The objective of this study was to observe the expression of hoxc4 and hoxc6 genes in the process of differentiation of hematopoietic stem cell (HSC) to colony forming unit-T Lymphocyte (CFU-TL) in vitro. and to explore the possible mechanism of HCMV-induced maldevelopment of human cord blood CFU-TL on genetic level through effecting the differentiation progress by human cytomegalovirus (HCMV) with and/or all-trans retinoic acid (ATRA), Normal CFU-TL culture was used as blank control. After detection with MTT, mRNA expression levels in the human cord blood CFU-TL hoxc4 and hoxc6 genes following HCMV infection and ATRA treatment were detected by fluorogenic quantitative reserve transcription polymerize chain reaction (FQ-RT-PCR) method. HCMV of 10(6) plaque formation unit (PFU)/ml was diluted to 0.1 ml 10(5) PFU/ml and added into the infected group. The results showed that the expression of hoxc4 and hoxc6 genes in the differentiation process increased slightly on day 3, and were up to the most on day 7 (p < 0.05), while became lower on day 12 respectively in normal group, HCMV group and ATRA group. Compared with the expression of hoxc6, the expression of hoxc4 was obviously higher in each group (p < 0.05). Compared with the expression of hoxc4 and hoxc6 genes in normal group, the expressions of hoxc4 and hoxc6 in ATRA group were up-regulated remarkably (p < 0.05), while the expressions of hoxc4 and hoxc6 in group HCMV were down-regulated (p < 0.05). It is concluded that the regular expression of hoxc4 and hoxc6 genes mRNA appeared in each group. A positive co-relationship exits between hoxc4/hoxc6 genes and lymphocytic progenitor hematopoiesis. Compared with the expression of hoxc6 gene, the expression of hoxc4 gene is obviously higher in each group. HCMV can down-regulate the expression of hoxc4 and hoxc6 genes and lead to suppression effect on cell morphology, which confirms that the normal hematopoietic lineage determination and maturation rely on the stable and consistent expression of homeobox gene. At the same condition, ATRA (6 x 10(-8) mol/L at 60 nmol/ml) can up-regulate hoxc4 and hoxc6 genes expression. ATRA can up-regulate the expression of hoxc4 and hoxc6 genes.
Cell Line ; Cell Proliferation ; Cytomegalovirus ; genetics ; Cytomegalovirus Infections ; genetics ; Homeodomain Proteins ; genetics ; Humans ; Lymphoid Progenitor Cells ; cytology ; Tretinoin ; pharmacology

Objective To construct a recombinant adenovirus carrying UL138 gene, which was re-lated to the latent infection of human cytomegalovirus, and to investigate the effects of UL138 gene on the functions of THP-1 mononuclear cells. Methods The recombinant adenovirus expressing the UL138 gene was packaged. The titer of the recombinant adenovirus was determined by calculating 50% tissue culture in-fective dose ( TCID50 ) . THP-1 mononuclear cells were infected with the recombinant adenovirus at different multiplicity of infection (MOI) and the optimal MOI was determined (100 PFU/cell) by observing the ex-pression of green fluorescent protein ( GFP ) . Changes in the expression of proinflammatory cytokines by THP-1 mononuclear cells that was induced by overexpressed UL138 were analyzed by quantitative PCR. The expression of chemokines and their receptors were measured by quantitative PCR array. Results The re-combinant adenovirus carrying the UL138 gene was successfully constructed with a titer of 1×1011 PFU/ml. The rate of THP-1 mononuclear cells that was infected with the recombinant adenovirus was 60% at the MOI of 1 ∶ 100. Results of the RT-PCR analysis and Western blot assay further confirmed that the recombinant adenovirus could infect THP-1 mononuclear cells successfully and the expression of UL138 protein increased gradually over time. The overexpressed UL138 in THP-1 mononuclear cells significantly inhibited the expres-sion of IL-18, IL-1β, IL-6, IL-8 and TNF-α as indicated by the results of quantitative PCR. Results ob-tained from the quantitative PCR array analysis showed that most of the chemokines and their receptors were downregulated in the transfected THP-1 mononuclear cells except for the chemokines of CCL17, CCL21, CCL2 and XCL2 and the receptors of CCR2, CXCR1, CXCR2, CXCR4 and CX3CR1 which were upregulat-ed. Conclusion We successfully constructed the recombinant adenovirus carrying UL138 gene which could be used to infect THP-1 mononuclear cells. Overexpressed UL138 in THP-1 mononuclear cells significantly affected the functions of THP-1 mononuclear cells.

