Reference intervals established using indirect method for serum ferritin assayed on Abbott Architect i2000SR analyzer in Chinese adults.

BACKGROUND: Serum ferritin (SF) test has been widely used in clinical practice. However, its reference intervals (RIs) vary depending on the analytical method and ethnic origin. This study was to establish the RIs using indirect method for SF in Chinese adults. METHODS: SF was assayed on Abbott i2000SR analyzer. The SF test results of all health examinees (8913 males aged 18-93 years and 5397 females aged 18-90 years) between December 2010 and April 2019 were obtained from our laboratory information system. After Box-Cox transformation of raw data and exclusion of outliers, parametric and non-parametric approaches were used to calculate 95% RIs. The correlation between SF levels and ages, and the differences in SF levels between subgroups were also analyzed. RESULTS: SF levels in females were significantly different from those in males (Z = 88.96, Z* = 23.17; Z > Z*) and showed a weak positive correlation with age (r = .466, P < .0001). The RIs based on parametric approach in males were 66.12-561.58 µg/L, whereas in all females were 3.59-269.59 µg/L, females aged <50 years 3.26-148.02 µg/L and those aged ≥50 years 17.28-303.27 µg/L. The RIs based on non-parametric approach in males were 65.00-571.37 µg/L whereas in all females were 4.00-254.00 µg/L, females aged <50 years 4.00-152.00 µg/L and those aged ≥50 years 16.00-304.05 µg/L. CONCLUSIONS: Our indirect RIs for SF were markedly different from the manufacturer's recommended RIs and might be more suitable for Chinese adults, which would be helpful in interpreting laboratory data and clinical decision-making.

Use of gastric-acid suppressants may be a risk factor for enteric peritonitis in patients undergoing peritoneal dialysis: A meta-analysis.

WHAT IS KNOWN AND OBJECTIVE: Mounting evidence suggests that long-term use of gastric-acid suppressants (GASs) may be associated with adverse effects. Whether GAS use increases the risk of enteric peritonitis in patients undergoing peritoneal dialysis (PD) is not known. The aim of this meta-analysis was to evaluate the association between GAS use and enteric peritonitis in PD patients. METHODS: We searched PubMed, Embase and Cochrane Library databases from inception to 23 January 2018 to identify eligible studies. The primary outcome was an association between GAS use and enteric peritonitis in PD patients. RESULTS AND DISCUSSION: Six studies involving 829 people were included in this meta-analysis. Pooled data showed that GAS use in PD patients was associated with an increased risk of enteric peritonitis (odds ratio [OR] = 1.27; 95% confidence interval [CI]: 1.02-1.57, I2  = 48%). Subgroup analyses based on GAS type revealed that histamine-2 receptor antagonists (H2 RAs) might increase the risk of enteric peritonitis in PD patients (OR = 1.40; 95% CI: 1.01-1.93; I2  = 8%), but proton pump inhibitors (PPIs) might not (1.13; 0.72-1.77; 6; 34%). WHAT IS NEW AND CONCLUSION: Gastric-acid suppressants use might be a risk factor for enteric peritonitis in PD patients. In particular, H2 RAs increased the risk of enteric peritonitis, but PPIs did not. Therefore, to prevent enteric peritonitis, H2 RAs should probably be prescribed with caution for PD patients.

Computerized tomography findings in calcified signet-ring gastric cancer receiving chemotherapy: a case report.

