Objective To evaluate preliminarily the significance of dynamic detection of Wilms'tumor gene(WT1)expression level on monitoring minimal residual disease(MRD)and predicting clinical relapse in patients of malignancy following allogeneic hematopoietic stem cell transplantation(allo-HSCT).Methods WT1 expression level WaS measured with real.time quantitative reverse transcription polymerase chain reaction(RQ-RT-PCR)method on 102 bone marrow specimens from 31 patients following allo.HSCT.WT1 expression level was determined as the ratio of WT1 mRNA to ABL mRNA times 100%.Resuits The levels of WT1 expression showed significant difference between the relapsed group and the non-relapsed group(P＜0.01),with 0.80%and 0.17%as their median expression level respectively.After dynamic measuring WT1 expression level on patients in the relapsed group.the level was found to increase significantly as compared with those during or before the clinical hematological relapse.both being＞1.0%.The level at the time of relapse Was significantly higher than the latest previous one(P＜0.05).Conclusion Dynamic detection of WT1 expression level with RQ-RT-PCR may be of help in monitoring MRD and warning clinical relapse Oil the patients following allo-HSCT.

Objective:To investigate the inhibitory effect of curcumin on oxidative stress in BCG-infected macrophages based on the Nrf2 pathway.Methods:THP-1-derived macrophages were infected.The experiment was divided into control group,BCG group,BCG+curcumin group and BCG+curcumin+ML385 group.Cellular ROS fluorescence intensity were observed under a fluores-cence microscope;Glutathione(GSH)levels were measured by Colorimetry;Western blot was used to detect the protein expressions of Nrf2,HO-1 and NQO1;MTT was used to detect the proliferation rate of macrophages.Results:BCG infection significantly enhanced ROS fluorescence intensity,reduced cell GSH content(P<0.01),inhibited protein expressions of Nrf2,HO-1 and NQO1,at the same time inhibited cell proliferation(P<0.01);curcumin significantly weakened ROS fluorescence intensity,increased GSH level(P<0.05),promoted Nrf2,HO-1 and NQO1 protein expressions and cell proliferation(P<0.01);Nrf2 inhibitor ML385 reversed the effect of curcumin.Conclusion:Curcumin can alleviate BCG-induced oxidative stress in macrophages by increasing the expression of Nrf2 and inducing the transcription of downstream antioxidant molecules.

OBJECTIVETo provide a quantitative summary in estimating the association between polymorphisms of 3 loci in NRAMP1 gene and susceptibility to tuberculosis in East-Asia population by means of meta-analysis.

METHODSWe searched databases (MEDLINE, OVID and CBM disc) from January 1995 to May 2005 using "NRAMP1" or "SLC11A1", in combination with "tuberculosis", also performed a manual search of citations from relevant original studies and literature. For each study involved, information was collected concerning the characteristics of the subjects, such as mean age of cases and the size of study. These characteristics were used to evaluate the sources of variation. Summary ORs and corresponding 95% CI were estimated by fixed effects (Mantel-Haenszel) or random effects (DerSimonian and Laird) model. To check for publication bias,a funnel plot, using Egger's linear regression method, was constructed. Cumulative meta-analysis was performed to evaluate whether the summary OR for studies with the polymorphisms of the 3 loci in the NRAMP1 gene was changing along with the accumulation of more data. Chi-square goodness of fit was used to test deviation from Hardy-Weinberg equilibrium.

RESULTSEight publications, with the number of cases and controls of 1067 and 1084 respectively, were identified and all genotype frequencies were consistent with Hardy-Weinberg equilibrium. The summary ORs for studies with polymorphisms of 3' UTR, D543N and INT4 loci of the NRAMPI gene among the East-Asia population were 1.68(95% CI: 1.31-2.16, P< 0.001), 1.78(95% CI: 1.38-2.30, P< 0.001), 1.56 (95% CI: 0.72- 3.35, P = 0.26), respectively when compared with their corresponding common homozygotes. Publication bias was not found in the studies with the three loci, except for INT4 locus, by Egger linear regression method. The cumulative summary effects ORs were 1.85 (P = 0.02) in 2000, 1.35 (P = 0.12) in 2002,1.64 (P= 0.001) in 2003, and 1.68 (P<0.001) in 2004 for 3'UTR locus, 1.88 (P = 0.001) in 2000,1.65(P = 0.001) in 2002,1.70(P<0.001) in 2003,1.76(P<0.001) in 2004, and 1.78(P<0.001) in 2005 for D543N locus, and 0.88(P = 0:70) in 2002, 2.50(P = 0.41) in 2003, 1.52(P = 0.42) in 2004 and 1.56(P = 0.26) in 2005 for INT4 loucs.

CONCLUSIONPolymorphisms at 3' UTR and D543N loci had statistically significant association between the NRAMP1 variants and susceptibility to tuberculosis in the East-Asia descendants, and variant in the INT4 locus failed to show statistically significant association in the East-Asia population.

