ABSTRACT:In this study ,we elucidated the predictors of progression to liver failure during severe acute exacerbation .We analyzed 69 consecutive patients with severe acute exacerbation of chronic hepatitis B for clinical outcome and factors that influ‐enced the development of liver failure ,including viral genotype ,PC (G1896A) and BCP (A1762T/G1764A) mutants .Thirty‐three (47 .8% ) severe acute exacerbation patients progressed to liver failure .Multivariate analysis identified serum bilirubin (TB>256 μmol/dL ,P=0 .008) and prothrombin activity (PTA<40% ,P<0 .001) as significant determinants of progression to liver failure .HBeAg negativity (P=0 .065) and PC mutant (P=0 .090) were associated with the progression to hepatic de‐compensation .Serum total bilirubin ,prothrombin activities ,HBeAg status and PC mutant were predictors of clinical outcome in patients with severe acute exacerbation of chronic hepatitis B .

To study the chemical constituents of Nervilia fordii (Hance) Schltr., various chromatographic methods were used, including D101 macroporous resin, silica gel, ODS and preparative HPLC chromatographic techniques. A new labdane diterpenoid glycoside named as nervilifordoside A was isolated from 60% EtOH extract of Nerviliafordii. The structure of compound 1 was elucidated as 12, 17-epoxy-3-hydroxy-17-methoxy-labdan-13-en-16, 15-olide 3-O-alpha-rhamnopyranosyl-(1 --> 2)-O-beta-glucopyranoside on the basis of HR-MS, 1D and 2D NMR spectroscopic data as well as chemical methods.

Objective To investigate the protective effects and underlying molecular mechanisms of hydrogen (H2) on high glucose-induced poly (ADP-ribose) polymerase-1 (PARP-1) dependent cell death (PARthanatos) in primary rat Schwann cells. Methods Cultured primary rat Schwann cells were randomly divided into five groups: blank control group (C group), H2 control group (H2 group), high osmotic control group (M group), high glucose treatment group (HG group), and H2 treatment group (HG+H2 group). The cells in H2 group and HG+H2 group were cultured with saturated hydrogen-rich medium containing 0.6 mmol/L of H2, and those in three control groups were cultured with low sugar DMEM medium containing 5.6 mmol/L of sugar, and the cells in HG and HG+H2 groups were given 44.4 mmol/L of glucose in addition (the medium containing 50 mmol/L of glucose), the cells in C group and H2 group were given the same volume of normal saline, and the cells in M group were given the same volume of mannitol. Cytotoxicity was evaluated using lactate dehydrogenase (LDH) release rate assays after treatment for 48 hours in each group. The contents of peroxynitrite (ONOO-) and 8-hydroxy-2-deoxyguanosine (8-OHdG) reflecting oxidative stress injury and DNA damage were detected by enzyme linked immunosorbent assay (ELISA). Poly (ADP-ribose) (PAR) protein expression was analyzed by Western Blot, and immunofluorescence staining was used to determine the nuclear translocation of the apoptosis-inducing factor (AIF). Results The cytotoxicity in HG and HG+H2 groups was significantly increased as compared with that of C group [LDH release rate: (61.40±2.89)%, (42.80±2.32)% vs. (9.92±0.38)%, both P < 0.01], the levels of ONOO- and 8-OHdG were markedly elevated [ONOO- (ng/L): 853.58±51.00, 553.11±38.66 vs. 113.56±14.22; 8-OHdG (ng/L): 1 177.37±60.97, 732.06±54.29 vs. 419.67±28.77, all P < 0.01], and the PAR protein expression was up-regulated (A value: 0.603±0.028, 0.441±0.010 vs. 0.324±0.021, both P < 0.01). The cytotoxicity, the levels of ONOO- and 8-OHdG, and PAR expression in HG+H2 group were significantly lower than those of the HG group (all P < 0.01). There were no significant differences in above parameters between H2 group as well as M group and C group. It was shown by immunofluorescence that AIF was expressed in the cytoplasm in C group, H2 group and M group, AIF was expressed in the whole cell in HG group, and the expression in the nucleus was particularly increased. A small amount of AIF expression was found in the nucleus of HG+H2 group, which indicated that high glucose could promote the AIF nuclear translocation, and that hydrogen-rich medium could prevent the process of translocation. Conclusions High glucose levels could enhance DNA damage that enhance PARthanatos in primary rat Schwann cells. However, H2 can not only reduce DNA damage of injured cells, but also inhibit the special death process, reduce the cell toxicity, all of which have protective effects.

