BACKGROUNDWe investigated the pathogenesis of atherosclerosis in diabetes, and detected the expression of scavenger receptor CD36 in monocytes in patients with type 2 diabetes.

METHODSAccording to the criteria by WHO, diabetic patients were classified into two groups: well controlled diabetic patients (WCP) and poorly controlled diabetic patients (PCP). The expression of CD36 protein and mRNA were evaluated by flow cytometry and reversal transcription polymerase chain reaction (RT-PCR). Plasma levels of accumulation of oxidized LDL (oxLDL) were directly measured by sandwich enzyme-linked immunosorbent assay (ELISA) method.

RESULTSFlow cytometry and RT-PCR showed that the mean fluorescence intensity (MFI) of CD36 in monocyte and CD36 mRNA were significantly higher in the PCP and WCP in comparison with healthy controls (P<0.01). CD36 MFI and mRNA in the PCP were increased by 78% and 36% compared to the WCP. In both groups, CD36 MFI and mRNA were significantly higher in patients with diabetic atherosclerosis in comparison with those without diabetic atherosclerosis (P<0.05). No significant difference was found in CD14 expression between the groups (P>0.05). The concentrations of plasma oxLDL were higher in the PCP group compared to WCP and control group (P<0.05), whereas oxLDL average values did not differ significantly between WCP and control groups (P>0.05). In the WCP and PCP groups, oxLDL levels were higher in patients with diabetic atherosclerosis than those without diabetic atherosclerosis (P<0.05).

CONCLUSIONSThe increased expression of scavenger receptor CD36 may be one of the mechanism of accelerated atherosclerosis in diabetic. The poorly controlled diabetes patients are at higher risk for the vascular complications than the well controlled diabetic patients.

Adult ; Aged ; Atherosclerosis ; etiology ; CD36 Antigens ; genetics ; Diabetes Mellitus, Type 2 ; complications ; Female ; Fluorescent Antibody Technique ; Humans ; Lipoproteins, LDL ; analysis ; Male ; Middle Aged ; Monocytes ; metabolism ; RNA, Messenger ; analysis ; Regression Analysis

OBJECTIVETo explore relationship between HBV DNA level and peripheral blood follicular helper T lymphocyte (Tfh) in patients with chronic hepatitis B (CHB) and its significance.

METHODSHBV DNA levels of 179 cases of CHB patients with positive HBV DNA, positive HBeAg and positive human leukocyte antigen(HLA)-A2 were tested with real time fluorescent quantitative PCR. Tfh and HBV specific CTL were tested with flow cytometry. IL-21 was also tested. 179 cases of CHB patients were divided into group A and group B based on HBV DNA levels, 86 cases in group A, HBV DNA levels were 10(4)-10(5) copies/ml, 93 cases in group B, HBV DNA levels were 10(6)-10(7) copies/ml. Above testing indexes of the two groups were compared.

RESULTSHBV DNA levels of group A were (4.85 +/- 0.37) log10 copies/ml, HBV DNA levels of group B were (6.83 +/- 0.31 ) log10 copies/ml, t = 27.31, P < 0. 001; Tfh of group A was (5.96 +/- 1.59)%, higher than that of group B (3.71 +/- 2.15)%, t = 4.92, P < 0.01; IL-21 of group A was (42.61 +/- 15.11)ng/L, higher than that of group B (14.91 +/- 3.15) ng/L, t = 8.62, P < 0.01; HBV specific CTL of group A was (0.36 +/- 0.08)%, higher than that of group B (0.18 +/- 0.06)%, t = 19.99, P < 0.001.

CONCLUSIONSerum HBV DNA level of CHB patients is related to the level of peripheral blood Tfh level: patients with low HBV DNA level have high Tfh level, high IL-21 level and high HBV specific CTL level. Patients with high HBV DNA level have low Tfh level, low IL-21 level and low HBV specific CTL level. The mechanism of baseline HBV DNA level affecting anti-viral therapy may be related to Tfh level.

Adult ; CD4 Lymphocyte Count ; DNA, Viral ; blood ; genetics ; Female ; HLA-A2 Antigen ; immunology ; Hepatitis B virus ; genetics ; isolation & purification ; Hepatitis B, Chronic ; blood ; immunology ; virology ; Humans ; Interleukins ; immunology ; Male ; T-Lymphocytes, Helper-Inducer ; cytology

