ObjectiveTo discuss the relationship between the cross-reactivity of antibody against the hypervariable region 1 ( HVR1 ) of hepatitis C virus and early viral response ( EVR ) in patients undergoing antiviral therapy.MethodsBy ELISA and HCV HVR1 antibody cross-reactivity matrix reagent,the differences of anti-HCV hypervariable region antibody were tested in baseline serum from 46 patients with chronic hepatitis C before antiviral therapy.HCV genotyping and HCV RNA were analyzed by RT-PCR method.The HCV RNA assay was done at three time points:before treatment,pegylated interferon in combination with ribavirin therapy for 12 and 48 weeks.ResultsIn 46 cases of chronic hepatitis C patients,there were 16 cases with HCV 2a type,30 cases with l b and 33 patients obtained EVR.The EVR incidence of type 2a[ 93.8％ ( 15/16 ) ] was higher than that of type 1 b[ 60.0％ (18/30),x2 =4.316,P ＜ 0.05 ].In the EVR group of type 1b chronic hepatitis C patients,the positive number of average multi-target HVR1 antigen was ( 12 ± 4),which was significantly higher than that in the Non-EVR patients [ (7 ± 5 ),t =2.797,P ＜0.01 ].Bv the Fisher exact test,it was showed that patients'serum response to HVR1 antigens numbered 001,003,009,013,016 were higher in EVR group than those in non-EVR group,with statistically significant (P ＜ 0.05 ).ConclusionThe cross-reactivity of HVR1 antibody may play an important role in predicting the effectiveness of antiviral therapy.

OBJECTIVETo investigate the effects of shikonin on the proliferation, expression of CXCR4 and the migratory responses to CXCL12 in colorectal carcinoma cell line SW480.

METHODSThe proliferation of SW480 cells was assessed by MTT assay. Cell surface expression of CXCR4 was determined by flow cytometry. The migratory ability was determined by Transwell.

RESULTSShikonin inhibited the proliferation of SW480 cells in time- and concentration-dependent manner. The expression rate of CXCR4 in SW480 cells was 99.1%. After application of shikonin 0.01 micromol/L, 0.1 micromol/L and 1.0 micromol/L for 24 h, the expression rate of CXCR4 decreased to 76.0%, 59.1% and 35.5% respectively (F=1098.041, P <0.001), and the CXCL12-induced SW480 cell migratory inhibition rate was 25.2%, 38.5% and 55.7% respectively (F=48.970, P <0.001).

CONCLUSIONBesides having inhibiting tumor cell proliferation effect, Shikonin may also play a role in anti-metastasis via down-regulating the expression of CXCR4 and reducing the CXCL12-induced migratory response in colorectal carcinoma cell.

Cell Line, Tumor ; Cell Proliferation ; drug effects ; Chemokine CXCL12 ; metabolism ; Down-Regulation ; Humans ; Naphthoquinones ; pharmacology ; Receptors, CXCR4 ; metabolism

OBJECTIVETo study the changes of TLR2 and TLR4 on peripheral blood mononuclear cells (PBMCs) and their role in the pathogenesis of chronic hepatitis B and chronic severe hepatitis B.

METHODSThe expressions of TLR2 and TLR4 on 10000 CD14+ PBMCs were determined by flow cytometry in 30 healthy controls, in 31 patients with chronic hepatitis B and in 30 patients with chronic severe hepatitis B. The level of serum tumor necrosis factor alpha (TNF alpha) was determined by ELISA. The differences of expression of TLR2 and TLR4 on PBMCs and serum TNFalpha among the three groups of study subjects were determined by Student-t test. The correlations between TLR2, TLR4 and TNF alpha were determined by linear correlation test.

