS was found to restrain the expressions of duck hepatitis B surface antigen (DHBsAg), hepatitis B e antigen (DHBeAg), and HBV-DNA (DHBV-DNA). Moreover, LCS could decrease the levels of aspartate and alanine aminotransferases (AST and ALT) and ameliorate duck liver histological lesions. Correspondingly, in a HepG2.2.15 cellular model, LCS could also significantly inhibit the secretions of HBsAg and HBeAg. CONCLUSION: LCS exerted potent anti-hepatitis effects against the infection of HBV. The above results demonstrated the first-hand experimental evidences for the anti-hepatitis B efficiency of LCS. Our study provides a basis for further exploration and development of this promising compound prescription to treat hepatitis B disease.

Hepatoprotective and inhibiting HBV effects of polysaccharides from roots of Sophora flavescens.

Roots of Sophora flavescens is an important herbal medicine for treatment of HBV and hepatic carcinoma in China. Alkaloids in the root were well known for exhibiting good hepato-protective and anti-HBV effects. However, polysaccharides as main components in the root remained unknown. In the studies, we investigated the chemical features and hepatoprotective effects of Sophora flavescens polysaccharides (SFP-100 and its active fractions) with ConA-induced hepatitis mice, human liver LO2 cells and HepG2.2.15 cells. The results showed that SFP-100 was composed of arabinose, glucose, galactose and galacturonic acid, SFP-100-A mainly contained glucose. SFP-100-B and SFP-100-C were acidic polysaccharides. SFP-100 significantly decreased hepatocytes apoptosis, inhibited the infiltration of neutrophils and macrophages into liver, and improved the production of IFN-γ and IL-6 of splenocytes in ConA-induced hepatitis mice. SFP-100 and its two sugar fractions increased LO2 cell proliferation and reduced cell apoptosis induced by ConA. SFP-100, SFP-100-A and SFP-100-C remarkedly inhibited the secretion of HBsAg and HBeAg by HepG2.2.15 cells.These results suggested Sophora flavescens polysaccharides exerts significant hepatoprotective and anti-HBV roles, and further is used for treatment of immune-mediated liver disease in the future.

Study of ZHENG differentiation in hepatitis B-caused cirrhosis: a transcriptional profiling analysis.

BACKGROUND: In traditional Chinese medicine (TCM) clinical practice, ZHENG (also known as TCM syndrome) helps to understand the human homeostasis and guide individualized treatment. However, the scientific basis of ZHENG remains unclear due to limitations of current reductionist approaches. METHODS: We collected the leukocyte samples of three hepatitis B-caused cirrhosis (HBC) patients with dampness-heat accumulation syndrome (DHAS) and three HBC patients with liver depression and spleen deficiency syndrome (LDSDS) for microarray analysis. We generated Gene-Regulatory-Networks (GeneRelNet) from the differentially expressed genes (DEGs) of microarray date. Core genes were validated using anther independent cohort of 40 HBC patients (20 DHAS, 20 LDSDS) with RT-PCR. RESULTS: There were 2457 mapped genes were differentially expressed between DHAS and LDSDS (Fold change ≥ 2.0, P < 0.05). There were markedly different genes co-expression patterns in DHAS and LDSDS. Furthermore, three differential co-expression genes including purine nucleoside phosphorylase (PNP); aquaporin 7 (AQP7) and proteasome 26S subunit, non-ATPase 2 (PSMD2) were screened by GeneRelNets, and their mRNA expressions were further validated by real time RT-PCR. The results were consistent with microarray. The PNP (P = 0.007), AQP7 (P = 0.038) and PSMD2 (P = 0.009) mRNA expression is significant difference between DHAS and LDSDS using the non-parametric test. Furthermore, we constructed an mRNA panel of PNP, AQP7 and PSMD2 (PAP panel) by logistic regression model, and evaluated the PAP panel to distinguish DHAS from LDSDS by area under the receiver operating characteristic curve (AUC) analysis, which showed a higher accuracy (AUC = 0.835). Gene ontology (GO) analysis indicated that the DHAS is most likely related to system process while the functions overrepresented by LDSDS most related to the response to stimulus. CONCLUSIONS: This study suggested that there are particular transcriptional profiles, genes co-expressions patterns and functional properties of DHAS and LDSDS, and PNP, AQP7, and PSMD2 may be involved in ZHENG differentiation of DHAS and LDSDS in HBC.

