BACKGROUND:To improve the survival rate of transplanted hematopoietic stem cells, dynamic monitoring of plasma content of human cytomegalovirus (HCMV) and rapid screening of early active HCMV infection in hematopoietic stem celltranplantation recipients, thus to guide the clinical medication, is preferred. OBJECTIVE:To evaluate the usefulness of fluorescence quantitative PCR assay for early detection of HCMV activation after hematopoietic stem celltransplantation. METHODS:Real-time fluorescence quantitative PCR assay was applied for HCMV monitoring in 656 blood samples from 41 hematopoietic stem celltranplantation recipients and 60 control blood samples. RESULTS AND CONCLUSION:In 656 blood samples, 96 samples were positive, and the HCMV DNA copies ranged from 5.0×102 to 1.0×107 IU/mL. Timely initiation of therapy resulted in the rapid clearance of DNA-viraemia but it recurrenced in short time by drug-resistent. Two cases from 12 postive recipients died from HCMV infection. The quantitative detection of HCMV DNA by real-time PCR is a rapid method for monitoring HCMV infection and the early diagnosis in patients after hematopoietic stem celltransplantation.

Adolescent ; Child ; Female ; Humans ; Hypertension ; blood ; Insulin ; blood ; Insulin Resistance ; Male

Objective To explore the distribution of specific allergens for children in Sichuan Province and provide the evidences of clinical diagnosis and treatment.Methods Totally 451 cases in West China Second University Hospital,from April to August of 2016,were detected for 20 kinds of allergen-specific immunoglobulin E (IgE) by a new reverse enzyme-allergosorbent test,SPSS 18.0 statistical analysis was used to evaluate allergen-specific IgE levels in different groups of age,gender and diverse population distribution of disease.Results A total of 282 cases among 451 subjects were detected as IgE positive and the positive rate was 62.53%,among which 248 cases were positive for inhaled allergen-specific IgE,and the positive rate was 54.99%.There were 261 positive cases in food allergen-specific IgE,and the positive rate was 57.87%.For inhaled allergens,the highest positive rate was dermatophagoides pteronyssinus (n=111,24.61%),then dermatophagoides farina (n=110,24.39%),and house dust (n=108,23.95%);For food allergens,the highest positive rate was milk (n=159,35.25%),then egg (n=111,24.61%),and house dust (n=108,23.95%).Conclusions Both Inhaled allergens and food allergens were the important cause of allergic disease for children in Sichuan area.Serum allergen specific IgE determination was helpful to understand the patient′s allergy status,and diagnose the allergic disease.

Objective To study the effects of sample digestion conditions on measurement results of urinary iodine determined by As(Ⅲ)-Ce4+ catalytic spectrophotometry with ammonium persulfate digestion,and to promote the application of newly revised (the 2012 edition) national standard method for determination of urinary iodine.Methods According to the newly revised national standard method,various digestion conditions,such as ammonium persulfate concentration (0.8-1.3 mol/L,group interval 0.1),digestion instruments (heating block and drying oven) and standing time after digestion(0.5,1.0,2.0,4.0 and 22.0 h),were studied.The samples included 3 standard materials,which were GWB09108k,GWB09109f and GWB09110m containing iodine of (68.2 ± 9.0),(138.0 ± 10.0) and (221.0 ± 10.0) μg/L,and 5 urine samples with iodine concentration of 100-300 μg/L.Results Measurement results among the three groups of 0.9,1.0 and 1.1 mol/L ammonium persulfate digestion fluid showed no significant difference(P ＞ 0.05).The digestive effect showed no significant difference between heating block and drying oven (P ＞ 0.05) except one standard material in low concentration (GBW09108k).After digestion,samples were placed 0.5-22.0 h,the measurement results between groups showed no significant difference (P ＞ 0.05).Conclusions Appropriate concentrations of ammonium persulfate are from 0.9 mol/L to 1.1 mol/L.Heating block is recommended for the digestion,however,when absent,drying oven can be used alternatively.The standing times from 0.5 h to 22 h after digestion have not affected the measurement results.

Adolescent ; Adult ; Aged ; Endoscopy ; Female ; Humans ; Male ; Maxillary Sinus ; surgery ; Middle Aged ; Otorhinolaryngologic Surgical Procedures ; methods ; Young Adult

Objective To observe the effect of different intra-abdominal pressure and different time points on hemodynamics,ent-tidal CO_2(P_(ET)CO_2) and airway pressure(Paw) during the procedure of gynecological laparoscopic operations.Methods 60 cases undergoing gynecological laparoscopic operations were randomly divided into two groups:the intra-abdominal pressure was 1.3kPa in groupⅠ(30 cases) and 1.9kPa in groupⅡ(30 cases).ASAⅠgrade.In both groups,systolic blood pressure(SBP),diastolic blood pressure(DBP),mean arterial pressure(MAP), heart rates(HR).S-T.Paw and P_(ET)CO_2 were monitored and recorded before anesthesia(T_0),shortly after intubation (T_1),pre-pneumoperitoneum (T_2),5min after pneumoperitoneum (supine position) (T_3) and 5min (T_4),10min (T_5),20min(T_6),30min (T_7) after trendelenbury position (head down 200) and 5rain after deflation (T_8).Results In both groups SBP,DBP,MAP at time point T_3,T_4,T_5 were increased significantly compared with those of T_0 (P0.05),but there was significant difference in Paw and P_(ET)CO_2 in different time points within the same group and between the same time point in different groups after pneumoperitoneum(P