OBJECTIVETo investigate maternal-fetal transmission at human bocavirus (HBoV).

METHODSIgG antibody to HBoV in serum samples of 316 mothers were determined with ELISA and HBoV DNA was determined with real time PCR in the sera of the mothers and their infants.

RESULTSHBoV-IgG was positive in 40.20 percent (127/316) of the mothers, while it was positive in 29.43 percent (93/316) of the cord blood specimens of the infants. The difference between the two groups was significant (X2=8.12, P less than 0.005); 93 samples of both the mothers and the infants were positive for HBoV-IgG.

CONCLUSIONHBoV-IgG can cross the placenta to the fetuses through placenta. Further study is needed to answer the question whether vertical maternal-fetal transmission occurs.

Adult ; Antibodies, Viral ; blood ; Bocavirus ; DNA, Viral ; blood ; Enzyme-Linked Immunosorbent Assay ; Female ; Humans ; Immunoglobulin G ; blood ; Infant, Newborn ; Infectious Disease Transmission, Vertical ; Parvoviridae Infections ; transmission ; Pregnancy ; Prospective Studies

AIM:To observe the concentration of RBP4 and IL-6 in vitreous of proliferative diabetic retinopathy (PDR). METHODS: A total of 65 patients (66 eyes) were enrolled in Department of Ophthalmology, Renmin Hospital of Wuhan University from February 2017 to July 2017 with the informed consent. The patients were divided into PDR group (23 cases) and NPDR group (16 cases). Twenty- six patients without diabetic mellitus (DM) served as control group. The demography was matched among the groups, but the course of DM, the blood glucose level and the HbA1c level were elevated in the PDR group and the NPDR group (all P<0.05). Vitreous samples were collected during the procedure of vitrectomy. RBP4,IL-6,TNF-α concentrations in vitreous specimens were detected by ELISA. The differences of vitreous RBP4, IL-6 and TNF-α in various groups were statistically analyzed by ANOVA, respectively. The correlations between RBP4 and IL - 6, TNF - α were calculated by Pearson correlation analysis. RESULTS:The concentration of RBP4 in PDR group,the NPDR group and control group were 13.68士2.66, 11 03士1 12,10.45士1.17μ g/Ml, and the concentration of IL-6 were 56.0士10.27, 20.92士5.77, 10.26士1.91pg/Ml. RBP4 and IL- 6 concentrations were elevated in PDR group compared with NPDR group and control group, with significant difference among three groups (F = 12. 135, 161.167; P < 0. 01). IL - 6 concentrations in vitreous increased in the NPDR group in comparison with control group(P<0.05). RBP4 concentrations had no significant difference between the NPDR group and the group(P>0 05). Pearson correlation coefficient was significant positive between RBP4 concentration and IL - 6 concentration(r=0.606,P=0.001). CONCLUSION: RBP4 is probability involved in the inflammation pathogenesis of PDR. These results indicate that RBP4 could be a new target for the diagnosis and treatment of PDR.

