Two new quinoline alkaloid with neuroprotective activities from Xylaria longipes.

Two new quinoline alkaloids with an α, ß-unsaturated amide side chain, xylarinines A and B (1 and 2), were isolated from the ethyl acetate extracts of Xylaria longipes solid fermentation. The structures of these were primarily determined though NMR and HRESIMS data analysis. The absolute configuration of compound 1 was assigned using experimental and calculated ECD data. The neuroprotective effects of compounds 1 and 2 against glutamate-induced damage in PC12 cells were evaluated in vitro bioassay. The results demonstrated that both compounds significantly improved cell viability, inhibited apoptosis, decreased malondialdehyde (MDA) levels, increased superoxide dismutase (SOD) and glutathione (GSH) levels, and reduced intracellular reactive oxygen species (ROS) accumulation. These findings suggested that these mechanisms contribute to the neuroprotective effects of the compounds.

The ethnopharmacology, phytochemistry and pharmacology of the genus Hericium.

ETHNOPHARMACOLOGICAL RELEVANCE: Mushrooms in the genus Hericium are used as functional food and traditional medicines for a long history in East Asian countries such as China, India, Japan, and Korea. Some species of Hericium are called as monkey head mushroom (Houtougu) in China and Yamabushitake in Japan, which are traditionally considered as rare and precious health promoting food and medicinal materials for the treatment of dyspepsia, insomnia, chronic gastritis, and digestive tract tumors. THE AIM OF THE REVIEW: This review aims to summarize the ethnopharmacology and structural diversity of secondary metabolites from Hericium species, as well as the pharmacological activities of the crude extracts and pure compounds from Hericium species in recent years. MATERIALS AND METHODS: All the information was gathered by searching Scifinder, PubMed, Web of Science, ScienceDirect, Springer, Wiley, ACS, CNKI, Baidu Scholar, Google Scholar databases and other published materials (books and Ph.D. and M. Sc. Dissertations) using the keywords "Hericium", "Traditional uses", "Chemical composition", "Quality control" and "Pharmacological activity" (1971-May 2023). The species name was checked with https://www.mycobank.org/. RESULTS: The traditional uses of Hericium species were summarized, and 230 secondary metabolites from Hericium species were summarized and classified into six classes, mainly focusing on their chemical diversity, biosynthesis, biological activities. The modern pharmacological experiments in vivo or in vitro on their crude and fractionated extracts showed that the chemical components from Hericium species have a broad range of bioactivities, including neuroprotective, antimicrobial, anticancer, α-glucosidase inhibitory, antioxidant, and anti-inflammatory activities. CONCLUSIONS: The secondary metabolites discovered from Hericium species are highly structurally diverse, and they have the potential to be rich resources of bioactive fungal natural products. Moreover, the unveiled bioactivities of their crude extracts and pure compounds are closely related to critical human health concerns, and in-depth studies on the potential lead compounds, mechanism of pharmacological effects and pharmaceutical properties are clearly warranted.

Liver-cell protective pyridones from the fungi Tolypocladium album dws120.

Five previously undescribed pyridone derivatives, tolypyridones I-M, were identified from the solid rice medium fermented by Tolypocladium album dws120, along with two known compounds tolypyridone A (or trichodin A) and pyridoxatin. Their planar structures and partial relative configurations have been determined by careful interpretation of their spectroscopic data. The full assignment of the relative and absolute configurations of tolypyridones I-M was achieved by gauge-independent atomic orbital 13C NMR calculation, quantitative nuclear Overhauser effects based interatomic distance calculation, and electronic circular dichroism calculation. In addition, we have fully determined the configuration of tolypyridone A by X-ray diffraction analysis. In bioassay, tolypyridones I was able to restore cell viability and inhibit the release of alanine aminotransferase and aspartate aminotransferase for ethanol-induced LO2 cells, suggesting its potential as a liver protective agent.

Two pairs of new isobenzofuranone enantiomers from a soil-derived fungus Penicillium canescens DWS225.