BACKGROUND: Calcification in primary gastric cancer is very rare. In this report, we describe the computerized tomography (CT) changes in calcification in a patient with locally advanced signet-ring gastric cancer treated with chemotherapy. CASE PRESENTATION: A 49-year-old man presented with 5 months' history of abdominal pain, anorexia, and rapid weight loss. He had undergone Billroth-II subtotal gastrectomy for a bleeding gastric ulcer 30 years ago. Abdominal CT showed irregular thickening of the gastric wall and miliary calcifications. Histologic examination of specimen obtained by endoscopic biopsy showed poorly differentiated calcified signet-ring gastric cancer. The patient was clinically staged T4N2M0 and treated with docetaxel, cisplatin, and fluorouracil (DCF)/oxaliplatin and S-1 (XLOX)/S-1. After five cycles of chemotherapy, the general condition of the patient improved and tumor markers (CEA, CA125, CA199) decreased. However, follow-up CT scans showed continuing increase in the calcification. CONCLUSIONS: To conclude, in this case report we have described the dynamic changes in calcification in a gastric cancer patient receiving chemotherapy. One explanation for the observed increase in calcifications could be that the ischemic necrosis resulting from chemotherapy creates an alkaline environment, which promotes deposition of calcium salts. Our theory needs to be confirmed with histological evidence from a large series of patients. Nevertheless, we hope that these findings will improve understanding of the mechanism of calcification in gastric cancer.

TALENs-mediated homozygous CCR5Δ32 mutations endow CD4+ U87 cells with resistance against HIV­1 infection.

Since evidence suggests that transplantation of bone marrow stem cells with the C­C chemokine receptor type 5 (CCR5)Δ32/Δ32 genotype may cure patients infected with human immunodeficiency virus (HIV)­1, the present study aimed to reproduce the CCR5Δ32 mutation in cluster of differentiation (CD)4+ U87 cells using genome engineering methods. A modified transcription activator­like effector nucleases (TALENs) technique, combined with homologous recombination for site­specific, size­controlled and homozygous DNA deletions, was used to reproduce the homozygous CCR5Δ32 mutation in CD4+ U87 cells. The results indicated that the frequency of the TALENs­targeted mutation reached 50.4% without any selection, whereas homologous recombination from CCR5 to CCR5Δ32 occurred in 8.8% of targeted cells. Notably, a HIV­1 challenge test demonstrated that CCR5Δ32/Δ32 CD4+ U87 cells were resistant to HIV infection. In conclusion, engineered CCR5Δ32/Δ32 mutations endowed CD4+ U87 cells with resistance against HIV­1 infection; this site­specific, size­controlled and homozygous DNA deletion technique was able to induce precise genomic editing, i.e., the deletion or insertion of a predetermined length of DNA sequence at a specific locus throughout the genome.

Intrabone marrow injection enhances placental mesenchymal stem cellmediated support of hematopoiesis in mice.

BACKGROUND/AIM: In order to determine the synergistic effects of human placental mesenchymal stem cells (PMSCs) on hematopoiesis in vivo, we compared the intrabone marrow injection (IBMI) with the conventional intravenous injection (IVI). MATERIALS AND METHODS: C57BL/6 recipient mice conditioned with lethal doses of irradiation were transplanted with bone marrow mononuclear cells (MNCs) and bone marrow-derived mesenchymal stem cells (BMSCs) from BALB/c mice by IBMI or IVI. NOD/SCID recipient mice conditioned with sublethal doses of irradiation were transplanted with human umbilical cord blood MNCs (UCB-MNCs) and PMSCs by IBMI or IVI. RESULTS: The number of hematopoietic cells was significantly higher in mice transplanted with BMSCs by IBMI than in those transplanted by IVI in a murine transplantation model (BALB/c→C57BL/6). Moreover, the percentage of human hematopoietic cells in the tibiae of the NOD/SCID mice that were transplanted with PMSCs plus UCB-MNCs was higher than that in mice transplanted with UCB-MNCs alone. In addition, in mice that were transplanted with PMSCs, PMSCs injected by IBMI were more efficient than those injected by IVI. CONCLUSION: Our results not only elucidated the role of PMSCs in promoting hematopoiesis, but also revealed the therapeutic potential of the combination of PMSCs and IBMI in transplantation.

Acupuncture for dysmenorrhoea.

