Genes expression profiling of peripheral blood cells of patients with hepatocellular carcinoma.

HCC (hepatocellular carcinoma) is often diagnosed at an advanced stage with poor prognosis. Peripheral blood may be useful in cancer classification, and therefore we investigated the gene expression found by Affymetrix HG-U133 Plus2.0 microarray, with samples from nine HCC patients and five healthy NC (normal controls). A total of 726 probe sets showed significant differences based on the criteria of P<0.05 and absolute fold change >2. The genes were related to many biological functions, including immune response, transcription regulation and metabolism processes. Ten genes [IL-8 (interleukin 8), GOS2 (G0 /G1 switch gene 2), CXCR4 (CXC chemokine receptor 4), FOS, RPS24 (40S ribosomal protein S24), HAP90AA1, PFDN5, RPL27, GZMA and PFN1] showing significant differences were confirmed by real-time PCR in 54 HCC patients and 56 healthy NC. Seven genes [IL-8, GOS2, CXCR4, FOS, RPS24, HSP90AA1 (heat shock protein 90AA1) and PFN1] showed significant difference both in RT-PCR (reverse transcription-PCR) and microarray. Expression of IL-8 and FOS proteins was up-regulated in HCC compared with healthy controls. A gene signature in peripheral blood which can distinguish HCC patients and healthy controls may have been identified.

Increased thyroid function and elevated thyroid autoantibodies in pediatric patients with moyamoya disease: a case-control study.

BACKGROUND AND PURPOSE: The purpose of this study was to investigate whether thyroid function and thyroid autoantibodies were associated with the risk of moyamoya disease in pediatric subjects. METHODS: Thyroid function and thyroid autoantibodies were evaluated in patients with moyamoya disease and control subjects, and their associations with moyamoya disease were estimated using multivariate analysis. RESULTS: We included 114 pediatric patients and 114 healthy control subjects. The patients displayed higher prevalence of increased thyroid function and elevated thyroid autoantibodies in comparison with control subjects. These remained significant after multivariate adjustment; the ORs (95% CI) for increased thyroid function and evaluated thyroid autoantibodies were evaluated as 12.47 (1.55 to 100.51) and 4.33 (1.29 to 14.59), respectively. CONCLUSIONS: Increased thyroid function and elevated thyroid autoantibodies are associated with moyamoya disease and therefore monitoring of thyroid function and thyroid autoantibodies in patients with moyamoya disease is suggested, which might help to guide subsequent clinical management.

Association of a functional polymorphism in the MMP-3 gene with Moyamoya Disease in the Chinese Han population.

BACKGROUND: Moyamoya disease (MMD) is an uncommon cerebrovascular disease characterized by progressive stenosis of the intracranial internal carotid arteries and their proximal branches. The important role of genetic factors in the etiology and pathogenesis of MMD is being increasingly recognized. The study was designed to examine the association of single nucleotide polymorphisms in matrix metalloproteinase (MMP) and tissue inhibitors of metalloproteinase (TIMP) genes with MMD occurrence. METHODS: A case-control study was performed. Five functional promoter polymorphisms in the MMP-2, MMP-3, MMP-9 and MMP-13 genes and a potentially functional promoter polymorphism in the TIMP-2 gene were determined by polymerase chain reaction-restriction fragment length polymorphism. Their associations with MMD were analyzed by multivariate logistic regression. RESULTS: In total, 208 definite patients with MMD (including 31 familial MMD, FMMD, patients) and 224 healthy subjects were recruited. The frequency of the MMP-3 5A/6A and 5A/5A genotypes was significantly lower in MMD patients (OR = 0.57, 95% CI 0.38-0.86, p(corr) = 0.042) compared with healthy controls in a dominant genetic model. Significant differences of the MMP-3 5A/6A polymorphism were also detected between FMMD patients and controls both in the dominant genetic model (OR = 0.23, 95% CI 0.08-0.68, p(corr) = 0.048) and the additive genetic model (OR = 0.24, 95% CI 0.08-0.69, p(corr) = 0.048). CONCLUSION: The functional polymorphism in the MMP-3 promoter might be associated with susceptibility to both MMD and FMMD in the Chinese Han population. The findings need to be validated in further studies including more subjects from different populations.

[Comparison of serum biochemical features between SARS and other viral pneumonias].