Two pairs of new isobenzofuranone derivative enantiomers, (±)-penicifurans E (1) and (±)-penicifurans F (2), together with four know compounds (3-6) were isolated from the solid fermentation of Penicillium canescens DWS225. The structures of these enantiomers were elucidated by extensive NMR spectroscopic data, and their absolute configurations were assigned by the experimental and calculated ECD data. The neuroprotective effects of all the isolates against oxygen-glucose deprivation/reperfusion injury in pheochromocytoma-12 cells (PC12) were investigated.

Neuroprotective methylsuccinic acid and enoic acid derivatives from the fungus Xylaria longipes.

Three undescribed methylsuccinic acid derivatives, xylaril acids A-C, and two undescribed enoic acid derivatives, xylaril acids D-E, were isolated from the fungus Xylaria longipes. The structures of the undescribed compounds were deduced by spectroscopic means, including HRESIMS and 1D/2D NMR spectroscopy, as well as ECD calculations. The absolute configuration of xylaril acids A was further determined by single-crystal X-ray diffraction experiments. All the isolated compounds displayed neuroprotective activities against oxygen-glucose deprivation/reperfusion injury in PC12 cells by enhancing cell viability and inhibiting cell apoptosis.

Reassignment of the structures of pestalopyrones A-D.

Pestalopyrones A-D are four unusual tricyclic pyrone derivatives with flexible chiral structures, isolated from the endophytic fungus Pestalotiopsis neglecta S3. The full elucidation of their structures was a challenging task, and remained unsolved in the original article. Herein, the relative configurations of pestalopyrones A and pestalopyrones B were unambiguously assigned by detailed analyses on spectroscopic data and GIAO 13C NMR calculation method with sorted training sets (STS). The planar structures of pestalopyrones C and pestalopyrones D were revised by reinterpretation of their reported spectroscopic data, and then their relative configurations were deduced by STS GIAO 13C NMR calculation and NOE analysis. The absolute configurations of all the mentioned compounds were determined by the comparison of their experimental and calculated ECD curves.

[The analysis of clinical character in different age patients suffered from A-H1N1].

OBJECTIVE: To describe the feature of different age patients with A-H1N1. METHODS: Cross-sectional study was performed in 95 patients who were confirmed to be infected with A-H1N1 from May, 2009 to July, 2009, in according to their age. RESULTS: The average age of patients with A-H1N1 infection was 23.44 +/- 14.73. Accumulative prevalence in children and young adult reached 74.7% of total patients. There was a trend that the subclinical infection rate raised gradually from 0-15 years group to over 45 years group. The percent of lymphocyte in 0-15 years group was significantly higher than other age groups, P = 0.039. The average time of virus shedding were 6.5 +/- 2.10 days (from 2 days to 12 days) , and there were no significant difference in diverse age groups, P = 0.272. 13 out of 95 (13.7%) patients presented complications related with A-H1N1 infection, and 4 of 6 patients complicated with pneumonia were in the 0-15 years group. CONCLUSION: The distribution of age in A-H1N1 infection is markedly different from seasonal influenza, with more cases in school children and young adults and fewer cases in older adults. Flu-like symptoms in children were apparent and pneumonia was the major complication in children.

Diagnostic value of glypican-3 in serum and liver for primary hepatocellular carcinoma.

AIM: To evaluate the diagnostic value of glypican-3 (GPC3) in serum and liver for primary hepatocellular carcinoma (HCC). METHODS: Serum levels of GPC3 and α-fetoprotein (AFP) were measured in 75 patients with primary HCC and 32 patients with liver cirrhosis. Expression of GPC3 and AFP in 58 HCC and 12 cirrhotic specimens was detected with immunohistochemical staining. RESULTS: When the cut-off value of serum GPC3 was set at 300 ng/L, its sensitivity and specificity for HCC were 47.0% and 93.5%, respectively. Among the 14 patients with HCC at stage according to the Barcelona Clinic Liver Cancer staging system, the serum GPC3 level was higher than 300 ng/L in 50% (7/14) patients, the serum AFP level was not ≥ 400 µg/L in any patient. Combined serum AFP and GPC3 significantly increased the sensitivity to the diagnosis of HCC. The GPC3 expression was detected in cytoplasm of HCC cells but not in hepatocytes and bile ducts of benign tumors. Among the 58 HCC patients, the GPC3 was expressed in 100% (28/28) patients with their serum AFP level ≥ 400 µg/L, and in 90% (27/30) patients with their AFP level < 400 µg/L, respectively. The GPC3 was weakly or negatively expressed in all paracarcinomatous and cirrhotic tissue samples. AFP positive HCC cells were only found in 1 out of the 58 HCC patients. CONCLUSION: GPC3 protein is a sensitive and specific serum marker for diagnosis of early HCC. Its expression in liver tissues can be used to discriminate tumor cells from benign hepatic cells.