Asian Continental Ancestry Group ; genetics ; Cation Transport Proteins ; genetics ; Far East ; Genetic Predisposition to Disease ; Humans ; Odds Ratio ; Polymorphism, Genetic ; Tuberculosis ; genetics

OBJECTIVETo establish a high-sensitivity, high-specificity and low-cost hydrogel chip platform for the clinical screening of Y chromosome microdeletions.

METHODSSite-specific extended primers with a common sequence at the 5' end were used for hybridizing with the target. The Cy5-dUTP was incorporated into the products by primer extension, and the products were labeled with fluorescence. Then the extended products were added to the chip for hybridizing with acrylamide-modified common probes immobilized on the chip. After removal of the free Cy5-dUTP by electrophoresis, the signals were obtained by fluorescence scanning. And the detecting conditions of this method were optimized.

RESULTSSY254 of 9 samples was successfully detected with the hydrogel chip. The results showed that 3 were normal and the other 6 with microdeletions (1 female sample as a negative control), which coincided with the results of conventional multiplex PCR-electrophoresis.

CONCLUSIONThe hydrogel chip platform we established has provided a new technique for the detection of Y chromosome microdeletions, and is beneficial to the diagnosis and treatment of male infertility.

Carbocyanines ; Chromosome Deletion ; Chromosomes, Human, Y ; genetics ; Deoxyuracil Nucleotides ; Humans ; Hydrogels ; Infertility, Male ; Male ; Oligonucleotide Array Sequence Analysis ; methods ; Sex Chromosome Aberrations ; Sex Chromosome Disorders of Sex Development ; diagnosis ; genetics

OBJECTIVETo observe the antimicrobial activity of topical agents commonly used for burns on Acinetobacter baumannii (AB) in both free and biofilm states, and their synergistic effect with ambroxol on AB within biofilm.

METHODSEleven AB strains were isolated from wound excretion, respiratory tract, and blood of patients hospitalized in our hospital from August 2005 to April 2007. (1) The minimal inhibitory concentration (MIC) and minimal bactericidal concentration (MBC) of mafenide acetate and chlorhexidine acetate to free AB (including drug-resistant, drug-sensitive, and standard strains) were determined by dilution method. (2) AB was cultured with LB or TSB medium for 12, 24, and 48 h to form biofilm, and it was treated with above-mentioned two topical agents in MBC (mafenide group and chlorhexidine group) for 30 min. Biofilm not treated by topical agent was used as control group. The biofilm thickness was determined with confocal laser scanning microscope. The proportion of living bacteria in biofilm was calculated. AB biofilm in each topical agent group was mixed and inoculated into LB culture dish to observe the growth of bacteria. (3) AB was cultured with LB medium for 48 h to form biofilm, which was respectively treated by above-mentioned two topical agents in MBC (mafenide group and chlorhexidine group) and combination of each topical agent with 3.75 mg/mL ambroxol solution (ambroxol + mafenide group and ambroxol + chlorhexidine group) for 30 min. Biofilm not treated by topical agents was used as control group. Growth of bacteria in biofilm was detected with MTT method (denoted as absorbance value). Data were processed with one-way analysis of variance and LSD-t test.

RESULTS(1) MIC of mafenide acetate and chlorhexidine acetate for free AB was respectively 25.00 mg/mL and 0.03 mg/mL. MBC of both agents for free AB was the same as their MIC. (2) Among three groups, the thickness of biofilm of sensitive AB was thicker than that of drug-resistant bacteria at most of the time points. Compared with those in control group, biofilm thickness and proportion of living bacteria in biofilm were slightly decreased in mafenide and chlorhexidine groups. The growth of bacteria was abundant in each group. (3) Absorbance value of drug-resistant bacteria in control, mafenide, and chlorhexidine groups was respectively 0.776 ± 0.071, 0.625 ± 0.063, and 0.420 ± 0.068. Absorbance value of drug-resistant bacteria in ambroxol + mafenide group (0.174 ± 0.089) was significantly lower than that of control group (t = 11.823, P = 0.000) and mafenide group (t = 9.248, P < 0.01). Absorbance value of ambroxol + chlorhexidine group (0.178 ± 0.044) was significantly lower than that of control group (t = 16.009, P = 0.000) and chlorhexidine group (t = 6.681, P < 0.01).

CONCLUSIONSDrug-resistant AB forms biofilm readily, which prevents topical agents from killing the bacteria inside. Combined use of ambroxol with topical agents gives synergistic effect on killing AB in biofilm in the wound.

Acinetobacter baumannii ; drug effects ; isolation & purification ; Anti-Bacterial Agents ; pharmacology ; Biofilms ; drug effects ; Burns ; microbiology ; Chlorhexidine ; pharmacology ; Drug Resistance, Bacterial ; Humans ; Mafenide ; pharmacology ; Microbial Sensitivity Tests

BACKGROUNDActive suppression by CD4+CD25+ regulatory T lymphocytes plays an important role in the down-regulation of T cell responses to foreign and self-antigens. This study was conducted to analyze whether the CD4+CD25+ regulatory T cells exist and function normally in tuberculous pleural effusion.