OBJECTIVE: To identify blood chemistry changes in coronavirus of severe acute respiratory syndrome (SARS). METHODS: Biochemical changes in SARS patients were summarized and compared with other viral pneumonias. Serum total protein (TP), albumin (Alb), total cholesterol (TC), triglyceride (TG), calcium (Ca), ferrum (Fe), lactate dehydrogenase (LDH), creatine kinase (CK), alanine aminotransferase (ALT) and aspartate aminotransferase (AST) between SARS and other viral pneumonias were examined by Roche Diagnostics assay, HITACHI7600 automatic analyzer. Clinically confirmed SARS patients, patients with other viral pneumonias, and healthy controls were included in the study. RESULTS: Compared with healthy person, the levels of serum TC, Fe, Ca, Alb were significantly lowered (P<0.05 or P<0.01), while the activity of LDH, CK, ALT, AST were elevated, the increase of CK and the decrease of Fe were the most significant (P<0.05 or P<0.01), the changes of TP and TG were not obvious. In the other viral pneumonias patients, ALT, AST, LDH were elevated slightly than those of healthy person, while Fe, Ca, Alb, TC, CK were a little reduced, but there was no significant difference between the two groups. In convalescent stage, all the tests were returned to normal ranges except ALT, AST were still elevated in SARS patients. CONCLUSION: The changes in serum biochemistry are more marked in SARS patients compared with patients suffering from other viral pneumonias, the decrease of Fe as well as the inhibition of TC may be caused by the treatment of anti-virus.

[Experimental study of the effects of mild hypothermia on the acute myocardial infarction size in the rabbits with coronary artery reperfusion].

OBJECTIVE: To study the effect of mild hypothermia on the acute myocardial infarction size in the rabbits with coronary artery reperfusion. METHODS: fourteen rabbits were randomly divided into two groups: mild hypothermia group and control group. Each group underwent 45 minutes of left anterior descending coronary artery occlusion and 2 hours of reperfusion. Core temperatures were measured with thermistor. The mild hypothermia group received ice cooling around the body and the core temperature was dropped to 32-35 centigrade after occluded for 30 minutes, while the control group's body temperature were kept above 38 centigrade. The myocardial area at risk and the infarct area were determined with Evan's blue dye and triphenyl tetrazolium chloride (TTC). RESULTS: The total elevated amplitude of ST segment in chest leads V1, V3 and V5 in the mild hypothermia group was (25.8+/-8.5) mV, it was lower than that in the control group (37.7+/-6.5) Mv (P=0.021). The changes of serum MB isoenzyme of creatine kinase (CK-MB) activities in mild hypothermia group was (2646.9+/-1227.3) U/L, it was significantly lower than that in the control group (4787.8+/-1934.2) U/L(P=0.045). The weight of infarct myocardium of the mild hypothermia group was (0.23+/-0.05)g, it was lower than that in the control group (0.42+/-0.16)g (P=0.020). Myocardial infarct size as a percentage of the risk zone (0.214+/-0.044 vs. 0.357+/-0.066, P=0.001) and of the left ventricle weight (0.041+/-0.010 vs. 0.071+/-0.027, P=0.029) were smaller than those in the control group. The ratio of the survived myocardial area over the risk zone in the mild hypothermia group was significantly higher than that in the control group (0.786+/-0.044 vs. 0.643+/-0.066, P<0.001). CONCLUSION: Mild hypothermia may reduce infarct size in the rabbits with transient acute myocardial infarction, and increase survived myocardium in the risk zone.

[Studies of bi-derectional modulation effect of kudzuvine root on immunol cells].

OBJECTIVE: To evaluate bi-directional modulation effect of Chinese herbal medicine on immunol cells. METHOD: Two different active portions were isolated from Kudzuvine Root(Radix puerariae), one being the ethanol extraction and another the water extraction. Different concentration of these two different portions was studied by using PMA stimulated lymphocyte or eosinophil initiated chemiluminescence system. RESULT: Water extraction of Kudzuvine Root could enhance chemiluminescence concentration dependently whereas enthanol extraction of Kudzuvine Root inhibited the chemiluminescence significantly. CONCLUSION: The bi-directional regulation effect of Chinese herbal medicine can be found in the same herb, because of its efficacy of different active compounds.

Clinical evaluation of the levels of 12 cytokines in serum/plasma under various storage conditions using evidence biochip arrays.