ObjectiveTo establish separate and appraisal methods of murine bone marrow mesenchymal stem cells (MSCs) and optimize the suitable conditions inducting MSCs directional differentiating into adipocytes in vitro.MethodsThe differential adherence to plastic was employed to separate MSCs. CFU-f and successive CFU-f cultures were employed to characterize the potent of proliferation and self-renewal of MSCs. The different adipogenic medium was used as induction for the differentiation of MSCs into adipocytes. The differentiated cells were identified by oil red O immunohistochemistry stain.ResultsThe purified MSCs showed the morphology of fibroblasts. It was found that the number of CFU-f formation depended on the planted number of MSCs. It showed a good relationship. Small type colony of CFU-f had little potent to re-clone, but almost 90% big type colony of CFU-f had the potent to regenerate CFU-f. The MSCs could directionally differentiated into adipocytes induced by different adipogenic medium. But more than 96% MSCs differentiated into mature adipocytes when induced by combined with dexamethasone (DM), 1-methy-3-isobutylxanthine (IBMX), insulin (IS) and indomethacin (ID).ConclusionThe purified MSCs can be harvested by method of differential adherence to plastic, and these MSCs have the potent of proliferation and self-renewal. Moreover, more than 96% MSCs can differentiate into mature adipocytes when induced by combined with DM, IBMX, IS and ID.

To study the chemical constituents of Nervilia fordii (Hance) Schltr., various chromatographic methods were used, including D101 macroporous resin, silica gel, ODS and preparative HPLC chromatographic techniques. A new labdane diterpenoid glycoside named as nervilifordoside A was isolated from 60% EtOH extract of Nerviliafordii. The structure of compound 1 was elucidated as 12, 17-epoxy-3-hydroxy-17-methoxy-labdan-13-en-16, 15-olide 3-O-alpha-rhamnopyranosyl-(1 --> 2)-O-beta-glucopyranoside on the basis of HR-MS, 1D and 2D NMR spectroscopic data as well as chemical methods.
Chromatography, High Pressure Liquid ; Molecular Structure ; Orchidaceae ; chemistry ; Plants, Medicinal ; chemistry

OBJECTIVETo investigate the effect of oligochitosan in promoting intestinal absorption of protoberberine alkaloids in extracts from Corydalis saxicola total alkaloids.

METHODThe in vitro single-pass intestinal perfusion model in rats was established to study the changes in absorption kinetic parameters of dehydrocavidine, berberine hydrochloride and palmatine chloride in C. saxicola total alkaloids after the addition of different concentrations oligochitosan and evaluate the effect of oligochitosan in promoting intestinal absorption of the drugs.

RESULTThe concentration of oligochitosan had different effects on the absorption rate constant (Ka) and apparent permeability coefficient (Peff) of the three active component in rat intestines. Ka and Peff in 0.5% oligochitosan group significantly increased, indicating a stronger effect in promoting the absorption.

CONCLUSIONOligochitosan has a certain effect in promoting the intestinal absorptions of protoberberine alkaloids in C. saxicola total alkaloids.

Animals ; Berberine Alkaloids ; administration & dosage ; pharmacokinetics ; Chitin ; administration & dosage ; analogs & derivatives ; Corydalis ; chemistry ; Drugs, Chinese Herbal ; administration & dosage ; pharmacokinetics ; Intestinal Absorption ; drug effects ; Intestines ; drug effects ; metabolism ; Male ; Rats ; Rats, Sprague-Dawley

As an important part of traditional Chinese medicine (TCM), mineral medicine plays an irreplaceable role. However, little has been reported on its species and valence state of heavy metals and deleterious elements, and also the relevance to pharmacological effect and toxicology. The present paper, in a new perspective, summarized the determination of the species and valence state of heavy metals and deleterious elements in recent years, discussed the progress of the pharmacological effect and toxicology, and prospected for future study which might provide reference for mineral medicine.
Animals ; Drug Contamination ; statistics & numerical data ; Drug Therapy ; Humans ; Medicine, Chinese Traditional ; Metals, Heavy ; analysis ; toxicity ; Minerals ; analysis ; pharmacology