BACKGROUND: Primary dysmenorrhoea is the most common form of period pain and affects up to three-quarters of women at some stage of their reproductive life. Primary dysmenorrhoea is pain in the absence of any organic cause and is characterised by cramping pain in the lower abdomen, starting within the first eight to 72 hours of menstruation.This review examines the currently available evidence supporting the use of acupuncture (stimulation of points on the body using needles) and acupressure (stimulation of points on the body using pressure) to treat primary dysmenorrhoea. OBJECTIVES: To determine the effectiveness and safety of acupuncture and acupressure in the treatment of primary dysmenorrhoea when compared with a placebo, no treatment, or conventional medical treatment. SEARCH METHODS: We searched the following databases: the Cochrane Menstrual Disorders and Subfertility Group Trials Register (to September 2015), Cochrane Central Register of Controlled Trials (CENTRAL) (the Cochrane Library), MEDLINE, EMBASE, PsycINFO, CINAHL and Chinese databases including Chinese Biomedical Literature Database (CBM), China National Knowledge Infrastructure (CNKI), VIP database and registers of ongoing trials. SELECTION CRITERIA: We included all published and unpublished randomised controlled trials (RCTs) comparing acupuncture with sham acupuncture or placebo control, usual care, pharmacological treatment or no treatment. We included the following modes of treatment: acupuncture, electro-acupuncture, and acupressure. Participants were women of reproductive age with primary dysmenorrhoea during the majority of the menstrual cycles or for three consecutive menstrual cycles, and moderate to severe symptoms. DATA COLLECTION AND ANALYSIS: We calculated odds ratios (ORs) for dichotomous outcomes and mean differences (MDs) or standardised mean differences (SMDs) for continuous outcomes, with 95% confidence intervals (CIs). We pooled the data where appropriate. Our primary outcomes was pain. Secondary outcomes included menstrual symptoms, quality of life, and adverse effects. MAIN RESULTS: We included 42 RCTs (4640 women). Acupuncture or acupressure was compared with a sham/placebo group, medication, no treatment or other treatment. Many of the continuous data were not suitable for calculation of means, mainly due to evidence of skew.1. Acupuncture studies Acupuncture versus sham or placebo control (6 RCTs)Findings were inconsistent and inconclusive. However, the only study in the review that was at low risk of bias in all domains found no evidence of a difference between the groups at three, six or 12 months. The overall quality of the evidence was low. No studies reported adverse events. Acupuncture versus NSAIDs Seven studies reported visual analogue scale (VAS) pain scores, but were unsuitable for pooling due to extreme heterogeneity (I² = 94%). In all studies the scores were lower in the acupuncture group, with the mean difference varying across studies from 0.64 to 4 points on a VAS 0 - 10 scale (low-quality evidence). Four RCTs reported rates of pain relief, and found a benefit for the acupuncture group (OR 4.99, 95% CI 2.82 to 8.82, 352 women, I² = 0%, low-quality evidence). Adverse events were less common in the acupuncture group (OR 0.10, 95% CI 0.02 to 0.44, 4 RCTs, 239 women, 4 trials, I² = 15%, low-quality evidence). Acupuncture versus no treatment Data were unsuitable for analysis, but pain scores were lower in the acupuncture group in all six studies reporting this outcome. The quality of the evidence was low. No studies reported adverse events.2. Acupressure studiesNo studies of acupressure reported adverse events. Acupressure versus sham or placebo controlData were unsuitable for pooling, but two studies reported a mean benefit of one to three points on a 0 - 10 VAS pain scale. Another four studies reported data unsuitable for analysis: all found that pain scores were lower in the acupuncture group. No studies reported adverse events. The quality of the evidence was low. Acupressure versus NSAIDsOne study reported this outcome, using a 0 - 3 pain scale. The score was higher (indicating more pain) in the acupressure group (MD 0.39 points, 95% CI 0.21 to 0.57, 136 women, very low-quality evidence). Acupressure versus no treatmentThere was no clear evidence of a difference between the groups on a VAS 0 - 10 pain scale (MD -0.96 points, 95% CI -2.54 to 0.62, 2 trials, 140 women, I² = 83%, very low-quality evidence). AUTHORS' CONCLUSIONS: There is insufficient evidence to demonstrate whether or not acupuncture or acupressure are effective in treating primary dysmenorrhoea, and for most comparisons no data were available on adverse events. The quality of the evidence was low or very low for all comparisons. The main limitations were risk of bias, poor reporting, inconsistency and risk of publication bias.