Toll-like receptor 8 agonist and bacteria trigger potent activation of innate immune cells in human liver.

The ability of innate immune cells to sense and respond to impending danger varies by anatomical location. The liver is considered tolerogenic but is still capable of mounting a successful immune response to clear various infections. To understand whether hepatic immune cells tune their response to different infectious challenges, we probed mononuclear cells purified from human healthy and diseased livers with distinct pathogen-associated molecules. We discovered that only the TLR8 agonist ssRNA40 selectively activated liver-resident innate immune cells to produce substantial quantities of IFN-γ. We identified CD161(Bright) mucosal-associated invariant T (MAIT) and CD56(Bright) NK cells as the responding liver-resident innate immune cells. Their activation was not directly induced by the TLR8 agonist but was dependent on IL-12 and IL-18 production by ssRNA40-activated intrahepatic monocytes. Importantly, the ssRNA40-induced cytokine-dependent activation of MAIT cells mirrored responses induced by bacteria, i.e., generating a selective production of high levels of IFN-γ, without the concomitant production of TNF-α or IL-17A. The intrahepatic IFN-γ production could be detected not only in healthy livers, but also in HBV- or HCV-infected livers. In conclusion, the human liver harbors a network of immune cells able to modulate their immunological responses to different pathogen-associated molecules. Their ability to generate a strong production of IFN-γ upon stimulation with TLR8 agonist opens new therapeutic opportunities for the treatment of diverse liver pathologies.

Complementary laboratory indices for predicting the disease status of patients with hepatitis B virus infection.

To identify complementary laboratory indices for determining the disease status of patients with hepatitis B virus. Subjects were divided into six groups: hepatitis B virus carrier, mild chronic hepatitis B, moderate chronic hepatitis B, severe chronic hepatitis B, fulminant hepatitis B and healthy controls. Serum alanine aminotransferase, total bilirubin and direct bilirubin were measured by an automatic analyser. The levels of T-cell immunoglobulin domain and mucin-domain-containing molecule-3, macrophage inflammatory protein 2, neutrophil gelatinase-associated lipocalin and inducible nitric oxide synthase were measured by ELISA. T-cell immunoglobulin domain, mucin-domain-containing molecule-3, macrophage inflammatory protein 2 and inducible nitric oxide synthase levels were significantly higher in patients with severe chronic hepatitis B compared with those in patients with mild and moderate chronic hepatitis B or fulminant hepatitis B (P < 0.05). When normal or abnormal alanine aminotransferase was present, significant differences between macrophage inflammatory protein 2 and T-cell immunoglobulin domain and mucin-domain-containing molecule-3 levels between patients with mild, moderate, severe chronic hepatitis B or fulminant hepatitis B were observed (P < 0.05). Our results suggest that T-cell immunoglobulin domain and mucin-domain-containing molecule-3 and macrophage inflammatory protein 2 could serve as alanine aminotransferase, direct bilirubin or total bilirubin complementary indices for determining the status of patients with hepatitis B.

[Retrospective analysis of Mr. Xie Xiliang's medical records accumulated in 30 years on direct moxibustion for treating hepatitis B].