Aim To study targeting capability of anti-CD19 (Fab)-LDMto CD19 +B lymphoma cells in vi-vo and in vitro.Methods Flow cytometry was em-ployed to determine the affinity of Cy5 labeled anti-CD19 (Fab)-LDP to human lymphoma Raji cells.And the optical imaging system was used to analyze the dis-tribution of Cy5-anti-CD19 (Fab )-LDP in lymphoma-transplanted xenograft nude mice in vivo.Results The results of flow cytometry demonstrated that Cy5-an-ti-CD19(Fab)-LDP had remarkable affinity with lym-phoma Raji cells;Raji lymphoma xenograft model was established successfully in nude mice and in vivo fluo-rescence imaging analysis indicated that the antibody-drug conjugates could specially be localized in the tar-get tumor.Conclusion The experiments in vivo and vitro confirm that anti-CD19 (Fab)-LDP has remarka-ble affinity to targeting CD19 +lymphoma cells,and the antibody drugs anti-CD19 (Fab )-LDP have the probability to be new drugs for the treatment of malig-nant lymphoma.

The uracil in DNA comes from either the misincorporation of dUTP in place of dTTP or deamination of cytosine. In the latter case, it can result in a GC to AT transition mutation if the uracil is not removed before DNA replication. Base excision repair (BER) is a major pathway for removing DNA lesions arising from endogenous processes as well as those induced by exposure to exogenous chemicals or irradiation. BER is initiated by DNA glycosylases that excise aberrant bases from DNA by cleavage of the N-glycosidic bond linking to the base of its deoxyribose sugar. Uracil N-glycosylase (UNG) is the enzyme responsible for the first step in the BER pathway that specifically removes uracil from DNA. The UNG gene undergoes both temporal and spatial regulation mainly at the level of transcription. Normally cancer cells undergo over-proliferation and up-regulate their UNG during tumorigenesis. In this study we examine the correlation between UNG level and carcinogenesis, and explore the possibility of using UNG as a marker for cancer diagnosis. Human UNG gene was amplified from the total RNA of the human choriocarcinoma cell line, JEG-3, by RT-PCR. After purification, the 942bp full-length UNG cDNA coding sequence was digested with EcoR I and Sal I, and cloned into the digested pET-21 to construct a recombinant vector, pUNG. The UNG protein was expressed under the control of T7 promoter in E. coli BL21 (DE3) cells induced with IPTG. After ultrasonic treatment, the cell lysate and precipitate were analyzed by SDS-PAGE and a 39kD band was detected. The plasmid was serially diluted at appropriate concentrations and employed as standards in the subsequent quantification. Total RNAs were extracted from 18 pairs of clinical samples, each pair contains a sample of esophageal squamous cell carcinoma (ESCC) tissue and its surrounding normal esophageal epithelia. The copy numbers of UNG mRNA in these RNA samples were determined by real-time quantitative RT-PCR using a Lightcycler (Roche). UNG was present in 13 cases of ESCC (13/18, n = 18) but absent in all of the normal tissues. The results indicated that there was a correlation between high level of UNG expression and the carcinogenesis of ESCC.
Carcinoma, Squamous Cell ; genetics ; metabolism ; Cell Line, Tumor ; Cloning, Molecular ; Electrophoresis, Polyacrylamide Gel ; Esophageal Neoplasms ; genetics ; metabolism ; Humans ; In Vitro Techniques ; Reverse Transcriptase Polymerase Chain Reaction ; Uracil-DNA Glycosidase ; genetics ; metabolism

Based on the different permeability of DNA-intercalant dyes YO-PRO-1(YP) and propidium iodide (PI) to the membrane of viable, apoptotic and necrotic cells, cell samples were stained with 4?mol/L YP and 4?g/ml PI for 10 min, and the fluorescence intensity of both YP and PI were measured by fluorometer at Ex/Em wavelength of 485/538nm and 530/590nm, respectively. The correlation between YP fluorescence intensity and the apoptotic cell number was confirmed by fluorescence microscope and linear regression(r=0.999,P

OBJECTIVETo investigate the expression and procoagulant activity of phosphatidylserine (PS) on the normal peripheral blood cells of adults.

METHODSNormal peripheral blood samples were collected from 10 healthy volunteers (5 ml from each volunteer), platelets, neutrophils, lymphocytes and erythrocytes were isolated. The expression and procoagulant activity of PS on normal blood cells were identified by flow cytometry, inhibition test with lactadherin as PS probe and coagulation anticoagulant, respectively.

RESULTSThere was PS expression on a few normal blood cells (9.1%, 5.4%, 3.9% and 3.2% in platelets, neutrophils, lymphocytes and erythrocytes, respectively). The PS on these normal blood cells in vitro showed significant procoagulant activity. The plasma recalcification time was shortened by 47%, 36.5%, 25% and 12.5% by platelets, neutrophils, lymphocytes and erythrocytes, respectively; the formation of factor Xa (through both intrinsic and extrinsic pathways) and thrombin was also increased by 13% - 26% by platelets, neutrophils, lymphocytes and erythrocytes, respectively.

CONCLUSIONThe PS on normal blood cells in vivo may play a crucial role in the coagulation cascade.

Adult ; Blood Cells ; metabolism ; physiology ; Blood Coagulation Tests ; Female ; Flow Cytometry ; Humans ; Male ; Phosphatidylserines ; metabolism