[Detection of antigen-epitope-specific cytotoxic T lymphocytes in patients with hepatitis B virus infection by enzyme linked immunospot assay].

OBJECTIVE: To investigate the ability of secreting interferon-gamma (IFN-gamma) of the peripheral blood monocular cells (PBMC) stimulated by hepatitis B virus (HBV)-specific cytotoxic T lymphocyte (CTL) epitopes peptides and to analyze the difference of CTL immune response in patients with HBV infection. METHODS: Four HLA-A2-restricted HBV cytotoxic T lymphocyte epitopes [Tp: HBV polymerase 575-583 (FLLSLGIHL), Te1: envelope 28-39 (IPQSLDSWWTSL), Te2: envelope 183-191 (FLLTRILTI) and Tc: core 18-27 (FLPSDFFPSV)] were synthesized. Human leucocyte antigen (HLA)-A2 typing was detected by Flow cytometry. PBMCs which were isolated from patients with chronic hepatitis B(CHB), patients with chronic severe hepatitis B(CSH), subjects with past HBV infection(N1) and healthy blood donors (N2) were stimulated by the four HLA-A2-restricted HBV CTLs epitopes. Enzyme linked immunospot (ELISPOT) assay was used to detect the frequency of secreting IFN-gamma CTL in each group. RESULTS: (1) HLA-A2 typing: 20 of 44 patients with CHB (45.5%) were HLA-A2 positive, 10/18 (55.6%) in CSH and 6/10 (60%) in group N1 were HLA-A2 positive.10 healthy blood donors' HLA-typing was detected in the early study.(2) ELISPOT results: (1) The total responses to the four epitopes in CHB, CSH, N1 and N2 groups were 50% (10/20), 10% (1/10), 83.3% (5/6) and 10% (1/10), respectively. The response in N1 group was significantly higher than that in CSH group (chi(2) = 9.000, P = 0.008) and N2 group (chi(2) = 9.000, P = 0.008). (2) The CTL average magnitude response to Tp epitope, Te1 epitope, Te2 epitope and Tc epitope was also significantly higher in past HBV infection group (77 SFC/10(6) PBMC, 59 SFC/10(6) PBMC, 100 SFC/10(6) PBMC and 57 SFC/10(6) PBMC, respectively) than that of CSH group (10 SFC/10(6) PBMC, 0 SFC/10(6) PBMC, 0 SFC/10(6) PBMC and 20 SFC/10(6) PBMC respectively, all P < 0.01) and N2 group (15 SFC/10(6) PBMC, 0 SFC/10(6) PBMC, 22 SFC/10(6) PBMC and 30 SFC/10(6) PBMC respectively, all P < 0.01). CONCLUSION: This study indicates that the T cell immune response to HBV-specific epitopes might be detected either in patient with chronic HBV infection or with previous HBV infection. This response should be much higher in patients with past HBV infection, even the virus had been cleared for long time. These results demonstrate that HBV-specific CTL might play an important role in the clearance of the virus.

[The etiology and clinical manifestations of 70 patients with hand-foot-mouth disease].

OBJECTIVE: To analyze the etiology and clinical manifestations of 70 patients with hand-foot-mouth disease (HFMD). METHODS: The viral RNA from the pharynx swab samples were extracted and collected from 70 HFMD patients (of which, 60 cases were under 5 years old) to detect the 5'-UTR gene of EV, the Vp3-Vp1 genes of EV71 and Cox-A16 by utilizing the technique of RT-PCR. RESULTS: Of 70 HFMD patients, 30 cases (42.8%, 30/70) were positive in enterovirus nucleic acid, including EV71 and Cox-A16, pharynx swab sample testing. While, of the enteroviral pathogen RNA-positive 30 cases, EV71 accounted for 66.7% (20/30). The proportion of etiological positive of 39 samples collected within 4 days after onset was 66.7% (26/39), but the probability of 31 samples collected after more than 5 days after onset was 12.9% (4/31), the difference was significantly (chi(2) = 20.4, P < 0.01). CONCLUSION: HFMD patients were mainly infants and young children between 0-5 years old, but adults could also be attacked. The enterovirus detected was mainly EV71, and the pharynx swab samples should be collected within 4 days after onset to increase the related viral nucleic acid positive detection probability.