RESULTSThe values of mean fluorescence intensity (MFI) of TLR2 on PBMCs of the healthy controls, patients with chronic hepatitis B and patients with chronic severe hepatitis B groups were 21.5+/-2.7, 39.0+/-4.1, and 47.7+/-21.4; TLR4 of those groups was 2.3+/-1.1, 3.7+/-2.3, and 6.9+/-4.1. The serum TNF alpha(ng/L) of the respective groups was 53.8+/-38.1, 164.3+/-89.9, and 359.8+/-140.0. There was a gradual increase of these values from the group of healthy controls to the group of patients with chronic hepatitis B and patients with chronic severe hepatitis B. No significant positive correlations between TLR2, TLR4 and serum TNFalpha were found.

CONCLUSIONTLR2 and TLR4 may have a role in the pathogenesis of chronic hepatitis B and chronic severe hepatitis B.

Adult ; Aged ; Case-Control Studies ; Female ; Hepatitis B, Chronic ; blood ; Humans ; Male ; Middle Aged ; Monocytes ; metabolism ; Toll-Like Receptor 2 ; metabolism ; Toll-Like Receptor 4 ; metabolism ; Tumor Necrosis Factor-alpha ; blood

OBJECTIVESTo study the relationship between intra-hepatic levels of regulated on activation, normal T-cell expressed and secreted (RANTES) and the disease severity and liver inflammatory degrees in patients with chronic hepatitis B and the possible mechanism of the changes of intra-hepatic levels of RANTES.

METHODSThe expression of RANTES of the livers was studied using immunohistochemical stainings and morphometric quantitative measurements in liver specimens from 10 normal subjects and 64 patients with chronic hepatitis B with different degrees of liver inflammation and different clinical severity. The expressions of RANTES protein and mRNA in cell line HepG2, HepG2.2.15 and HepG2 treated with 10 ng/ml TNFa at different times were quantified by ELISA and one-step RT-PCR.

RESULTSThe expression of RANTES of the livers in patients was significantly higher than that in the normal controls. Hepatic RANTES levels increased significantly and the increases were parallel to the increases of the severity of the hepatitis, from mild, moderate to severe hepatitis (the positive units were 3.7+/-1.5, 15.6+/-6.9, 24.0+/-4.0, 37.9+/-11.1, respectively) and from G0 degree to G4 degrees of liver inflammation (the positive units were 3.7+/-1.5, 15.0+/-5.7, 21.6+/-5.9, 30.3+/-8.2, 40.9+/-12.3, respectively). The expressions of RANTES protein and mRNA of HepG2.2.15 were higher than that of HepG2. RANTES protein and mRNA were induced in HepG2 by TNFa.

CONCLUSIONRANTES may have an important role in the pathogenesis of chronic hepatitis B. The elevation of hepatic RANTES may be caused by hepatitis B virus and TNFa.

Adolescent ; Adult ; Case-Control Studies ; Chemokine CCL5 ; metabolism ; Female ; Hep G2 Cells ; Hepatitis B virus ; Hepatitis B, Chronic ; metabolism ; Humans ; Liver ; metabolism ; Male ; Middle Aged ; Tumor Necrosis Factor-alpha ; Young Adult

The effects of pretreatment of supercritical carbon dioxide (SC-CO2) on the supramolecular structure of cellulose and the cellulase catalyzed reaction were investigated. The cellulase activity was not affected when it was treated with SC-CO2 at 10MPa and at 50 degrees C for 30 min. But when the cellulase was treated by SC-CO2 in the presence of cellulose, the catalytic activity of the cellulase was lost. The cellulose pretreated with or without cellulase under the same SC-CO2 condition was then hydrolyzed with tresh crude cellulase. The final reducing sugar yield from the hydrolysis of the cellulose pretreated with cellulase was higher than that of the cellulose pretreated without cellulase. It was also found that the improvement of the enzymolysis had a direct relevance with the amount of cellulase used during the SC-CO2 pretreatment. The moisture content of cellulose before SC-CO2 pretreatment had an obvious influence on the subsequent enzymolysis. When the moisture content of cellulose was 60% (W/W), the reducing sugar yield was higher than when the moisture content was over 100% (W/W). The FT-IR spectra showed that the structure of the cellulose pretreated with cellulase under the SC-CO2 condition was different from that of the cellulose pretreated without cellulase. In the presence of the enzyme, the strength of the hydrogen bonds and the I beta phase at 710cm(-1) in the crystalline cellulose was weakened. These results suggest that the change in the cellulose structure induced by the SC-CO2 treatment favous the subsequent enzymolysis.
Carbon Dioxide ; chemistry ; Cellulase ; chemistry ; metabolism ; Chromatography, Supercritical Fluid ; methods ; Spectroscopy, Fourier Transform Infrared