Objective: To assess the diagnostic values of ¹¹C-methionine (MET) PET/CT semiquantitative parameters for detecting recurrence in patients who were diagnosed with suspicious recurrence by MRI after resection of supratentorial gliomas. Methods: A total of 164 patients (107 males, 57 females, age 6-74 years; high-grade 94, low-grade 63, unclear 7) with supratentorial gliomas who underwent ¹¹C-MET PET/CT between June 2015 and June 2017 in Beijing Tiantan Hospital were enrolled respectively. All patients were with suspicious recurrence after surgery showed by MRI and followed up for 6 months at least. The final diagnosis was determined with histopathological analysis or clinical follow-up. The maximum and mean standardized uptake value (SUV_max and SUV_mean), tumor-to-background ratios (TBR) of SUV_max and SUV_mean (TBR_max and TBR_mean) were recorded and compared between patients with recurrence or without recurrence using independent-sample t test. Receiver operating characteristic (ROC) curves were drawn to determine the threshold, and the diagnostic sensitivity and specificity of each parameter were calculated. Results: According to the clinical diagnosis, there were 139 patients with recurrence and 25 without recurrence. SUV_max, SUV_mean, TBR_max and TBR_mean were significantly higher for patients with recurrence than those without recurrence (4.19±1.95 vs 2.59±1.18, 2.34±1.08 vs 1.46±0.72, 2.95±1.17 vs 1.83±0.79, 2.64±1.11 vs 1.59±0.71; t values: 5.126-6.183, all P<0.01), but there was no difference in the areas under the ROC curve (AUC) for diagnosis of recurrence with the 4 parameters (z values: 0.265-1.674, all P>0.05), for which the optimal cut-off values were 3.05, 1.65, 1.96 and 1.79, respectively, and the corresponding sensitivities/specificities for the diagnosis of recurrence were 67.6%(94/139)/100%(25/25), 67.6%(94/139)/100%(25/25), 79.9%(111/139)/100%(25/25), 74.8%(104/139)/100%(25/25), respectively. Patients with (n=81) or without (n=13) recurrence had different semiquantitative parameters in high-grade glioma group (t values: 5.137-5.871, all P<0.01), and the AUC for TBR_mean was greater than that for SUV_mean (0.858 vs 0.802; z=1.982, P<0.05). Patients with (n=54) or without (n=9) recurrence in low-grade glioma group showed significant difference in the 4 parameters (t values: 2.730-7.009, all P<0.01), while the AUCs of the 4 parameters were not significantly different (z values: 0.444-1.407, all P>0.05). Among 37 patients with recurrence confirmed by pathology, there were no significant differences in the semiquantitative parameters between the high-grade and low-grade glioma groups and AUCs were not different either (t values: 1.387-1.937, z values: 0.106-1.752, all P>0.05). Conclusions Semiquantitative parameters of ¹¹C-MET PET/CT are equally accurate in the differentiation of recurrence from radiation injury in patients with gliomas, while TBR_mean was superior than SUV_mean in patients with the high-grade gliomas. Among the patients with recurrence confirmed by pathology, the value of the semiquantitative parameter is limited for the identification of high- and low- grade gliomas.

Objective To study the effect of early rehabilitation with full range of music therapy on acute coronary syndrome (ACS) patients after emergency PCI. Methods From July to December 2009, 108 inpatients undergone emergency PCI were randomly divided into experimental group and control group, with 54 patients in each group. The experimental group received full range of music therapy, which included PCI treatment under the circumstance of music and music therapy 3 times per day (10:00 a. m. , 16:00 p. m. and 21:00 p. m. ) during hospitalization. The control group received conventional treatment. SBP, HR, the left ventricular ejection fraction (LVEF) , 6-MWT, length of stay, the scale of anxiety and depression were compared between the two groups. Results The scores of anxiety and depression, SBP, HR, LVEF and 6-MWT in the experimental group were significantly different from those of the control group ( P < 0. 05). Conclusions Full range of music therapy significantly contributes to the rehabilitation of ACS patients after emergency PCI.

Objective To explore and compare the effectiveness,convenience and economic cost of two different methods of urokinase thrombolysis intermittently to recanalize PICC catheter.Methods 52 thrombotic completely blocked patients with PICC catheters were chosen and randomly divided into the experimental group and the control group,each with 26 cases.Both groups used the urokinase thrombolysis intermittently methods,the control group used "tee-vacuum suction" method while the experimental group used "injector-vacuum suction" method.Operation time for 10 suctions,results of thrombolysis,recanalization time,number of consumptive materials and economic cost were analyzed and compared between two groups.Results The recanalization rate was 96.15％ in the experimental group and 92.32％ in the control group,with statistically significant difference (x2 =0.517,P ＞ 0.05).The recanalization time and economic cost of consumptive materials were respectively (26.88 ± 18.30)min and (82.25 ± 0.61)yuan in the experimental group,(79.52 ±74.35)min and (90.86 ± 4.02)yuan in the control group,and the differences were statistically significant (t =8.22,3.44,10.59,respectively; P ＜ 0.01).Nurses' operative difficulty and feeling of hand fatigue were also lower in the experimental group than in the control group,and the difference was statistically significant (P ＜ 0.01).Conclusions "Injector-vacuum suction" is better than "tee-vacuum suction" as urokinase thrombolysis intermittently method.