[Validation of GlobalFiler® PCR Amplification Kit and the STR Polymorphism].

OBJECTIVE: To test the technical parameters of GlobalFiler® PCR Amplification Kit for its application to forensic application value and to investigate the genetic polymorphisms. METHODS: The validation was conducted in sensitivity, mixed samples, species specificity, adaptability, survivability, consistency, peak height balance and stability. The amplification and detection of the genomic DNA from 373 unrelated individuals from Beijing Han nationality were extracted by automation workstation. RESULTS: Global-Filer® PCR Amplification Kit was adaptive to some mixed, degraded and inhibited samples. The power of sensitivity and adaptability and peak height balance showed well. The distributions of genotype frequencies for 21 STR loci in the population were all in accordance with Hardy-Weinberg equilibrium (P > 0.05). The PIC value of the 21 STR loci was among 0.536 to 0.940; the H value was among 0.558 to 0.933; the DP value was among 0.783 to 0.992; the PE value was among 0.243 to 0.874. CONCLUSION: GlobalFiler® PCR Amplification Kit is suitable for criminal cases and DNA database in forensic practice. And 21 STR loci in Beijing Han nationality have high polymorphism, which have application value in forensic practice and population genetics.

[Effects of cyclopamine on the proliferation and apoptosis of LNCaP cells and expression of the PCA3 gene in human prostate cancer].

OBJECTIVE: To explore the effects of cyclopamine on the proliferation and apoptosis of LNCaP cells and the expression of the PCA3 gene in human prostate cancer in vitro. METHODS: LNCaP cells were treated with cyclopamine at the concentrations of 1, 5, 10 and 15 micromol/L for 24, 48 and 72 hours. The inhibitory effects of cyclopamine on the proliferation and apoptosis of the LNCaP cells were detected by MTT and flow cytometry respectively, the morphological changes of the cells observed by Hoechst 33258 staining, and the expression of the PCA3 gene determined by real-time fluorescence quantitative reverse transcriptase polymerase chain reaction (FQ-RT-PCR). RESULTS: Compared with the blank control group, cyclopamine significantly inhibited the proliferation of the LNCaP cells at 5, 10 and 15 micromol/L (P <0.01), reaching IC50 at 10 micro mol/L at 48 hours. The apoptosis rates of the LNCaP cells at 24, 48 and 72 hours were 37.21%, 57.38% and 57.98% in the 10 micromol/L group and 21. 16% , 71.31% and 72.90% in the 15 micro.mol/L group, significantly different from those in the control (P <0. 01). The cell apoptosis showed a rising trend with the increase of cyclopamine concentration and acting-time, while the expression of the PCA3 gene was decreasing with the increased concentration of cyclopamine, significantly lower than that of the blank control group (P <0.01) , and extremely low in the 10 micromo/L group CONCLUSION: Cyclopamine intervention at 10 and 15 micromol/L for 48 and 72 hours could significantly inhibit the at all time points. Proliferation and induce the apoptosis of LNCaP cells and reduce the expression level of PCA3.

Adiponectin exerts its negative effect on bone metabolism via OPG/RANKL pathway: an in vivo study.