OBJECTIVETo study the clinical characteristics of ceftriaxone-associated biliary pseudolithiasis in children with renal diseases.

METHODThree children with renal diseases developed biliary pseudolithiasis when they were treated with ceftriaxone. Their clinical and laboratory data were retrospectively analyzed.

RESULTSCase one was an 11-year-old boy. The initial diagnosis was primary nephrotic syndrome. Ceftriaxone was administered intravenously at a dose of 2 g/d [50 mg/(kg * d)] for gastroenteritis. After that the boy complained of nausea and loss of appetite. Abdominal sonogram obtained on day 3 of ceftriaxone therapy revealed gallbladder sludge. After cessation of ceftriaxone treatment, symptoms and ultrasound abnormalities gradually disappeared, with complete sonographic resolution after 16 days. Case two was a 10-year-old boy. The primary diagnosis was post-streptococcal glomerulonephritis with acute renal failure. The child was treated with 1.5 g/d [30 mg/(kg * d)] intravenous ceftriaxone for gastroenteritis. After that, the boy complained of nausea and abdominal pain with positive Murphy's sign. Gallstone was detected by ultrasonographic examination on day 6 of ceftriaxone therapy. After cessation of ceftriaxone treatment, symptoms and sonographic abnormalities gradually disappeared, with complete sonographic resolution after 18 days. Case three was a 12-year-old boy. The primary diagnosis was nephrotic syndrome. He was treated with 2 g/d [40 mg/(kg.d)] ceftriaxone for gastroenteritis. Gallbladder lithiasis was detected 17 days after the initiation of ceftriaxone therapy (3 days after cessation of ceftriaxone treatment). Gallbladder sonogram was found to be normal two months after the discontinuation of the therapy.

CONCLUSIONSBiliary pseudolithiasis occurred in 3 cases with renal diseases receiving low doses of ceftriaxone. The risk of developing ceftriaxone-associated biliary pseudolithiasis might increase in patients with renal diseases who are treated with ceftriaxone.

Anti-Bacterial Agents ; adverse effects ; therapeutic use ; Ceftriaxone ; adverse effects ; therapeutic use ; Child ; Cholecystolithiasis ; chemically induced ; Humans ; Kidney Diseases ; complications ; drug therapy ; Male ; Retrospective Studies

In order to study the distribution of virulence genes of Listeria monocytogenes (Lm) in Hebei Province,29 virulence genes of Lm,including Listeria monocytogenes pathogenicity islands Ⅰ (LIPI-Ⅰ:prfA,plcA,plcB,hlyA,mpl and actA),10 internalins genes (inlA,inlB,inlC,inlD,inlE,inlF,inlG,inlH /C2,inlI and inlJ) and the other 13 virulence-associated genes (bsh,srtA,iap,sigB,virR,mprF,dltA,dltB,dltC,dltD,srtB,fbpA and hpt) were detected by PCR.Results showed that in the 91 Lm strains,the detection rate of 23 virulence genes were 100％.The 29 virulence genes of 26 Lm strains were all detected,and 65 Lm strains had different deletion of 6 virulence genes inlD,inlF,inlG,inlH /C2,inlJ and mpl.The deletion rate of inlG and inlF were 60.44％ and 54.95％,respectively,following by mpl gene,with a deletion rate of 19.78％.According to the absence of virulence genes,91 strains could be divided into 10 subtypes,and the dominant virulence subtypes was type Ⅰ with all 23 virulence genes.The deletion rate of virulent genes in Shijiazhuang was higher than that in northern Hebei.It is suggested that the rate of virulence gene of food-borne Lm in Hebei Province is high,and the virulence gene deletion patterns has diversity and regional differences.

