Objective To investigate the preventive and therapeutic effects of shuixianzi extracts on hyperlipidemia rats, and on fatty liver pathology. Methods Shuixianzi was homogenized and filtered. Then the filtrate was freeze-dried after centrifugation. The powder was just the extracts of shuixianzi. During the establishment of rat hyperlipidemic model, the extract was given at the same time. At the end of this experiment, the changes of blood lipid and liver pathology were observed. In therapeutic experiments, after the hyperlipidemia model was established, optimal dose of extract was given, then the changes of blood lipid and liver pathology were also observed and the levels of superoxide dismutase (SOD) and malondialdehyde (MIA) were tested. Results In preventive experiments, high dose of extracts of shuixianzi versus negative control could inhlbit both the increase of TC, TG, LDL-C and the drop of HDL-C. [TC: (3.23±0.01) vs. (6.56±0.01) mmol/L; TG:(2.33±0.01) vs. (4.12±0.02) mmol/L; LDL-C: (2.02±0.01) vs. (3.91±0.02) mmol/L; HDL-C: (0.98±0.01) vs. (0.76±0.01) mmol/L, all P＜0.01]. At the same time, the extracts could inhibit the pathological changes of fatty liver. In therapeutic experiments, extracts versus control could regulate the serum lipid levels [TC: ( 3.67 ± 0.31 ) vs. ( 6.33 ± 0.52 ) mmol/L; TG: ( 1.99 ±0.11) vs. (4.08±0.24) mmol/L; LDL-C: (1.57±0.12) vs. (3.78±0.14) mmol/L; HDL-C:(1.10±0.03) vs. (0.77±0.02) mmol/L, all P＜0.01] and could reverse fatty changes of liver in hyperlipidemic rats. At the same time the extracts versus control could also increase the activity of SOD [(276.3±26.8) vs. (165.4±16.7) U/mg, P＜0.01] and decrease the level of MDA [(3.67±1.23) vs. (7.45±2.33) nmol/mg, P＜0.01]. Conclusions The extracts of shuixianzi could prevent and treat the hyperlipidemia, inhibit the fatty pathological change of liver, and also have the antioxidant function.

Objective To evaluate the immunotoxicity effect of Liver targeting interferon (IFN -CSP) on mice.Methods Mice were randomly divided into five groups:low, middle and high dose of IFN-CSP, solvent control group(saline) and Positive control group (cyclophosphamide).They were injected subcutaneously for 2 weeks.Delayed hypersensitivity test was used to determine the cell immunefunction and plaque forming cell assay was used to determine the humoral immune function.Results There was no significant difference of the the index of immune organ and the ear swelling degree between IFN-CSP groups and control group.There was also no significant difference on hemolytic plaque test between them.Conclusion IFN-CSP has no significant effect on both cellular immunity function and humoral immune function of mice, this results will provides the basis for further safety evaluation.

Background and purpose:Checkpoint kinase 1 and 2 have been proposed to be potential therapeutic targets to sensitize cancers to radioor chemo-therapeutics. However, little is known about whether Chk1/2 is also a suitable target for sensitizing cancers to curcumin. In the present study, we investigated effects of Chk1/2 siRNA on curcumin-induced apopotosis in hepatoma cell line Huh7 and evaluated the effectiveness of Chk1/2 as a therapeutic target to potentiate human hepatoma to curcumin. Methods:Effect of curcumin on the cell cycle checkpoint-associated proteins was detected by Western blot. The knockdown efficacy of Chk1/2 siRNA was measured by RT-PCR and Western blot. Effect of Chk1/2 siRNA on curcumin-induced apoptosis in Huh7 cells was evaluated by DAPI staining. Effect of Chk1/2 siRNA on cell cycle distribution in curcumin-treated Huh7 cells was analyzed by flow cytometry. Results:Curcumin significantly inhibited phosphorylation of cell cycle checkpoint-associtaed proteins Chk1(S317), Cdc25C(S216) and Cdk1(Y15). Chk1 siRNA decreased Chk1 mRNA and protein by 95% and 92% and Chk2 siRNA decreased Chk2 mRNA and protein by 60% and 55% respectively as compared with negative control siRNA (P

Expression conditions of induction strategies for the cytoplasmic inclusion bodies (IBs) production of liver targeted interferon IFN-CSP by recombinant Escherichia coli (E. coli) BL21 (DE3) were optimized in shake-flask cultures in this study. The factors of the optimized protocol included in the present study were pH, inducer IPTG (isopropyl beta-D-thiogalactoside) concentration, culture growth temperature, incubation time and induction point. The effects of those factors were investigated by 'single variable at a time' method, aimed to analyze characterization of the recombinant strain. Orthogonal experimental design was further used to optimize the above critical factors for IFN-CSP production. According to the expression optimization result, it was confirmed that the main influence factors were cell density and induction temperature. The IFN-CSP gene expression optimized conditions were: pH value of the culture medium was 6.0, culture temperature 37 degrees C, adding IPTG to final concentration 0.4 mmol/L when the recombinant strain growth density OD600 achieved 0.8 and induction time 4 h. At this point, the IBs represented 74.3% of the total cellular protein. Compared with the non-optimized condition, IFN-CSP production obtained in optimized induction strategies were increased by approx. 1.2-fold. The optimized induction strategy yielded 688.8 mg/L of IFN-CSP, providing experimental data to study the biology activity and productive technology of IFN-CSP.
Biotechnology ; methods ; Cell Culture Techniques ; methods ; Culture Media ; chemistry ; Escherichia coli ; metabolism ; Gene Expression ; Interferons ; biosynthesis ; Liver ; Temperature