Objective To investigate the correlation between serum level of visfatin and type 2 diabetes mellitus (T2DM). Methods Impaired glucose regulation (IGR) group consisted of 40 patients,T2DM group consisted of 106 patients, while control group consisted of 86 subjects. The serum visfatin levels of all groups were detected by enzyme linked immunosorbent assay (ELISA). Results Fasting serum visfatin levels in IGR group and T2DM group were significantly higher than those in control group [(19.93 ±6.89) μg/L and (29.53 ± 11.33) μg/L vs. (16.12 ± 5.24) μ g/L, P ＜ 0.01]. Fasting serum visfatin levels positively correlated with waist-to-hipratio (WHR) (r = 0.161, P ＜ 0.05); significantly positively correlated with triacylglycerol (TG), low density lipoprotein cholesterol (LDL-C), glycosylated hemoglobin (HbA1c),fasting plasma glucose (FPG), fasting insulin (FINS), lg Homeostasis model assessment-insulin resistance index (HOMA-IR) (r = 0.189,0.266,0.643,0.574,0.285,0.526,P ＜ 0.01), and significantly negatively correlated with high density lipoprotein cholesterol (HDL-C)(r =-0.377,P ＜0.01). When visfatin was analyzed as a dependent variable in multiple linear regression, HbA1c (β = 0.512, P = 0.000), lg HOMA-IR (β= 0.172, P = 0.026), WHR (β = 0.119, P = 0.036) were into the equation. Conclusion Visfatin may be involved in glucose and lipid metabolism regulation, and closely related with insulin resistance; visfatin may be a risk factor for T2DM.

The inhibition effects of the extracts of five Verbenaceae herbs, Clerodendrum fortunatum L. , Clerodendrum japonicum(Thunb. )Sweet, Clerodendrum philippinum Schauer var. simplex Moldenke, Callicarpa longissima(Hemsl. )Merr. and Pygmaeopremna herbacea, were investigated by cell hemolysis model in vitro on classical and alternative complement activation pathways. The water extracts of Pygmaeopremna herbacea, the water extract and the ethanol extract of Clerodendrum fortunatum L. , the water extract and the ethanol extract of Callicarpa longissima(Hemsl. )Merr. , the ethanol extracts of Clerodendrum japonicum(Thunb. )Sweet and Clerodendrum philippinum Schauer var. simplex Moldenke showed inhibition cell hemolysis effects on the classical pathway. Their CH50 values were 0. 092±0. 008, 0. 074±0. 008, 0. 088±0. 006, 0. 134±0. 017, 0. 123±0. 010, 0. 380±0. 080, and 0. 200±0. 015 g/L, respectively. The water extracts of Pygmaeopremna herbacea and Callicarpa longissima(Hemsl. )Merr. and the ethanol extract of Clerodendrum fortunatum L. showed inhibition cell hemolysis effects on alternative pathway. Their AP50 values were 0. 533±0. 033, 0. 758±0. 031, and 0. 362±0. 029 g/L, respectively. Five Verbenaceae herbs appear good anticomplementary effects in vitro. The ethanol extract of Clerodendrum fortunatum L. showed the best inhibitory activity.

OBJECTIVETo characterize genotypic resistance within HBV RT region in chronic hepatitis B (CHB) patients with nucleos(t)ide analogue (NA) treatment.

METHODSSerum samples of 229 CHB patients with NA treatment were obtained. Full-length HBV RT sequences were amplified, sequenced and analyzed, on the following NA resistant (NAr) mutations belonging to different NAr pathways.

RESULTSAmong 229 HBV isolates, 14.41% (33/229) and 85.59% (196/229) were genotype B and C, respectively; and the patients with HBV genotype C may be more susceptible to develope resistant mutations than patients with HBV genotype B(chi2 = 2.95, P < 0.05). NAr mutations were detected in 63 CHB patients. Mutations were not found at rtI169, rtT184, rtA194 or rtS202. RtM204 mutations were detected at the highest frequency among 63 mutants (40/63, 63.49%) and found to display 11 combination mutation patterns, in which rtM204I were associated with rtL80I/V and rtL180M, and rtM204V were associated with rtL1l80M, respectively. Conclusions There are complicated mutation patterns in the HBV RT region for chronic hepatitis B (CHB) patients with nucleos(t)ide analogue (NA) treatment. RtM204V/I mutation was the highest.

Adolescent ; Adult ; Aged ; Antiviral Agents ; therapeutic use ; Female ; Hepatitis B virus ; drug effects ; enzymology ; genetics ; Hepatitis B, Chronic ; drug therapy ; virology ; Humans ; Male ; Middle Aged ; Mutation ; drug effects ; Nucleosides ; therapeutic use ; Nucleotides ; therapeutic use ; RNA-Directed DNA Polymerase ; genetics ; metabolism ; Viral Proteins ; genetics ; metabolism ; Young Adult