OBJECTIVETo explore the effect and the mechanism of yifei jianpi recipe (YFJPR) on patients with chronic obstructive pulmonary disease (COPD).

METHODSForty patients with COPD in stable phase were randomly divided into two groups, the treated group and the control group. Indexes including the total and differential count of inflammatory cell in sputum, levels of interleukin-8 (IL-8) and tumor necrosis factor-alpha (TNF-alpha), as well as the percentage of forced expiratory volume in one second in its predicted value (FEV1%) and ratio of FEV1/forced vital capacity (FVC) in patients were measured before and after treatment, and compared with those in 20 healthy subjects.

RESULTSAll the indexes measured in patients before and after treatment were significantly different from those in healthy subjects (P < 0.01). Differential count of polymorphonuclear neutrophil (PMN) and levels of IL-8 and TNF-alpha in sputum in the treated group significantly decreased after treatment (P < 0.01), while the non-PMN differential count and levels of FEV1% and FEV1/FVC significantly increased (P < 0.01). But in the control group, changes only showed in increasing of FEV1% and FEV1/FVC (P < 0.05 or P < 0.01). And the effects in the treated group were better than those in the control group (P < 0.01).

CONCLUSIONYFJPR can play a therapeutic role on patients with COPD by way of reducing the airway inflammatory reaction.

Adult ; Aged ; Drugs, Chinese Herbal ; therapeutic use ; Female ; Humans ; Interleukin-8 ; metabolism ; Leukocyte Count ; Male ; Middle Aged ; Neutrophils ; pathology ; Phytotherapy ; Pulmonary Disease, Chronic Obstructive ; drug therapy ; metabolism ; pathology ; Respiratory Function Tests ; Sputum ; cytology ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objectives: To observe the effect of activated G-protein coupled estrogen receptor 1 (GPER1) on Angiotensin II (AngII)-induced hypertrophy of cultured neonatal rat cardiomyocytes and explore related mechanisms. Methods: Primary cardiomyocytes derived from 2-to 3-day-old neonatal rats were cultured in vitro. Tandem mass tags (TMT) protein mass spectrometry was used to examine protein expressions; relevant bioinformatics analysis was performed to screen the possible regulatory mechanisms. Cardiomyocytes were divided into 6 groups: (1)Blank control group, (2) AngII group, (3)AngII+G1 (GPER1 activator) group, (4)AngII+G1+G15 (GPER1 inhibitor) group, (5)AngII+G1+U0126 (extracellular ERK inhibitor) group and (6)AngII+G1+MK2206 (AKT inhibitor) group (n=3 for each group). Cardiomyocytes GPER1 expressions, mRNA levels of atrial natriuretic peptide (ANP) and B-type natriuretic peptide (BNP), protein levels of ERK, AKT with their interactions, autophagy-related proteins LC3II and LC3I were compared among different groups;impact of GPER1 on cardiomyocytes apoptosis was detected by flowcytometry. Results: AngII induced cardiomyocytes hypertrophy and upregulation of ANP and BNP mRNA levels in a dose-dependent manner (P<0.05). GPER1 expression could be detected on cardiomyocytes by Immunofluorescence technique. qRT-PCR results showed that GPER1 was activated by the specific activator G1 and mRNA expressions of ANP and BNP were inhibited in a dose-dependent manner by the specific activator G1 (P<0.05); mRNA levels of ANP and BNP were re-elevated in AngII+G1+G15 group (P<0.05). Western blotting results showed that protein expression of p-ERK and p-AKT was significantly higher in AngII group and AngII+G1 group than in blank control group (P<0.05), significantly reduced in AngII+G1+G15 group compared with AngII+G1 group (P<0.05); decreased expressions of p-ERK, p-AKT and mRNA levels of ANP,BNP were also detected in AngII+G1+MK2206 group (P<0.05). G1 induced protein expression was similar between AngII+G1 group and AngII+G1+U0126 group. Flowcytometry results indicated that cardiomyocytes apoptosis was similar between AngII+G1 group and AngII group (P>0.05). Ratio of LC3II/LC3I was significantly higher and autophagy levels were significantly enhanced in AngII group than in blank control group (P<0.01), these changes could be significantly reversed in AngII +G1 group (P<0.01 vs. AngII). Conclusions: Activation of GPER1 could inhibit neonatal cardiomyocytes hypertrophy, this effect might be related to AKT and ERK signaling pathway and cell autophagy.