To explore the effects of adiponectin on the bone metabolism in vivo. Bone mineral density (BMD), bone microstructure, serum adiponectin levels, and biochemical markers of the bone turnover were measured in 12-week-old male Adipo-/- and WT mice. In addition, the osteoclast formation, osteoprotegerin (OPG), and the receptor activator of nuclear factor-κB ligand (RANKL) expression were examined. The serum adiponectin levels were normal in the WT mice while undetectable in the Adipo-/- mice. Compared with the WT mice, the Adipo-/- mice had higher BMD, more trabecular bone, greater bone volume fraction, and trabecular thickness in the left femur. On the contrary, fewer osteoclasts were observed in the Adipo-/- mice when compared with the WT mice. Meanwhile, the Adipo-/- mice had a significantly decreased serum carboxyl-terminal telopeptide of type 1 collagen (CTX)/osteocalcin (OC) ratio. Interestingly, both the adiponectin and RANKL would cause a significant increase of CTX/OC ratio in the co-culture of the CD14+ peripheral blood mononuclear cells and the osteoblasts from Adipo-/- mice. Further, immunohistochemistry assays in tibias and both the RT-PCR and immunoblot analyses in the cultured osteoblasts showed the Adipo-/- mice expressed lower levels of RANKL but higher levels of OPG. Adiponectin had a negative effect on the bone metabolism, and this negative effect might be mediated, at least in part, by the OPG/RANKL pathway.

[Modified TRIzol method for RNA and DNA co-extraction from blood].

OBJECTIVE: To establish a new method for RNA and DNA co-extraction from the same sample by TRIzol reagent. METHODS: After the aqueous phase which contained total RNA was removed by traditional TRIzol method, the values of pH of the interphase phase and organic phase were adjusted. The DNA was precipitated with ethanol and purified with DNA IQ system. The purified DNA was measured in quality and quantity. As the template, it was amplified and typed by PCR-STR. The data was compared with that extracted by traditional TRIzol method. RESULTS: The DNA extracted by this modified method showed a better result of quality and quantity than that by traditional TRIzol method and a good STR typing. CONCLUSION: The modified TRIzol method is advisable and reliable to simultaneously extract both DNA and RNA from the same sample. It could be used for individual identification and paternity testing to satisfy the need of forensic science.

Survivin up-regulates the expression of breast cancer resistance protein (BCRP) through attenuating the suppression of p53 on NF-κB expression in MCF-7/5-FU cells.

Both breast cancer resistance protein (BCRP, ABCG2) and apoptosis-related molecules are involved in the development of multidrug resistance (MDR) in cancer cells. However, the association of BCRP with apoptosis-related molecules in the development of MDR is unknown. In this study, we investigated the changes in expression levels of BCRP, Survivin, p53, Bcl-2, Bcl-xL or Bax in cultured MCF-7 and MCF-7/5-FU cells, and explored whether these changes affected the expressions of BCRP. Our data showed that the protein and mRNA expression levels of BCRP, Survivin and Bcl-2 were significantly higher in MCF-7/5-FU cells than in MCF-7 cells, while p53 expression lower in MCF-7/5-FU cells than in MCF-7 cells. Knockdown of Survivin or Bcl-2 in MCF-7/5-FU cells and overexpression of Survivin in MCF-7 cells revealed that Survivin had significant association with BCRP expression. Luciferase reporter gene assays showed that Survivin up-regulated BCRP expression at transcriptional level and this response was mediated through NF-κB(p50) pathway. However, may be due to the physical interaction between p53 and Survivin, p53 directly affected Survivin-regulated BCRP expressions. Interestingly, we found that Survivin would suppress p53 expression. Furthermore, our data revealed that Survivin itself had no apparent effect on NF-κB(p50) and BCRP expression when knockdown of p53 in MCF-7 cells; whereas p53 exerted significant inhibitory action on these when knockdown of Survivin. In conclusion, through down regulation of p53 expression, Survivin attenuates the suppressing effect of p53 on NF-κB(p50) expression and then enhances BCRP expression. This may represent a novel strategy for reversal of BCRP drug transporter activity to modulate MDR in cancer cells.