Cytokines are a group of peptides which form a sophisticated network to modulate multiple cellular events. Within such a network, and through complex feedback mechanisms, cytokine functions are largely interdependent and closely associated with a number of pathological processes. In the present study, the EVIDENCE 180 system was used to study the effects of storage temperature and repeated freeze/thaw cycles on the concentration of 12 cytokines in various sample types. Samples were collected from 9 healthy volunteers and stored by 3 methods: gel, glass and lithium heparin (LH) tubes. Immediately following collection, the concentration of each cytokine in the samples was measured. Cytokine concentrations of the 3 sample types that did not undergo repeated freeze/thaw cycles were compared with those subjected to 1­10 freeze/thaw cycles. In addition, the dynamic changes of 6 sample types which were stored at 4˚C for 6 h to 6 days was analyzed. In addition, the within­ and between­run precision of 12 cytokines on the biochip array was evaluated. Interleukin (IL)­8, vascular endothelial growth factor (VEGF) and epidermal growth factor (EGF) concentrations were lower in plasma compared with serum. Cytokine levels in serum and plasma were affected by several freeze/thaw cycles with IL­1ß, ­4 and ­10 increasing significantly following 1 freeze/thaw cycle and remaining at stable increased levels for the duration of the additional 9 cycles. In separated serum samples in gel and glass tubes stored at 4˚C for 6 days, no difference in concentration of the 12 cytokines was identified. In the other 4 sample types, IL­8, VEGF, tumor necrosis factor α and EGF levels were altered when stored at 4˚C. Results indicate that the EVIDENCE 180 system is stable and plasma was observed as the best sample type to determine concentration of the 12 cytokines using this biochip array. Repeated freeze/thaw cycles and storage at 4˚C was identified to affect the concentration of the 12 cytokines. The current study demonstrates that repeated freeze/thaw cycles of samples must be avoid. In addition, results indicate that plasma or serum must be separated immediately following centrifugation and sample concentration should be measured as soon as possible.

Impact of chronic kidney disease on serum tumor markers concentrations.

BACKGROUND: Serum tumor markers have always been of clinical importance in the diagnosis, monitoring disease progression and therapy efficacy for patients with malignant diseases. However, elevated serum tumor markers are found in some benign conditions, especially in chronic kidney disease (CKD). The elevation of them in CKD might cause confusion and misuse of these tumor markers. We conducted this retrospective study to investigate which of the five widely used tumor markers including carcinoembryonic antigen (CEA), alpha-fetoprotein (AFP), cytokeratin 19 fragment antigen 21-1 (Cyfra21-1), squamous cell carcinoma antigen (SCC) and neuron specific enolase (NSE) are affected markedly by CKD, in order to use them more effectively. METHODS: Serum tumor marker concentrations, biochemical, hematological parameters, and urinalysis were measured in CKD patients and healthy controls. The positive rate and median tumor markers' level in CKD patients and controls, and those in CKD patients stratified by CKD grade were compared using nonparametric rank tests. Correlation analysis of serum tumor markers and other parameters in CKD patients were performed using the Spearman correlation coefficient. Multivariate Logistic regression analysis was used to estimate the important variables that caused elevated serum concentrations of these markers in CKD patients. RESULTS: The overall positive rates and serum concentrations of Cyfra21-1, SCC, CEA in CKD group were significantly higher than those in control group. Positive rate and serum concentrations of those tumor markers increased as kidney function decreased. Both univariate analysis and multivariate regression analysis showed that the elevations of those tumor markers were not only associated with kidney function, but also with nutritional status. CONCLUSIONS: Serum concentrations of Cyfra21-1, SCC, CEA are significantly influenced by kidney function, as well as nutritional status. Therefore, in clinical work, the indices of kidney function and nutritional status could be simultaneously measured to improve interpretation of the results of those tumor marker concentrations.

[Feasibility of peptide mass fingerprinting for differential diagnosis of IgA and non-IgA nephropathy].

OBJECTIVE: To investigate the feasibility of peptide mass fingerprinting for non-invasive differential diagnosis of IgA nephropathy (IgAN) from the non-IgA nephropathy (IgAN).? METHODS: According to the results of renal biopsy, 56 patients were divided into IgAN group (n=28) and non-IgAN group (n=28), and peptide mass fingerprints were acquired from these patients using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). RESULTS: Nine different peptides were identified between IgAN and non-IgAN. The two most distinctive differentially expressed peptides, with peptide peak values of 4476.46 and 1968.10, showed area under curve values of 86.18% and 79.77%. Principal component analysis demonstrated that the accumulated explained variance of the first 8 differential peptides reached 95%, suggesting the feasibility of differential diagnosis of IgAN from non-IgAN. Comparison with the Matrix protein database identified the peptide with a relative molecular mass of 5338.08 as a fragment of mucin 4 inform and the 2082.77 peptide as fragment of α1-II type collagen inform. CONCLUSION: MALDI-TOF MS is feasible for differential diagnosis of IgAN and non-IgAN and also has great potentials in the classification of the subtypes of other systemic diseases.