To analyze retrospectively Mr. Xie Xiliang's medical records accumulated in 30 years on direct moxibustion for treating hepatitis B, in which 86 cases were included. Before accepting direct moxibustion, 80.23% of the cases had been treated by conventional Chinese and western medicine, but no satisfactory therapeutic effects were obtained. Direct moxibustion was applied for 7-9 cones at each point selected. Point selection was related to ages. Usually, only a few points were used, and Ganshu (BL 18) and Pishu (BL 20) were used as main points. Shenzhu (GV 12) was added for children, and Zusanli (ST 36) added for adults. The improvement rate in clinical symptoms and physical signs were 100%. In the cases with hepatauxe, splenauxe, liver cirrhosis, and abdominal dropsy diagnozed by the B-ultrasonic examination, the improvement was obvious. Of the total, the rates turning to be negative in HBsAg, HBeAg and HBcAb were 28.85%, 38.46%, and 36.54%, respectively. It took some 10 days to 1 month to begin to show improvement in physical signs and hepetic functions, and about 3-6 months for their restoration. For the five items tested in type B hepatitis, it took 5 months to 1 year, even several years, to show their improvement. It is indicated that direct moxibustion is definitely effective in treating type B hepatitis. It is easy to handle and worth popularizing.

An inhibition of the hypothalamic somatostatinergic tone is not the cause of the enhanced growth hormone response to growth hormone-releasing hormone in patients with liver cirrhosis.

Animal models of liver cirrhosis (LC) display a reduced hypothalamic somatostatinergic tone. To test whether a similar mechanism could explain the enhanced Growth Hormone (GH) secretory response to GH-Releasing Hormone (GHRH), which is seen in human LC, we studied the effect of the cholinesterase inhibitor pyridostigmine (PD), which is able to reduce the release of hypothalamic somatostatin (SS), on the GHRH-stimulated GH secretion. We considered that if PD were unable to increase GH secretion, this would constitute evidence of an already inhibited endogenous somatostatinergic tone. If proved, this in turn could explain the enhanced GH response to GHRH seen in LC. Ten LC patients and nine controls were given GHRH (100 microg, intravenously), or PD (120 mg, orally) plus GHRH. After GHRH alone, the GH peak was four times higher in LC than in controls (40.85+/-15.7 ng/ml in LC and 9.35+/-2.5 ng/ml in controls). In LC, PD administration markedly increased the GH response to GHRH (GH peak: 98.0+/-19.7 ng/ml; +240% vs. GHRH alone). The ability of PD to increase the GH response in patients with LC suggests that in this condition the enhanced GH response to GHRH is not due to a completely inhibited endogenous somatostatinergic tone. SS appears instead to maintain its modulator role on GH secretion in human LC, in contrast with what observed in animal models.

Indications for biotherapy in systemic vasculitides.

Biotherapy now holds a specific place in the therapeutic armamentarium for systemic vasculitides. Such therapy includes cytokines, such as (pegylated) alpha-interferon for hepatitis B virus-related polyarteritis nodosa and hepatitis C virus-related cryoglobulinemic vasculitis, and polyvalent immunoglobulin (IVIg), with well-defined indications and pending positive results. More specifically targeted monoclonal antibodies include antitumor necrosis factor-alpha or anti-CD20 for antineutrophil cytoplasmic antibody-associated vasculitides or anti-interleukin-5 and anti-IgE for Churg-Strauss syndrome. However, the exact indications of these latter new agents, as well as their optimal dosage and duration, are not defined. Therefore, they are prescribed mainly for patients with disease refractory to conventional therapy, in whom results are promising. Results of international ongoing trials will determine whether the agents may also have a place as first-line treatment.

Hypoparathyroidism in transfusion-dependent patients with beta-thalassemia.

PURPOSE: To determine the prevalence of hypoparathyroidism in transfusion-dependent patients with beta-thalassemia. PATIENTS AND METHODS: A total of 28 transfusion-dependent patients with beta-thalassemia were interviewed, and their serum calcium, phosphate, magnesium, and intact parathyroid hormone levels were checked. Serum ferritin levels were measured to monitor the effect of chelation therapy. Blood urea nitrogen, creatinine, total protein, and albumin were measured in patients with undetectable or low intact parathyroid hormone levels. RESULTS: The prevalence of hypoparathyroidism was 10.7% (3/28). Mean age at diagnosis was 18 years. The serum ferritin levels of patients with hypoparathyroidism were 1,032, 2,102, and 7,680 microg/L. Only one patient had clinical symptoms of hypocalcemia. All three of the patients with hypoparathyroidism had hypogonadism, and 66.7% (2/3) of the patients had insulin-dependent diabetes mellitus. CONCLUSIONS: Hypoparathyroidism in transfusion-dependent patients with beta-thalassemia seems to be accompanied by other endocrinopathies. Serum ferritin may not have been a reliable indicator of iron overload in the three patients with hypoparathyroidism. Severe iron overload would easily explain these multiple endocrinopathies. This pattern is commonly seen in iron-overloaded patients with thalassemia elsewhere.