This study was aimed to establish ELISA for recombinant bovine IFN-gamma (BovIFN-gamma) detection and provide a new method for diagnosis of pathogenic infection. The total RNA was isolated from peripheral blood leucocytes cultured with PHA mitogen stimulation. Then bovine IFN-gamma (BovIFN-gamma) gene cDNA was amplified by RT-PCR and cloned into pET28a to obtain the expression plasmid designated as pETBovIFN-gamma. The pETBovlFN-gamma was further transformed into competent E. coli BL21 cells and a 18kD His-tagged protein as expected was expressed after IPTG induction. By using purified recombinant BovIFN-gamma as antigen and lymphocyte-hybridoma technique, four hybridoma cell lines which stably secreted monoclonal antibodies against rBovIFN-gamma were generated, designated as A7, A10, G6, and G10. The immunoglobin subset was identified as IgG1 . Western-blotting analysis and ELISA demonstrated that the monoclonal antibodies secreted by all the four hybridoma cell lines could react specifically to the recombinant BovIFN-gamma, but not irrelative proteins such as Ag85B, ESAT-6-CFP-10 and GM-CSF, suggesting that the four hybridoma cell lines were rBovIFN-gamma specific monoclonal antibodies. A sandwich ELISA was established by using A10 secreted monoclonal antibody and rabbit polyclonal antibodies against BovIFN-gamma, HRP labeled goat anti-rabbit IgG. The results indicated that the sensitivity was 2ng/mL. This sandwich ELISA to detect BovIFN-gamma paved the way to develop a sensitive method for specific infection detection such as bovine tuberculosis diagnosis.
Animals ; Animals, Newborn ; Antibodies, Monoclonal ; biosynthesis ; immunology ; Antibody Specificity ; immunology ; Blotting, Western ; Cattle ; Cells, Cultured ; DNA, Complementary ; genetics ; Enzyme-Linked Immunosorbent Assay ; methods ; Female ; Hybridomas ; Interferon-gamma ; genetics ; immunology ; metabolism ; Mice ; Mice, Inbred BALB C ; Plasmids ; genetics ; Rabbits ; Recombinant Proteins ; immunology ; metabolism ; Reverse Transcriptase Polymerase Chain Reaction

Objective To study the detection methods of BK virus infection in kidney transplant recipients, and to explore the clinical application. Methods 132 cases of renal transplant recipients were undertaken BK virus detection including presence of decoy cells in urinary sediment, urine and serum BKVDNA to demonstrate the BK virus replication. Result Among 132 cases of renal transplant recipients,urinary decoy cell was found in 37 (28.0%)patients and the median time was 12 months after surgery. 32(24. 2% )patients were diagnosed as BK viruria at a median of 11 months after surgery, and 16( 12. 1% )recipients were diagnosed as BK viremia at a median of 15 months after surgery, 5 patients with BK viruria were diagnosed as BK virus associated nephropathy according to allograft biopsy. Conclusion To make early diagnosis of BK virus infection, detection of urine decoy cells and BKV-DNA in urine and plasma sample is important,which provides an important basis for the prevention of BK virus associated nephropathy.

Objective To explore the factors that affect the excretion rate of methotrexate and the occurrence of adverse reactions in high-dose methotrexate chemotherapy for osteosarcoma patients.Methods Retrospectively analyzed methotrexate excretion,liver injury,kidney injury,bone marrow suppression and other adverse drug reactions in 97 high-dose methotrexate chemotherapy cycles of 28 patients with osteosarcoma.The concentration of methotrexate in the blood at 0,24,48,72 h and the level of white protein in the blood were also analyzed.Results When the peak concentration of methotrexate(0h,Cmax)≥700 μmol·L-1 the risk of excretion delay increases:the incidence was 23.21％in group with Cmax ≥ 700 μmol·L-1,and it was 5.00％in group with Cmax＜700 μmol·L-1,(P＜0.05),but when the peak concentration was≥1 000 μmol·L-1,the risk of delayed excretion did not increase further:the incidence was 16.00％in group with Cmax 1 000 μmol·L-1,and it was 15.49％in group with Cmax＜1 000 μmol·L-1,(P＞0.05).Methotrexate blood Cmax has no significant correlation with the occurrence of important adverse reactions such as liver injury and bone marrow suppression.There was significant correlation between low serum albumin level and bone marrow suppression in patients.The average albumin level in group with bone marrow suppression was(39.1±3.4)g·L-1,which in without bone marrow suppression group was(41.2±4.0)g·L-1(P＜0.05).Conclusion During high-dose methotrexate chemotherapy in patients with osteosarcoma,delayed excretion and adverse reactions should not be prevented by lowering the peak concentration.The albumin level of patients is an important factor affecting the occurrence of bone marrow suppression.