[The study on metabolic difference of human body affected by active stress and passive stress under special events].

OBJECTIVE: To study the metabolic difference of body influenced by active stress and passive stress under special events. METHODS: To detect serum multiple biochemistry index of 57 earthquake rescue medical team and 13 victims of a natural calamity in Wenchuan earthquake by using Hitachi 7600 automatic analyzer. RESULTS: Stress affected biochemistry index deeply. To compared with rescue medical team, the serum ADA, ALP and TG of victims increased obviously and TP, ALB, MAO, Cr, UA, K, Na, Cl, Ca, ApoA1 and HDL decreased obviously. CONCLUSION: Many biochemistry index have been changed under stress and it relate with stress extent. The human body function status was better in active stress than in passive stress.

Searching for Potential Ovarian Cancer Biomarkers with Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass Spectrometry.

Ovarian cancer is a common gynecological malignant disease, causing more deaths among women .The key objective in the treatment of ovarian cancer is early diagnosis. The objective of our study was to seek new ovarian cancer biomarkers based on a serum protein profile with the aim of discriminating ovarian cancer patients from healthy controls. An MB-WCX kit was used to analyze serum samples obtained from 20 ovarian cancer patients and 20 healthy controls and then we generated MALDI-TOF protein profiles from the analysis. After pre-processing of the spectra, linear analysis with ClinProTools bioinformatics software was used to classify protein profiles and search for prominent peaks that could be used as potential ovarian cancer biomarkers. Using ClinproTools bioinformatics and statistical software, we found 5 prominent expressed proteins in the ovarian cancer and healthy control groups. The mass to charge ratio were 4648.21(m/z), 9294.03(m/z), 3886.1(m/z), 9066.38(m/z) and 4254.71(m/z), respectively, and the former four proteins were expressed higher in the ovarian cancer patients, but the later one was expressed at lower levels in the cancer patients. The sensitivity and specificity were both more than 90%. From our study, we found that MALDI-TOF MS is a high-throughput sample preparation method and is a new potential tool for the diagnosis of human disease, not only to search for new early detection biomarkers in the ovarian cancer patients' serum samples, but also with a potential use for routine clinical work.

[Correlation of polymorphism in IL-6 gene promoter with BMI, inflammatory factors, and pathogenesis and progression of CHD].

This study was aimed to explore the correlation between IL-6 gene promoter polymorphisms and coronary heart disease (CHD) by investigating the polymorphisms (-572G/C, -597G/A) in IL-6 gene promoter area, body mass index (BMI), inflammatory factors and other biochemical parameters in Han nationality of North China. The genotypes of IL-6 gene promoter-572G/C, -597G/A were detected by fluorescent probe hybridization with fluorescent resonance energy transfer and melting curve techniques in 194 CHD patients and 123 healthy people as control. The effects of genotype on plasma lipids, apoproteins, high sensitive C-reactive protein (hsCRP) and BMI were also studied. Logistic regression was performed to observe the risk factors of CHD. The results indicated that genotype of IL-6 gene promoter -597G/A polymorphism in 7 cases were GA and were GG in others, whereas no AA genotype had been found and no associations between polymorphism of IL-6 gene -597G/A, BMI and inflammatory factors were found. No differences had been found between the frequencies of IL-6 gene -572G/C genotypes and alleles in CHD and control group. However, significant difference was found between the G allele carrier (GG+GC) and non-G allele carrier (CC) of CHD and control group (p=0.0425). In the control group, median levels of systolic blood pressure of G allele carrier were significant higher than non-G allele carrier (p=0.02). Among all the subjects, median levels of BMI, hsCRP and systolic blood pressure in the group of G allele carrier were significantly higher than that in the group of non-G allele carrier, p values were 0.026, 0.022, 0.005 respectively. Multivariate logistic regression analysis showed that age, triglyceride, sex, high blood pressure, apoprotein C2, cholesterol and lipoprotein-a) were the risk factors for CHD, and apoprotein A1 was a protective factor. The G allele of IL-6 gene -572G/C has been not found to be a risk factor for CHD. It is concluded IL-6 gene -597G/A polymorphism is not correlated with the susceptibility to CHD; IL-6 gene -572G/C polymorphism may be correlated with the susceptibility to CHD in Han nationality of North China, the mechanisms may be related with the changes of BMI, hsCRP and blood pressure levels resulted from the polymorphism of IL-6 -572G/C.