Background and purpose: Checkpoint kinase 1 and 2 have been proposed to be potential therapeutic targets to sensitize cancers to radio- or chemo-therapeutics. However, little is known about whether Chk1/2 is also a suitable target for sensitizing cancers to curcumin. In the present study, we investigated effects of Chk1/2 siRNA on curcumin-induced apopotosis in hepatoma cell line Huh7 and evaluated the effectiveness of Chkl/2as a therapeutic target to potentiate human hepatoma to curcumin. Methods: Effect of curcumin on the cell cycle checkpoint-associated proteins was detected by Westem blot. The knockdown efficacy of Chk1/2 siRNA was measured by RT-PCR and Westem blot. Effect of Chk1/2 siRNA on curcumin-induced apoptosis in Huh7 cells was evaluated by DAPI staining. Effect of Chk1/2 siRNA on cell cycle distribution in curcumin-treated Huh7 cells was analyzed by flow cytometry. Results: Curcumin significantly inhibited phosphorylation of cell cycle checkpoint-associtaed proteins Chk1(S317), Cdc25C(S216) and Cdk1(Y15). Chk1 siRNA decreased Chk1 mRNA and protein by 95% and 92% and Chk2 siRNA decreased Chk2 mRNA and protein by 60% and 55% respectively as compared with negative control siRNA (P<0.01). Inhibition of Chk1, but not Chk2, increased apoptotic rate from (21.3±1.8)% to (29.5±2.6)% (P<0.05). Neither Chk1 nor Cbk2 siRNA had any impact on cell cycle distribution in Huh7 cells induced by curcumin. Conclusion: Chk1 siRNA sensitized Huh7 cells to curcumin-induced apoptosis, suggesting that Chk1 is a potential therapeutic target to sensitize human hepatoma to curcumin.

The present study is to elucidate the mechanisms underlying Gleevec-induced apoptosis of chronic myeloid leukemia (CML) K562 cells in vitro. The apoptotic cell death and cell cycle distribution after Gleevec treatment and the effect of PDCD4 siRNA on Gleevec-induced apoptosis of K562 cells were analyzed by flow cytometry. The effect of Gleevec on p-Crkl, caspase-3, PARP and PDCD4 protein levels, and the knockdown efficacy of PDCD4 siRNA were detected by Western blotting. The results showed that Gleevec dramatically suppressed the phosphorylation level of Crkl in a dose-dependent manner and induced significant apoptosis and G0/G1 cell cycle arrest of K562 cells in time- and dose-dependent manners. In addition, Gleevec activated caspase-3 and its downstream substrates PARP, and the caspase pan inhibitor Z-VAD-FMK (50 micromol x L(-1)) markedly reduced Gleevec-induced apoptosis from 47.97% +/- 10.56% to 31.05% +/- 9.206% (P < 0.05). Moreover, Gleevec significantly increased the protein expression of programmed cell death 4 (PDCD4). PDCD4 knockdown by siRNA reduced Gleevec-induced apoptosis from 46.97% +/- 14.32% to 42.8% +/- 11.43%. In summary, Gleevec induced apoptosis in K562 cells via caspase-3 activation.

Hepatitis c virus (HCV) infection has become one of the global public health problem,while there is no vaccine to prevent HCV infection, the so-called "cocktail" therapy that use a combination of drugs targeting multiple steps in the HCV infection cycle could achieve better curative effect. the process of HCV entering into host cell is the important step of drug intervention, in which HCV envelope protein El and E2, Host cell factors including Heparan sulfate(HS), CD81, scavenger receptor class B type I (SR-BI), Occludin (OCLD), Claudin (CLDN), low densitity lipoprotein receptor (LDLR), dendritic cell-specific ICAM-3-grabbing nonintegrin (DC-SIGN), Liver/lymph node specific ICAM-3-grabbing integrin(L-SIGN), trans- ferrin receptor 1 (TfR1) and so on play a important role. The virus and the host factors can be used as targets of hcv entry inhibitors many studies have shown that as novel and promising compounds, HCV entry inhibitors combinating with other drugs can be more effective in the treatment of HCV, this paper have re- viewed targets and inhibitors of HCV enterring into host cell since 1990s.
Animals ; Antiviral Agents ; pharmacology ; Hepacivirus ; drug effects ; physiology ; Hepatitis C ; genetics ; metabolism ; virology ; Humans ; Receptors, Virus ; genetics ; metabolism ; Viral Envelope Proteins ; genetics ; metabolism ; Virus Internalization ; drug effects