Upregulation of heme oxygenase-1 with hemin prevents D-galactosamine and lipopolysaccharide-induced acute hepatic injury in rats.

Heme oxygenase-1 (HO-1), the rate-limiting enzyme in heme catabolism, has been shown to be induced during oxidative injury, and its induction acts as an important cellular defense mechanism against such injuries. In this study, we examined the functional roles of HO-1 induction in a rat model of d-galactosamine (GalN) and lipopolysaccharide (LPS)-induced liver injury. We found that GalN/LPS treatment of rats produced severe hepatic injury, whereas upregulation of HO-1 by hemin pretreatment prevented rats from liver damage, as evidenced by decreased serum ALT, AST levels and ameliorated histological signs in the liver. Induction of HO-1 resulted in a significant decrease in hepatic malondialdehyde (MDA) contents, tumor necrosis factor-alpha (TNF-alpha) levels, iNOS/NO production, as well as the levels of caspase-3. In contrast, inhibition of HO activity by zinc protoporphyrin-9 (ZnPP, a specific inhibitor of HO) completely reversed HO-1-induced hepatoprotective effect. These data therefore suggested that HO-1 induction provided critical protection against GalN/LPS-induced liver injury, and the protection seemed to be mediated through the anti-oxidant, anti-inflammatory and anti-apoptotic functions.

[The significance of anti-soluble liver antigen/liver-pancreas in diagnosing and typing autoimmune hepatitis].

OBJECTIVE: To study the significance of anti-soluble liver antigen/liver-pancreas (anti-SLA/LP) in diagnosing and typing autoimmune hepatitis (AIH). METHODS: Six thousand patients with abnormal liver functions, who had their autoantibodies checked by immunofluorescent assay (IFA) and immune blotting assay, were reviewed retrospectively. RESULTS: Of these 6000 patients with liver diseases, 84 were diagnosed AIH. Eighteen patients, 0.3% of the 6000, patients with abnormal liver functions, were SLA/LP antibody positive, of which 17 were with AIH-III [2/17 with AIH /primary biliary cirrhosis (PBC) overlap syndrome], and 1 with chronic hepatitis B. Sensitivity and specificity of SLA/LP antibody in diagnosing AIH were 20.2% and 99.7% respectively, and the positive prediction value was 94.44%. The antinuclear antibody (ANA) titer in the AIH-III group was significantly lower than that of the AIH-I group (P < 0.05). The age of patients with anti-SLA/LP was higher (58.8% were over 50 years old) than those without this antibody (52.2% were 30 to 50 years old). There were no significant differences between the type III and type I AIH regarding gender, age, abnormal degree of liver function, PTA, IgG, liver cirrhosis rates and response to immunosuppressive therapy. CONCLUSION: Anti-SLA/LP is highly specific for diagnosing AIH. Comparing the clinical data of type III and type I AIH, we did not find significant differences between the two groups.

Effects of salvianolic acid a on oxidative stress and liver injury induced by carbon tetrachloride in rats.

In the present study, we investigated the hepatoprotective effects of salvianolic acid A, a novel antioxidant, against oxidative stress and acute liver injury induced by carbon tetrachloride (CCl(4)) in rats, and the mechanisms underlying its protective effects. Administration of CCl(4) to rats caused severe hepatic damage, as demonstrated by the significant increase in the levels of serum alanine aminotransferase, aspartate aminotransferase and classic histological changes including hepatocyte necrosis or apoptosis, haemorrhage, fatty degeneration, etc. Co-treatment with salvianolic acid A (20 mg/kg, intraperitoneally), a water-soluble extract from a Chinese traditional drug, Radix Salvia miltiorrhiza, significantly decreased CCl(4)-induced hepatotoxicity. Salvianolic acid A not only decreased serum alanine aminotransferase, aspartate aminotransferas levels and ameliorated histopathological manifestations in CCl(4)-treated rats, but also reduced oxidative stress, as evidenced by decreased reactive oxygen species production and malondialdehyde concentrations in the liver tissues, combined with elevated hepatic superoxide dismutase activity and gluthathione content. In addition, salvianolic acid A treatment remarkably reduced intrahepatic tumour necrosis factor-alpha concentrations and caspase-3 activities as compared with the CCl(4)-treated rats. The results suggested that treatment with salvianolic acid A provides a potent protective effect against acute hepatic damage caused by CCl(4) in rats, which may mainly be related to its antioxidative effect.