[Experimental study of inhibitory effect of kangxian recipe on TGF-beta 1 induced hepatocyte apoptosis].

OBJECTIVE: To study the inhibitory effect of Kangxian Recipe (KXR) on TGF-beta 1 induced hepatocyte apoptosis. METHODS: The in vitro model of hepatocyte apoptosis was established by cell biologic methods, utilizing the characteristics of TGF-beta 1 to observe the inhibitory effect of KXR on hepatocyte apoptosis. RESULTS: TGF-beta 1 induced apoptosis of hepatocyte in a dose-dependent manner. The apoptosis rate of 2.2.15 cells was 63% when 500 ng/L TGF-beta 1 was used, while for HepG2 cell, it was merely 44%. After treatment of 20 ng/L KXR, the apoptosis rate of the two kinds of cell lines lowered to 33% and 24% respectively. The inhibition rate of both groups was about 50%. CONCLUSION: KXR had strong inhibitory effect on hepatocyte apoptosis induced by TGF-beta 1.

Eficacia comparativa de las vías intradérmica e intramuscular en la inmunización activa contra la hepatitis B: resultados a largo plazo / Comparative efficacy of intradermally and intramuscular routes for active immunization against hepatitis B: long-term results

Se presentan los resultados a largo plazo de un estudio prospectivo, comparativo y randomizado con 119 voluntarios jóvenes sero-negativos, con riesgo aumentado de infección por el VHB, que evaluó la inmunogenicidad comparativa de una vacuna recombinante contra la hepatitis B, administrada por vía intramuscular (IM) o intradérmica (ID). La vacuna fue administrada en tres dosis consecutivas, en un esquema de inmunización de corta duración de 0,1 y 2 meses. Una dosis de refuerzo fue administrada al cabo de 1 año, por la misma vía utilizada en la inmunización original. La distribución por edad y sexo en ambos grupos fue similar. Los resultados obtenidos al cabo de las tres dosis mostraron tasas de seroconversión y seroprotección similares para ambos grupos (96 por ciento y 96 por ciento para la vía IM Vs. 98 por ciento y 93 por ciento para la vía ID, respectivamente). El promedio geométrico del título de anti-HBsAg en el suero de los individuos inmunizados por vía IM (155 UI/L) fue significativamente superior al obtenido en aquellos inmunizados por vía ID (71 UI/L). Por otro lado, la tasa de buenos respondedores (niveles de anti-HBsAg >100 UI/L ) fue igualmente más elevada en los vacunados por vía IM (67 por ciento Vs. 39 por ciento). El porcentaje de buenos respondedores en las mujeres fue consistentemente superior al de los varones para ambos grupos (75 por ciento Vs. 55 por ciento vía IM y 54 por ciento Vs. 13 por ciento vía ID, respectivamente). En 82 individuos estudiados 1 años más tarde, el promedio geométrico de los niveles de anti-HBsAg fue significativamente superior en los que recibieron la vía IM (278 UI/L) en comparación con los vacunados por vía ID (82,3 UI/L) y la tasa de seroprotección fue también superior (100 por ciento Vs. 84,8 por ciento respectivamente). De la misma manera, la respuesta de anti-HBsAg sérica a la dosis de refuerzo, fue mejor en los vacunados por vía IM (promedio geométrico 5.090 UI/L) con respecto a los que recibieron la vía ID.

Magnetization transfer contrast of various regions of the brain in liver cirrhosis.