Objective To investigate the anti-HBV molecular mechanisms of liver targeting interferon ( IFN-CSP ) in Balb/c-HBV transgenic mice.Methods Balb/c-HBV transgenic mice were randomly divided into 3 groups.Control group (treated with physiological saline), IFN α2b group (treated with 103 U/g IFN α2b), IFN-CSP group (treated with 102 U/g IFN-CSP).Another group of the non-transgenic mice were used as the Normal group.Each mouse was intramuscular injected with 50 μL dose once a day for 4 weeks.Total RNA of mice liver were extracted, and STAT1, STAT2, IRF-9, OAS1 gene expression of JAK-STAT signaling pathway were analyzed by real-time PCR.Results IFN α2b and IFN-CSP can significantly up regulate the expression of STAT1, STAT2, IRF-9, OAS1 gene of JAK-STAT signaling pathway (P<0.01).The induce effects of IFN-CSP on STAT1, STAT2, IRF-9, OAS1 were significantly better than that of IFN α2b (P<0.05).Conclusion The anti-HBV molecular mechanisms of liver targeting interferon (IFN-CSP) in Balb/c-HBV transgenic mice maybe related to regulate the expression of STAT1, STAT2, IRF-9, OAS1 gene of JAK-STAT signaling pathway.These results will lay a basis for the application of recombinant liver-targeting interferon.

BACKGROUND:The emergence of a large number of hidden blood loss during perioperative period of intertrochanteric fracture in the elderly not only increases the risk of perioperative period and complications, but also affects the postoperative recovery of joint function. At present, there is no relevant report about nutritional status and the hidden blood loss before surgery in and outside China.
OBJECTIVE:To identify the effect of nutritional status on preoperative hidden blood loss in elderly patients with intertrochanteric fracture.
METHODS:183 elderly patients with fresh and initial femoral intertrochanteric fracture were included. Laboratory serological examinations on admission and preoperation were completed. By mini nutritional assessment, patients were randomly divided into normal-nourishment group, malnourishment at risk group, and malnourishment group. The original blood volume and preoperative hidden blood loss were calculated depending on height, weight, hematocrit on admission and preoperation. According to the proportion of mean preoperative hidden blood loss on the original blood volume, patients were divided into low and high hidden blood loss groups. We compared preoperative hidden blood loss, and their proportion on the original blood volume and the preoperative incidence of high hidden blood loss, and analyzed the correlations between preoperative high hidden blood loss and preoperative nutritional status.
RESULTS AND CONCLUSION:(1) The nutritional status of elderly intertrochanteric fracture patients measured by mini nutritional assessment score was that the number of patients was 48 cases (26%) in normal-nourishment group, 64 cases (35%) in the malnourishment at risk group, and 71 cases (39%) in the malnourishment group. There were no obvious differences in the preoperative complications between any two groups (P>0.05). (2) Thirty-eight cases affected high hidden blood loss. The mean preoperative hidden blood loss was 260.43 mL. The proportion of preoperative hidden blood loss to the original blood volume was 6%. (3) The preoperative hidden blood loss, their proportion on the original blood volume and the incidence of high hidden blood loss were significantly higher in the malnourishment at risk group and malnourishment group than in the normal-nourishment group. Paired comparison showed significant differences (P<0.05). (4) Results confirmed that preoperative hidden blood loss, their proportion on the original blood volume and the incidence of high hidden blood loss gradual y increased with deterioration of nutritional status. The nutritional status is an important factor influencing the occurrence of preoperative hidden blood loss, and can be used as an important index for judging the high hidden blood loss and prognosis in elderly patients with intertrochanteric fracture.

In the present study, the total RNA was extracted from three instar larvae of Musca domestica, the cDNA sequence encoding the ORF of cecropins was amplified by RT-PCR, and the target fragment was further sequenced after being cloned into T vector pUCm-T. Then, the cDNA sequence of the mature cecropins was amplified by PCR with recombinant plasmid pUCm-T/cecropin as template, the N-terminal rare codon GGA of E. coli was changed to the favorable codon GGC,and a Asn codon AAC was added in front of the stop coden TAA in the C- terminus. This mutant gene designated as mCecropin was then ligated with the fusion expression vector pGEX-4T-1. After restriction analysis and DNA sequencing, the positive recombinant plasmid pGEX-4T-1/mCecropin was transformed to different strains of E. coli cells and the fusion protein was expressed after IPTG induction. The fusion protein was assayed by SDS-PAGE and the E. coli BL21(DE3) cell was chosen as the host cell for the expression of the fusion protein. The expressed fusion protein GST-mCecropin was purified by GSTrap affinity coloum and the GST marker was then cleaved by thrombin. In this way, the fusion protein mCecropin with antibacterial activity was obtained after purification with HiTrap benzamidine column.