[A new method of identifying the peripheral blood and the menstrual blood].

OBJECTIVE: To explore the tissue-specific gene expressions of the peripheral blood and the menstrual blood, and to search some specific factors to establish an effective method for identifying the peripheral blood and the menstrual blood. METHODS: The specific products of the peripheral blood and the menstrual blood were detected by RT-PCR and separated by electrophoretic technology. RESULTS: Beta-spectrin (SPTB) as one specific marker of peripheral blood and 18S rRNA as a kind of the housekeeping gene were expressed in both the peripheral blood and the menstrual blood. However, matrix metalloproteinase 7 (MMP7) as one specific marker of menstrual blood and human beta defensin 1 (HBD1) as one specific marker of vaginal discharge were only found in the menstrual blood. CONCLUSION: There are differences of specific gene expressions between the peripheral blood and the menstrual blood. They could be accurately distinguished from each other by using the combination of fluorescence technology and RT-PCR to detect the specific identification of mRNA.

[Indirect determination of iodine with methyl isobutyl ketone (MIBK) extraction by atomic fluorescence spectrometer].

In this paper, a new method was developed for indirect determination of iodine with methyl isobutyl ketone (MIBK) extraction by atomic fluorescence spectrometer (AFS). Based upon the complex, which can be extacted perfectly by MIBK, formed between I- and Hg2+ in 0.1 mol x L(-1) of nitric acid, indirectly measuring the iodine in organic phase becomes possible. The effects of extraction conditions and other influence factors were investigated. Under the optimized experimental conditions, the results showed that the linearity between its absorbance and the concentration of iodine was 0-12 microg x L(-1), the detection limit is 0.14 microg x L(-1), and the relative standard deviation is 3.3%. The method has been applied to the determination of iodine in eggs, and the recoveries are in the range of 101.8%-110.4%.

[Butanol extraction combined with dilute hydrochloric acid dissolution-atomic fluorescence spectrometric method for indirect determination of molybdenum in Chinese herbal medicine].

A method for indirectly determining the molybdenum in Chinese herbal medicine by butanol extraction and dilute hydrochloric acid dissolution was established for atomic fluorescence spectrometry. The molybdoarsenate heteropoly acid, formed in the presence of As(V) and ammonium molybdate in 0.3 mol x L(-1) sulphuric acid medium, was separated and enriched in the organic solvent, then the evaporation of organic reagent was implemented and the left residue was dissolved in dilute hydrochloric acid in which the arsenic content was determined on behalf of molybdenum. In the optimum experimental conditions, molybdenum content in 0-15 microg x L(-1) range depicts a good linear relationship, the detection limit and relative standard deviation of 0.44 microg x L(-1) and 1.1% were obtained, respectively. Spiked Chinese herbal medicine samples were determined with the proposed method, and recoveries of 95.6%-101.3% were achieved.

Genome-wide association study identifies two new susceptibility loci for atopic dermatitis in the Chinese Han population.

Atopic dermatitis is a chronic, relapsing form of inflammatory skin disorder that is affected by genetic and environmental factors. We performed a genome-wide association study of atopic dermatitis in a Chinese Han population using 1,012 affected individuals (cases) and 1,362 controls followed by a replication study in an additional 3,624 cases and 12,197 controls of Chinese Han ethnicity, as well as 1,806 cases and 3,256 controls from Germany. We identified previously undescribed susceptibility loci at 5q22.1 (TMEM232 and SLC25A46, rs7701890, P(combined) = 3.15 × 10(-9), odds ratio (OR) = 1.24) and 20q13.33 (TNFRSF6B and ZGPAT, rs6010620, P(combined) = 3.0 × 10(-8), OR = 1.17) and replicated another previously reported locus at 1q21.3 (FLG, rs3126085, P(combined) = 5.90 × 10(-12), OR = 0.82) in the Chinese sample. The 20q13.33 locus also showed evidence for association in the German sample (rs6010620, P = 2.87 × 10(-5), OR = 1.25). Our study identifies new genetic susceptibility factors and suggests previously unidentified biological pathways in atopic dermatitis.