Abstract: This study is to improve the affinity of scFv-AK404R against VEGFR2. The secondary mutational library was constructed by hydrophilic shuffling in CDR3 region of the heavy chain. VEGFR2-specific screening was performed by phage display technology and the protein of mutants was expressed in periplasm of E.coli HB2151 and purified by affinity chromatography. The affinity constant of scFvs was measured by competitive ELISA, and the structure of scFvs was analyzed by bioinformatics. The result showed that a library with 6.4x10(5) scFv members was established by electro-transformation. Two mutated clones with high absorbance value were isolated after screening. After purification by affinity chromatography, electrophoretically pure scFv proteins were obtained. The competitive ELISA showed that the affinities of WZ01 and WZ02 were three times higher than that of the parental AK404R, and bioinformatics analysis showed that the enlarged contact surface and fitted closely with KDR3 surface may be the reasons for improved affinity. These results suggest that introducing hydrophilic amino acids to the heavy chain CDR3 region is an effective approach to improve the affinity of scFv.
Amino Acid Sequence ; Antibody Affinity ; Chromatography, Affinity ; Computational Biology ; Enzyme-Linked Immunosorbent Assay ; Escherichia coli ; metabolism ; Hydrophobic and Hydrophilic Interactions ; Peptide Library ; Single-Chain Antibodies ; genetics ; immunology ; isolation & purification ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; immunology ; metabolism

OBJECTIVEThe anemia of chronic disease (ACD) is usually defined as mild to moderate anemia occurring during the chronic infection, inflammation, neoplasm or trauma. It is the most common anemia among in-hospital adults. The insufficient endogenous erythropoietin (EPO) production is probably one of the pathogenic mechanisms of ACD. Inflammatory cytokines play an important role in the ACD pathogenesis. But nowadays there are few published papers on the childhood ACD in the world. This study aimed to detect the EPO levels in children's ACD, to explore the relationship between EPO and tumor necrosis factor alpha (TNF alpha) and interleukin-6 (IL-6) and, to evaluate the effect of recombinant human TNF alpha (rhTNF-alpha) on EPO gene expression.

METHODSSixty children were divided into ACD group (20 children), non-anemia (NA) group (19 children) and iron deficiency anemia (IDA) group (21 children) according to clinical diagnosis. Serum TNF alpha and IL-6 levels were detected with ELISA method. The EPO level was detected by chemical immulite method. The effect of rhTNF alpha on the expression of EPO gene was studied by culturing Hep G2 cell line and RT-PCR method.

RESULTSSerum EPO levels were different among the 3 groups (F = 44.68, P < 0.01). Serum EPO levels in ACD group were higher than those in NA group, while the hemoglobin levels were similar between the two groups. Serum EPO levels in ACD patients were lower than those in IDA patients. Serum TNF alpha levels were different among the 3 groups (F = 25.15, P < 0.01), and serum IL-6 levels were also different among the 3 groups (F = 13.16, P < 0.01). Serum TNF alpha and IL-6 levels in ACD group were higher than those in NA group. In ACD group, serum levels of both TNF alpha and IL-6 were not correlated to the serum level of EPO (r = -0.35, P > 0.05 and r = -0.05, P > 0.05, respectively). In vitro, rhTNF alpha inhibited the expression of EPO mRNA in hypoxia, and the inhibitory effects became stronger with the increase of rhTNF alpha (F = 64.20, P < 0.01).

CONCLUSIONEPO levels increased incompensatively in ACD children, which may be a cause of ACD. TNF alpha may cause anemia by inhibiting EPO production.

Anemia ; blood ; genetics ; Cell Line, Tumor ; drug effects ; metabolism ; Child ; Child, Preschool ; Chronic Disease ; Erythropoietin ; blood ; genetics ; Gene Expression ; drug effects ; Humans ; Interleukin-6 ; blood ; genetics ; RNA, Messenger ; genetics ; metabolism ; Recombinant Proteins ; pharmacology ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Necrosis Factor-alpha ; genetics ; metabolism ; pharmacology

OBJECTIVETo compare the difference of HBV DNA levels and HBV genotypes between the patients with primary hepatocellular carcinoma (HCC) and liver cirrhosis who infected with hepatitis B virus.

METHODSTotal 430 patients with hepatitis B were enrolled and further divided into the HCC group (210 cases) and liver cirrhosis group (HBV LC, 220 cases). The levels of HBV DNA and HBV genotypes were detected in all of the serum samples from the two groups, and the differences in the genotypes and virological markers between HCC patients and HBV LC patients were further analyzed.

RESULTSThe positive rates of HBV DNA of HCC patients and HBV LC patients were 84.3% (177/210) and 94. 5% (208/220), respectively. The mean values of serum HBV DNA in HCC patients and HBV-LC patients were (5.06 +/- 1.01) log10 cps/ml and (5.36 +/- 1.13) log10 cps/ml, respectively. The positive rates of HBV DNA and the mean values of serum HBV DNA were higher in HBV-LC patients than those in HCC patients (P < 0.01). Furthermore, the main genotype was C in both groups and the distribution of genotype C and genotype B had no statistical difference.