OBJECTIVEAllogeneic bone marrow transplantation has been established as a standard method for the treatment of a range of malignant and non-malignant hematologic diseases in children. Unfortunately, fewer than 30% of patients have a human leukocyte antigen (HLA)-matched sibling. Advances in our understanding of the HLA system and the development of large international donor registries encourage the increasing use of unrelated donors as an alternative source of stem cells. The purpose of this study was to evaluate the clinical efficacy and safety of unrelated donor allogeneic bone marrow transplantation (URD-BMT) for the treatment of childhood leukemia.

METHODSSix patients with leukemia received URD-BMT. Two of them suffered from chronic myeloid leukemia (CML), 3 suffered from acute lymphocytic leukemia (ALL) and 1 suffered from acute promyelocytic leukemia (APL) (CR2). All cases were facilitated by Tzu Chi Marrow Donor Registry (TCTMDR). The high resolution DNA test for classIand II was carried out in HLA typing of all donor-receiver pairs. HLA allele matched in three cases, mismatched with one locus in two cases and with two loci in one case. All patients were prepared with cyclophosphamide (CY) 60 mg/kg/day for 2 days (total dose 120 mg/kg) and busulfan (Bu) 1 mg/kg x 4/day for 4 days (total dose 16 mg/kg). Mycophenolate mofetil (MMF), CsA and MTX were given to prevent acute graft-versus-host-disease (aGVHD). CsA of 3 mg/kg/d was continuously given by i.v. infusion, and then 6mg/kg/d by oral. The blood CsA concentration was 200 - 300 ng/ml. MTX was given at the dosage of 15 mg/m(2) on d 1 and 10 mg/m(2) on d 3, 6,9 or 11. MMF was given at the dosage of 0.25 - 0.5 g/d from day 0 to day 120. Prostaglandin E1 was given to prevent the hepatic veno-occlusive disease (VOD), Ganciclovir was used to prevent CMV infection until the CMV antigenemia became negative.

RESULTSAnalysis of DNA short tandem repeats showed total engraftment of donor marrow after transplantation in all cases. The median time when granulocyte exceeded 0.5 x 10(9)/L was 14.5 (13 - 18) days, platelets exceeded 20 x 10(9)/L was 16 (14 - 23) days. The acute GVHD grade II-IV occurred in 2 of 6 (33.3%) patients. There were 3 cases with chronic GVHD and none of them developed with the extensive chronic GVHD. All patients were alive in disease-free situation now with median follow-up 412 (187 - 1338) days.

CONCLUSIONURD-BMT is an effective method for the treatment of childhood leukemia.

Bone Marrow Transplantation ; Child ; Humans ; Immunosuppressive Agents ; therapeutic use ; Leukemia ; therapy ; Tissue Donors ; Transplantation, Homologous ; Treatment Outcome