[The effect of cotransplantation of human placenta-derived mesenchymal stem cells in combination with human cord blood mononuclear cells on early hematopoietic reconstitution in SCID mice].

AIM: To evaluate the early hematopoietic reconstitution of implanted SCID mice which were transplanted with human umbilical cord blood-derived mononuclear cells (UCB-MNC) and placenta-derived mesenchymal stem cells (PMSC) by intro-bone marrow injection (IBMI). METHODS: The placenta tissues were digested by collagenase IV and cultured with low-glucose DMEM supplemented with b-FGF. The adherent cells were collected and passaged. The phenotypes of the cultured cells were detected by flow cytometry. The osteogenic differentiation and adipogenic differentiation were induced and detected. SCID recipient mice conditioned with sublethal dose irradiation were transplanted with human UCB-MNC and PMSC by IBMI or intravenous injection (IV). Fifty recipient mice were divided into five groups at random: cotransplantation group A (PMSC+UCB-MNC by IBMI), single transplantation group B (UCB-MNC by IBMI), cotransplantation group C (PMSC by IBMI, UCB-MNC by IV), normal saline control group D (normal saline by IBMI), normal control group E (normal saline by IBMI). There were ten recipient mice in every group. On day 14, the bone marrow cells of every recipient mouse were flushed out from the injected tibias and contralateral tibias, respectively. The percentage of human CD34+ and CD45+ hematopoietic cells was analyzed by flow cytometry. RESULTS: PMSC were isolated and expanded from human placenta, which were successfully induced to osteogenic differentiation and adipogenic differentiation. FACS analyses showed that the phenotypes of PMSC were normal. On day 14 after transplantation in SCID mice, the percentages of human CD34+ and CD45+ hematopoietic cells in the tibias of group B were both significantly lower than them in the injected tibias and contralateral tibias of group A. CONCLUSION: Human PMSC could enhance the early engraftment of UCB-MNC in SCID mice.

[The study of the sensitive and specificity on HBV DNA replication by HBV-lP detecting].

OBJECTIVE: To detect HBV-LP and HBV DNA of the patients with hepatitis B, and to study the sensitive and specificity of HBV-LP detecting and its evaluate on for clinical application. METHODS: The ELISA was used for HBV-LP detecting and RT-PCR for HBV DNA detecting. RESULTS: The sensitive and specificity of HBV-LP and HBV DNA were 64.89%, 99.68%, 60.63% and 100% respectively (P > 0.05); +LR, -LR were 202.78, 60.63, 0.3522 and 0.3937, and there were significance between +LR of the detecting (P < 0.005). CONCLUSION: The sensitive and specificity of HBV-LP and HBV DNA detecting are considerable, +LR of HBV-LP are higher comparing HBV DNA. The HBV-LP is better serology index for detecting replication of HBV DNA.

Enhanced proliferation and activation of peripheral blood mononuclear cells in patients with psoriasis vulgaris mediated by streptococcal antigen with bacterial DNA.

Streptococcal infection is believed to have an intimate relationship with psoriasis, although the pathogenic role of streptococcal DNA is not fully understood. To gain a clearer understanding of these dynamics, we investigated the effect of streptococcal DNA on lymphocyte proliferation and activation as well as cytokine secretion in psoriasis. Peripheral blood mononuclear cells (PBMCs) from psoriatic patients had higher proliferative responses upon stimulation by streptococcal antigen (SA) when compared with those from healthy individuals. Strikingly, this enhanced proliferation of PBMCs was attenuated after administration of SA treated with DNase-I. In addition, CD69 expression levels on T cells, including skin-homing lymphocyte cutaneous lymphocyte-associated antigen positive T cells, and IFN-alpha secretion by PBMCs were also attenuated in patients after stimulation with SA without nucleic acid (non-nucleic acid SA, non-NASA) compared with stimulation with untreated SA. However, activation marker CD86 expression levels on B cells as well as the secretion of IFN-gamma and tumor necrosis factor (TNF)-alpha following stimulation with SA or non-NASA were not significantly altered. Interestingly, the attenuated T-cell activation and IFN-alpha secretion in psoriatic patients could be reconstituted when stimulated by non-NASA combined with synthetic CpG-A, but not when combined with synthetic CpG-B. This study demonstrates the integral function of SA, particularly streptococcal DNA, in the pathogenesis of psoriasis.