BACKGROUND AND PURPOSE: T1-weighted MR images show high signal intensity in the pallidum of many patients with liver cirrhosis. The purpose of this study was to evaluate quantitative changes in MR signals in patients with liver cirrhosis by using the magnetization transfer technique. METHODS: Magnetization transfer ratios were measured in seven different regions of the brain in 37 patients with liver cirrhosis and in 37 healthy volunteers. RESULTS: The magnetization transfer ratios in patients with liver cirrhosis were significantly lower than those in control subjects in the globus pallidus, putamen, thalamus, corona radiata, and subcortical white matter. CONCLUSION: Abnormal magnetization transfer ratios may be found in otherwise normal-appearing cerebral regions.

Essential fatty acid supplementation in chronic hepatitis B.

BACKGROUND: Dietary supplementation with essential fatty acids and polyunsaturated lecithin may improve biochemical and histological parameters in liver disease. METHODS: Ten patients with serological and histological evidence of chronic hepatitis B received capsules of the polyunsaturated fatty acid-rich evening primrose oil in a dose of 4 g daily for 12 months, while a matched group received liquid paraffin capsules as a placebo. RESULTS: Compared to the placebo group, the patients receiving evening primrose oil showed no improvement in either biochemical or histological indices of liver damage, or in the rate of loss of circulating e antigen. CONCLUSIONS: Dietary, supplementation with this dose of essential fatty acids is unlikely to be of benefit in chronic hepatitis B.

Exacerbation of hepatitis in hepatitis B carriers following chemotherapy for haematological malignancies.

We describe nine asymptomatic chronic carriers of hepatitis B virus, four males and five females, with a mean age of forty-six years and all were Chinese, who developed exacerbation of hepatitis following chemotherapy for haematological malignancies. Seven patients had non-Hodgkin's lymphoma of whom three were treated with MACOP-B, two with BCEPP, one with PROMACE-CYTABOM and one with CHOP. Two patients had acute myeloid leukaemia and were treated with daunorubicin and cytosine arabinoside. Exacerbation of hepatitis occurred between one to four weeks following the last course of chemotherapy in eight patients. Two patients developed exacerbation of hepatitis when the dosage of prednisolone was reduced after they had ten weeks of high dose prednisolone. The outcome was fatal in six patients; all of whom developed hepatic encephalopathy. In four of these patients, alanine transaminase levels exceeded 1000 iu/l. Cytotoxic and immunosuppressive therapy permit enhanced viral replication. Withdrawal of the drugs results in partial restoration of immunocompetence and leads to rapid destruction of hepatocytes with consequent hepatic necrosis. Hence, patients who are hepatitis B virus carriers undergoing chemotherapy should be closely monitored. The fatal outcome of reactivation of chronic hepatitis B virus warrants prospective trials addressing preventive measures.

[Comparison of pre- and post-treatmental hepatohistology with heavy dosage of Paeonia rubra on chronic active hepatitis caused liver fibrosis].

No report has been found on comparison of pre- and post-treatmental liver biopsies for chronic active hepatitis (CAH) caused liver fibrosis. The aim of this paper was to study the effects of a heavy dosage of Radix Paeoniae rubra to the reabsorptive action of liver collagen fibres. The patients were suffering from CAH or CAH with liver cirrhosis confirmed by liver biopsies. By the end of 3 months, the second liver biopsy was carried out. The results showed that among 4 patients with liver cirrhosis, the false lobules disappeared; and that 6 patients with CAH only, the interlobular collagen fibres of 4 patients were completely reabsorbed. The effective rate reached 77.8%. It was concluded that a heavy dosage of Paeonia rubra was effective in arresting the development of liver fibrosis, and in promoting the reabsorption of collagen fibres.

[Relation between traditional Chinese medicine differentiation of syndromes and HBV antigen, HBV DNA in serum and liver tissues and pathological changes in chronic hepatitis B].