[A follow up study of total IgM, IgG, nucleoprotein and spike protein antibodies against severe acute respiratory syndrome (SARS) coronavirus in patients with SARS].

OBJECTIVE: To survey the dynamic changing and persistence of the special antibodies, including total IgM, IgG, nucleocapsid protein and spike protein antibodies, against severe acute respiratory syndrome coronavirus (SARS-CoV) in patients with SARS. METHODS: 146 cases, all clinically diagnosed as SARS with positive SARS-CoV IgG, were followed up. 362 serum samples were collected from the onset of the disease to 660 days afterward. Total IgM and IgG against SARS-CoV were tested with commercial ELISA kits. For recombinant nucleoprotein and spike protein, we developed an ELISA to test these two antibodies. RESULTS: Within 20 days of the onset, the positive rate of anti-SARS-CoV IgM was 46.5% (20/43); it reached a peak after 21 - 40 days (80.6%, 25/31). Then, the positive rate of IgM went down gradually to 8.2% (6/73) until 550 days after the onset. The patient's IgG positive rate was lower (34.9%, 15/43) than that of IgM within 20 days of the onset. Then it went up rapidly to 100%. It remained positive (98.6%, 70/71) until 600 - 660 days after the onset. When N-IgG and S-IgG were tested 40 days after the onset of the disease at three different times, the positive rate of N-IgG (92.5%, 37/40) was higher than that of S-IgG (67.5%, 27/40), but the two structure protein antibodies were always lower than the total IgG. CONCLUSIONS: In SARS patients with definite clinical and etiological diagnosis, the highest positive rate of the antibodies against SARS-CoV was found at 21 - 40 days after the onset. IgM disappeared almost 500 days (91.8%) after the onset. Total IgG positive rate could reach 100% and 98.6% and the positivity might persist nearly two years. So it is speculated that the total IgG antibody may be positive 3 to 5 years after infection, but it seems that N-IgG and S-IgG keep positive shorter in time than total IgG antibody.

[Clinical and laboratory characteristics of anti-soluble liver antigen/liver-pancreas (SLA/LP) autoantibody positive liver disease patients].

OBJECTIVE: To analyze the clinical and laboratory features of anti-soluble liver antigen/liver-pancreas (SLA/LP) autoantibody positive patients with abnormal liver functions. METHODS: From July 1999 to August 2004, 4928 serum samples from patients with abnormal liver functions (ALT >40 U/L) were collected. A series of autoantibody examinations were carried out. Clinical manifestations and laboratory findings of 8 patients with anti-SLA/LP autoantibody positive were reviewed. RESULTS: Among the 5500 serum samples, 8 cases (6 females and 2 males) with positive anti-SLA/LP autoantibodies were found with complete clinical information. The age of the patients was (27-76) years old. The case histories were from 2 years to 10 years. Of the 8 patients, 6 cases had liver cirrhosis and HBsAg-negative and anti-HCV-negative, active, 1 case had liver cirrhosis with HBsAg-positive, but HBVDNA negative; 1 case had liver cirrhosis and anti-HCV positive, but HCV RNA negative. The 8 cases were all ANA positive with titers of 31:320. Four cases were AMA positive and 2 among these 4 cases were M2 positive. The most frequent symptoms were fatigue, anorexia, nausea, jaundice, abdominal distention and edema of lower limbs. All patients had high hypergammaglobulinemia. CONCLUSION: Anti-SLA/LP autoantibody was at a low detection rate in the study with females in preponderance, Clinical and laboratory characteristics of the 8 cases were consistent with those of the autoimmune hepatitis (AIH). Testing for anti-SLA autoantibodies helps in the diagnosis of AIH in many patients who may otherwise be misdiagnosed.