The up-regulation of monocyte chemoattractant protein-1 (MCP-1) in Ea.hy 926 endothelial cells under long-term low folate stress is mediated by the p38 MAPK pathway.

OBJECTIVE: Monocyte chemoattractant protein-1 (MCP-1), encoded by the CCL2 gene, plays an important role in the initiation and progression of atherosclerosis. Ea.hy 926 endothelial cells grown under low folate conditions (LO cells) synthesize more MCP-1 mRNA and secrete more MCP-1 protein than folate-replete control cells (HI cells). We investigated the mechanisms underlying the modulation of MCP-1 expression by long-term "folate stress". METHODS AND RESULTS: CCL2 transcription, assessed using promoter-reporter assays, is up-regulated in LO cells relative to HI cells, whereas MCP-1 mRNA stability is unchanged. This quantitative transcriptional bias under chronic low folate conditions is not attributable to differences in active NF-kappaB, but is associated with elevated levels of both total p38 and phospho-p38 that are detectable by Western immunoblotting. Transient, acute methotrexate-mediated folate depletion or exposure to high concentrations of homocysteine (Hcy) had no effect on MCP-1 synthesis by Ea.hy 926 cells. The p38 inhibitor SB-203580 abolished the excess MCP-1 production by LO cells. The quantitative transcriptional bias of CCL2 in LO cells was retained following massive induction by TNF-alpha. CONCLUSION: During long-term folate stress, p38 is the primary determinant of CCL2 transcription. Long-term folate insufficiency "primes" Ea.hy 926 endothelial cells to have a quantitatively more vigorous response to cytokine-mediated inflammatory stress.

Genetic and biochemical determinants of serum concentrations of monocyte chemoattractant protein-1, a potential neural tube defect risk factor.

BACKGROUND: Women with the AA genotype at the (-2518)A>G promoter polymorphism of CCL-2, which encodes the potent pro-inflammatory chemokine monocyte chemoattractant protein 1 (MCP-1), may be at increased risk for having offspring affected by spina bifida. As the A allele at this locus has been associated with decreased transcription of MCP-1 mRNA relative to the G allele, the observed genetic association suggests that the risk of spina bifida may be increased in the offspring of women with low MCP-1 levels. The present study was undertaken to identify potential determinants of MCP-1 levels in women of reproductive age. METHODS: A small cohort of Caucasian and African-American women of reproductive age was recruited to participate in an exploratory investigation of the determinants of several disease-related, biochemical phenotypes, including MCP-1. Subjects completed a brief questionnaire and provided a fasting blood sample for biochemical and genetic studies. Potential biochemical, genetic, and lifestyle factors were assessed for their association with MCP-1 levels using linear regression analyses. RESULTS: In this cohort, MCP-1 levels were significantly higher in Caucasians as compared to African-Americans. Further, among women of both races, there was evidence that MCP-1 levels were associated with smoking status, MTHFR 677C>T genotype, and red blood cell tetrahydrofolate levels. CONCLUSIONS: The results of these analyses indicate that, if maternal CCL-2 genotype is related to the risk of spina bifida, this relationship is likely to be more complex than initially hypothesized, perhaps depending upon folate intake, MTHFR 677C>T genotype, the distribution of folate derivatives, and immune/inflammatory activity.