CONCLUSIONSMainly presented as a C genotype in both groups, the total levels of serum HBV DNA in HCC patients were lower than those in HBV-LC patients.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Hepatocellular ; blood ; virology ; DNA, Viral ; blood ; genetics ; Female ; Genotype ; Hepatitis B ; blood ; virology ; Hepatitis B virus ; genetics ; isolation & purification ; Humans ; Liver Cirrhosis ; virology ; Liver Neoplasms ; blood ; virology ; Male ; Middle Aged

Development of the disease is the result of several factors involved in biological network changes. The nature of drug intervention is to regulate these pathological changes to the normal range. Advantages of traditional Chinese medicine (TCM) are to integrally and systematically regulate this biological networks and systematic pathology through multi-targets, multi-levels, multi-channels. Structural components TCM provides the controlled and precise basis "substance" for this regulation and also to clarify the "truth" of the nature of the regulation by the network pharmacology. Network pharmacology provides new strategy for the research on mechanism of structural components TCM. This study not only reflects the overall characteristics of the development of the disease, but also fully embodies the essence of TCM for preventing and treating diseases through changing traditional model on "one drug, one gene, one disease". This paper explores systematically the integration essence, features and research strategies of structural components TCM and the network pharmacology, understand the interaction of structural components TCM and body from the perspective of the overall concept of improving or restoring the balance of.biological networks. It is effective measure to reveal the structure of a multi-component for regulating biological networks mechanisms, and also provide new ideas and methods for further scientific research and innovation of structural component TCM.
Drug Interactions ; Drug Therapy ; Drugs, Chinese Herbal ; chemistry ; pharmacology ; Gene Regulatory Networks ; drug effects ; Humans ; Medicine, Chinese Traditional

BACKGROUND/AIMS: There has been debate on whether a sodium-restricted diet (SRD) should be used in cirrhotic patients with ascites in China in recent years. The purpose of this study was to compare the effect of sodium-restricted and unrestricted diets on plasma renin activity (PRA), renal blood flow (RBF) and ascites in patients with liver cirrhosis. METHODS: Two hundred cirrhotic patients with ascites were randomly divided into two groups (98 cases in the sodium-unrestricted diet [SUD] group and 102 cases in the SRD group); 95 patients (96.94%) in the SUD group and 97 patients (95.1%) in the SRD group had post-hepatitis B cirrhosis. RESULTS: Blood sodium and RBF were higher in SUD group than in SRD group (p<0.001), while PRA were significantly lower in SUD group than the SRD group 10 days after treatment (p<0.001). Renal impairment caused by low blood sodium was higher in SRD group than in SUD group (p<0.01). Ascites disappeared in higher proportion of patients in SUD group than in SRD group (p<0.001). CONCLUSIONS: SUD can increase the level of blood sodium and RBF, and be beneficial to diuresis and ascite reduction and disappearance.
Ascites ; China ; Diet ; Diet, Sodium-Restricted ; Diuresis ; Humans ; Liver ; Liver Cirrhosis ; Plasma ; Renal Circulation ; Renin ; Sodium

OBJECTIVETo explore the association of serum HBV DNA level with HBV-specific and nonspecific cytotoxic T lymphocytes (CTL) in patients with HBV-induced hepatic cirrhosis.

METHODS120 patients with HBV-induced hepatic cirrhosis who were positive for HBV DNA, HBeAg and human leucocyte antigen (HLA)-A2 were enrolled in this study. The level of HBV DNA was determined by real time fluorescence quantitative polymerase chain reaction (PCR). HBV-specific and nonspecific CTL were detected by flow cytometry. Liver function tests were done in the 120 patients. The 120 patients were divided into group A and B based on their HBV DNA levels. In group A, there were 68 patients with HBV DNA level of 3-4 log10 copy/ml, and in group B, 52 with 5-6 log10 copy/ml. HBV-specific and nonspecific CTL and liver function were compared between the two groups.

RESULTSHBV DNA levels were (3.68 +/- 0.19) and (5.97 +/- 0.32) log10 copy/ml in Group A and B respectively with P < 0.001. HBV-specific CTL was higher in group A (0.33% +/- 0.04%) than in group B (0.11% +/- 0.01%) with P < 0.001. HBV-nonspecific CTL were (11.99% +/- 1.51% ) and (11.91% +/- 1.61%) in group A and B respectively with P > 0. 05.

CONCLUSIONThe level of serum HBV DNA is related to the levels of HBV-specific CTL in patients with HBV-induced hepatic cirrhosis. Patients with higher HBV DNA had lower levels of HBV-specific CTL, and the damage to liver function was severe because of higher levels of HBV DNA. Patients with lower HBV DNA had higher levels of HBV-specific CTL which predict good anti-viral effect.