Combined human growth hormone and lactulose for prevention and treatment of multiple organ dysfunction in patients with severe chronic hepatitis B.

AIM: To evaluate the efficiency and safety of combined recombinant human growth hormone (rhGH) and lactulose for treatment and/or prevention of multiple organ dysfunction in patients with chronic severe hepatitis B. METHODS: Forty-eight inpatients with chronic severe hepatitis B were randomly divided into rhGH group (n=28) and control group (n=20). In rhGH group, 4-4.5 IU of rhGH was injected intramuscularly once daily for 2-4 wk, and 100 mL of enema containing 30 mL of lactulose, 2 g of metronidazole and 0.9% saline was administered every 2 d for 2-4 wk. Their symptoms and complications were noted. Liver and kidney functions were analyzed by an Olympus analyzer. Serum GH, IGF-1, IGFBP1 and IGFBP3 were measured by ELISA. RESULTS: Clinical symptoms of 90% of these patients in rhGH group were obviously improved. The total effectiveness in rhGH group was better than that in control group (75% vs 40%, P<0.05). After 2- and 4-wk treatment of rhGH respectively, serum albumin (26.1+/-4.1 vs 30.2+/-5.3, 31.9+/-5.1 g/L), prealbumin (79.6+/-28.0 vs 106.6+/-54.4, 108.4+/-55.0 g/L), cholesterol (76.3+/-16.7 vs 85.6+/-32.3, 96.1+/-38.7 mg/dL), and IGFBP1 (56.8+/-47.2 vs 89.7+/-50.3 ng/mL after 2 wk) were significantly increased compared to control group (P<0.05). However, serum GH was decreased. The increase of serum IGF1 and IGFBP3 after rhGH treatment was also observed. CONCLUSION: rhGH in combination with lactulose may be beneficial to the prevention and treatment of multiple organ dysfunction in patients with chronic severe hepatitis.

[Immunological characteristics of primary biliary cirrhosis patients].

OBJECTIVE: To study the clinical significance of the immunological characteristics in patients with primary biliary cirrhosis (PBC). METHODS: 3000 patients with abnormal liver functions were examined for anti-nuclear antibodies (ANA), anti-mitochondrial antibodies (AMA), anti-smooth muscle antibodies (SMA) and anti-liver kidney microsomal antibody (LKM) using immunofluorescent assays (IFA). LKM-1, liver cytosolic-1 (LC-1), soluble liver antigen (SLA)/liver- pancreas antigen (LP) and subtype of AMA (M2, M4, M9) as well as ANA profile were detected by an immune blotting assay and an enzyme-linked immune absorbent assay (ELISA), respectively. Cytokines were tested by flow cytometry. RESULTS: Of the 3000 patients with liver diseases, 52 (1.7%) were diagnosed with PBC. All the PBC cases were positive for AMA and M2. 94% of them showed high titer of AMA (> or = 1:320), and in 79% of them M2 was >200 RU/L, and 78% of them were ANA positive. Three main fluorescent patterns of ANA seen were nuclear membrane, nuclear dots and centromere patterns. Sjogren's Syndrome A/B (SS-A/B), homogenous, nucleolar or nuclear granular patterns were seen in only a few patients. IgM, ALP and GGT in PBC patients were significantly higher than those in hepatitis B related liver cirrhosis patients. The levels of IL-6, IL-10, TNF-alpha and IFN-gamma in PBC patients were higher than in the normal controls. Among the 52 PBC patients, 5 had autoimmune liver disease overlap syndromes. Two of them were SLA/LP positive, indicated as AIH type III and PBC overlapping, and 1 was LKM-1 positive showing AIH type II overlapping PBC, and 2 had ANA positive and were identified as AIH and PBC by liver biopsy. CONCLUSION: The percentage of PBC in Chinese liver disease patients is about 1% to 2%. Most of the PBC patients have high levels of AMA and AMA-M2, IgM, ALP, GGT and several cytokines, indicating that abnormality of humeral and cellular immunity may be associated with the pathogenesis of PBC.