TCM differentiation of syndrome in 131 cases of chronic hepatitis B has been studied with molecular-biological and immuno-histological techniques. The results showed that 94.6% cases of Gan-Yu Pi-Xu (stagnancy in the liver leading to diminished function of the spleen) type belonged to chronic persistent hepatitis (CPH), whose coincidence rate of pathology of the liver biopsies with CPH was 69.2%; the positive rate of HBeAg and/or HBV DNA in sera was 61.5%, and the positive rates of HBsAg and HBV DNA in liver tissues were 69.2% (of which 44.4% appeared diffuse pattern morphologically) and 33.3% respectively. 75.5% cases of Gan-Shen Yin-Xu (deficiency of Yin of the liver and kidney) type belonged to chronic active hepatitis (CAH) and 88.5% of the cases were pathologically described as CAH, the positive rates of HBeAg and/or HBV DNA in serum and HBsAg in liver tissues were all 80.8%, among which the diffuse pattern of HBsAg accounted for 85.7%, which was higher than that in Gan-Yu Pi-Xu type (P less than 0.05), the positive rates of HBcAg and HBV DNA in liver tissues were 34.6% (of which 55.6% appeared cytoplasmic pattern) and 63.2% respectively, which was higher than that in Gan-Yu Pi-Xu type (P less than 0.05). 75.0% cases of Qi-Zhi Xue-Yu (stagnation of vital energy and stasis of blood) type belonged to CAH with early state cirrhosis, its pathological changes in liver tissues were obvious, replication levels of HBV corresponded to the cases of Gan-Yu.Pi-Xu type.

Effect of sho-saiko-to(xiao-chai-hu-tang) on HBeAg clearance in children with chronic hepatitis B virus infection and with sustained liver disease.

We studied the effect of Sho-saiko-to (Xiao-Chai-Hu-Tang) on HBeAg clearance rate (SN rate) in fourteen children with chronic hepatitis B virus (HBV) infection and with sustained liver disease. Seven of fourteen patients (50.0%) became HBeAg negative in the average observation period of 0.47 years(0.2-0.9 years). Four of those seven patients developed anti-HBe. The annual SN rate in the She-saiko-to treated group was apparently higher than the natural annual SN rate (22.7%) of 22 untreated patients retrospectively reviewed from the onset of hepatitis. Sho-saiko-to seemed to promote clearance of HBeAg in children with chronic HBV infection and with sustained liver disease. Sho-saiko-to may be a useful drug for such patients.

[Relation of changes in plasma cAMP, cGMP and the clinical conditions, pathology and the type of traditional Chinese medicine in 50 cases of chronic severe icteric hepatitis].

Plasma of cAMP and cGMP in 30 cases of chronic active hepatitis (group of cholestatic patients) as well as in 20 cases of chronic hepatitis with submassive or massive necrosis (group of chronic severe hepatitis patients) were studied with 125I-marked radioimmunoassay. 50 cases with their serum bilirubin levels higher than 171 mumol/L were selected as subjects for this study. Among them, 42 were diagnosed as hepatitis B, 6 as coinfection of hepatitis A and B. According to differentiation of symptoms and signs of TCM, 24 were diagnosed as simple hepatitis due to blood stasis and blood heat, 17 as hepatitis due to blood stasis and blood heat accompanying symptoms of Yang deficiency of the Spleen and Kidney. 20 healthy persons were selected as controls. The results were as follows: cAMP was 56.82 +/- 25.54 ng/L and 80.32 +/- 20.73 ng/L, and cGMP was 9.07 +/- 6.56 ng/L and 19.49 +/- 9.34 ng/L in the group of cholestatic patients and in the group of chronic severe hepatitis patients respectively. Both were higher than those in the control group whose cAMP was 14.12 +/- 3.25 ng/L and cGMP was 5.87 +/- 1.44 ng/L (P less than 0.01). Increase in cGMP and decrease in the ratio of cAMP and cGMP in the cases accompanying symptoms of Yang deficiency of the Spleen and Kidney were much higher than those in the other two types of hepatitis patients (P less than 0.01 and 0.05).(ABSTRACT TRUNCATED AT 250 WORDS)