Adult ; Case-Control Studies ; DNA, Viral ; blood ; Female ; HLA-A2 Antigen ; genetics ; Hepatitis B e Antigens ; blood ; Hepatitis B virus ; genetics ; immunology ; physiology ; Humans ; Liver Cirrhosis ; blood ; immunology ; virology ; Liver Function Tests ; Male ; Middle Aged ; T-Lymphocytes, Cytotoxic ; immunology ; Young Adult

OBJECTIVETo explore influence of sodium restricted diet and non-sodium restricted diet on plasma rennin (PRA), angiotensin II (All), ALD, renal blood flow (RBF) and subside of ascites in patients with cirrhotic ascites.

METHODSEighty cases of hepatitis B with cirrhotic ascites were randomly divided into sodium restricted diet group and non-sodium restricted diet group. 39 cases were in non-sodium restricted diet group, taking sodium chloride 6500-8000 mg daily; 41 cases were in sodium restricted diet group, taking sodium chloride 5000 mg daily. Both groups received diuretics furosemide and spironolactone. Blood sodium, urine sodium, PRA, AII, ALD, RBF ascites subsiding were compared after treatment.

RESULTSIn non-sodium restricted diet group, blood sodium and urine sodium increased 10 days after treatment compared with those before treatment, and compared with those of sodium restricted diet group 10 days after treatment, P <0. 01. RBF increased compared with that before treatment, and compared with that of sodium restricted diet group 10 days after treatment, P < 0. 01. Renal damage induced by low blood sodium after treatment was less in non-sodium restricted diet group than that in sodium restricted diet group, P <0. 05. Ascites disappearance upon discharge was more in sodium restricted diet group than that in non-sodium restricted diet group, P <0. 01. Time of ascites disappearance was shorter in non-sodium restricted diet group than that in sodium restricted diet group, P < 0. 01.

CONCLUSIONCompared with sodium restricted diet, while using diuretics of both groups, non-sodium restricted diet can increase level of blood sodium, thus increasing excretion of urine sodium and diuretic effect. It can also decrease levels of PRA, AII and ALD, increase renal blood flow and prevent renal damage induced by low blood sodium and facilitate subsiding of ascites.

Ascites ; blood ; diet therapy ; physiopathology ; urine ; Chymosin ; blood ; Diet, Sodium-Restricted ; methods ; Diuretics ; administration & dosage ; Female ; Furosemide ; administration & dosage ; Hepatitis B ; blood ; diet therapy ; physiopathology ; urine ; Humans ; Liver Cirrhosis ; blood ; diet therapy ; physiopathology ; urine ; Male ; Middle Aged ; Renal Circulation ; drug effects ; Sodium ; blood ; urine ; Sodium, Dietary ; administration & dosage ; Spironolactone ; administration & dosage

OBJECTIVETo study the methylation status of p73 gene promoter in patients with myelodysplastic syndrome (MDS) and explore its significance with clinical prognosis.

METHODSMethylation of p73 promoter was detected in bone marrow cells from 135 MDS patients and 13 healthy controls by methylation-specific PCR (MSP). The results of MSP were confirmed by bisulfite sequencing. The expression of p73 mRNA was detected by real-time quantitative PCR. Primary bone marrow cells from MDS patients were treated with decitabine, the changes of p73 methylation status and p73 mRNA expression were measured. The role of p73 methylation in the prognosis of MDS and the correlated clinical data were explored.

RESULTSp73 hypermethylation was present in 37.04% of MDS cases and patients with high risk MDS (RAEB-1 and RAEB-2) exhibited a significantly higher frequency of p73 methylation than that of low risk MDS (58.8% vs 29.7%, P = 0.002). The expression of p73 mRNA in the methylated group was decreased compared to that of the unmethylated group (P = 0.032). Decitabine treatment decreased the level of p73 methylation and increased the level of p73 transcripts. Patients with p73 methylation progressed rapidly to AML (P < 0.001) and had shorter survival (P = 0.002) than those who did not have p73 methylation. In the multivariate Cox regression model, BM blast and p73 methylation status emerged as independent prognostic factor for overall survival and leukemia free survival.

CONCLUSIONp73 gene methylation is common in patients with MDS and may indicate poor prognosis. p73 may be a therapeutic target in MDS.

Aged ; Case-Control Studies ; DNA Methylation ; DNA-Binding Proteins ; genetics ; Female ; Humans ; Male ; Middle Aged ; Myelodysplastic Syndromes ; genetics ; Nuclear Proteins ; genetics ; Prognosis ; Promoter Regions, Genetic ; Tumor Protein p73 ; Tumor Suppressor Proteins ; genetics