METHODSThe percentages of CD4+CD25+ T cells in pleural effusion and peripheral blood from patients with tuberculous pleurisy and peripheral blood from healthy control subjects were determined by flow cytometry. The expression of forkhead transcription factor Foxp3 was also examined. CD4+CD25+ and CD4+CD25(-) T cells from pleural effusion and blood were isolated, and were cultured to observe the effects of CD4+CD25+ T cells on proliferation response of CD4+CD25(-) T cells in vitro.

RESULTSThere were increased numbers of CD4+CD25+ T cells in tuberculous pleural effusion compared with peripheral blood from both patients with tuberculous pleurisy and normal subjects, and these cells demonstrated a constitutive high-level expression of Foxp3. Moreover, CD4+CD25+ T cells mediated potent inhibition of proliferation response of CD4+CD25(-) T cells.

CONCLUSIONThe increased CD4+CD25+ T cells in tuberculous pleural effusion express a high level of Foxp3 transcription factor, while potently suppressing the proliferation of CD4+CD25(-) T cells.

Adult ; Female ; Forkhead Transcription Factors ; analysis ; Humans ; Lymphocyte Activation ; Male ; Middle Aged ; Pleural Effusion ; etiology ; immunology ; T-Lymphocytes, Regulatory ; physiology ; Tuberculosis, Pleural ; etiology ; immunology

Objective To study the levels of serum cytokines in schizophrenic patients and their changes in antipsychotic medica-tion treatment .Methods The levels of serum cytokines including IL-10 ,IL-6 ,IL-13 ,IL-4 ,IFN ,TNF-α,IL-1a and IL-1RA were de-tected in 34 healthy adults and 53 schizophrenia patients by adopting the flow fluorescence method .Results The serum levels of IL-6 ,IL10 and TNF-αbefore treatment in schizophrenic patients were significantly higher than those in the control group (P<0 .05) . After treatment ,the levels of serum IL-1a ,IL-6 and TNF-α in schizophrenic patients were significantly lower than those before treatment(P<0 .05) .Conclusion Serum IL-6 and TNF-α levels are correlated with the disease condition of schizophrenia .IL-10 plays a role in early anti-inflammation of schizophrenia .

Objective To explore the impact of diagnosis related groups (DRGs) payment and drug zero plus on the management of intensive care medicine department. Methods The clinical data of patients in one year from 2016 to 2017 admitted into the Department of ICU in Liuzhou Worker's Hospital concerning their numbers of discharged patients, transferred patients, bed utilization rate, number of bed turnover, average length of stay of discharged patients, cure and improvement rates, admission and discharge diagnostic coincidence rate, 3-day definite diagnosis rate, clinicopathological diagnosis coincidence rate, rescue success rate, total income, drug proportion, consumable proportion, DRGs payment and settlement data, etc were retrospectively analyzed to explore the dual challenges, DRGs payment and drug zero plus, facing the department and how to respond and deal with them. Results In 2016 and 2017, the total incomes of the department of critical care medicine in our hospital were 42.107 0 million yuan and 41.371 3 million yuan respectively, and the medical insurance incomes were 15.03 million yuan and 16.69 million yuan respectively;in 2016 and 2017, 2 693 patients and 2 922 patients were admitted and treated respectively; 595 patients and 577 patients were discharged respectively, with 2 071 patients and 2 334 patients transferred respectively; the balances of the department were 15.48 million yuan and 29.11 million yuan, respectively. From July to December 2017, the medical insurance DRGs payment data suggested that the proportion of loss of the department be 7.02%. Accelerating the Grade 6 electronic medical records and informationization construction, adopting the severe disease information solution program and fine quality control management in the department of critical care medicine can reduce the cost of manpower. Conclusion Our future development direction in the Department of Intensive Care Medicine includes the following aspects: Open source and reduce expenditure, strictly control the proportions of drugs and consumables, improve the balance of the department, and actively respond and deal with the medical insurance DRGs payment.

OBJECTIVETo establish a model of acoustically evoked short latency negative response (ASNR) in guinea pigs, a model of profound hearing loss with normal saccular functions, and verify the correlation between ASNR and vestibular evoked myogenic potential (VEMP).

METHODSThirty-two healthy guinea pigs were employed in the experiment, which were randomly divided into control group (16 subjects) and deafened group (16 subjects). Each animal experienced auditory and vestibular tests including auditory brainstem response (ABR), VEMP and caloric test. A quick treatment was employed for deafened group consisting of a subcutaneous injection of kanamycin at a dose of 400 mg/kg followed by a jugular vein injection of ethacrynic acid at a dose of 40 mg/kg one hour later. The animals were received ABR, VEMP and caloric test 7 - 10 days following the drug administration. The deafened group was further divided into ASNR group and non-ASNR group, based on the presence of ASNR.

RESULTSIn deafened group, five subjects died postoperatively, 11 subjects (22 ears) provided full data, ASNR was elicited in eight ears (36.4%), the threshold was 120 - 130 dB SPL with mean of (124.4 ± 4.96) dB SPL. Its latency range was 1.75 - 2.60 ms with mean of (2.15 ± 0.27) ms. The mean latency of threshold was (2.34 ± 0.18) ms. All eight ASNR ears presented with VEMP. The VEMP threshold, positive and negative potential latencies proved no statistical difference (P > 0.05) between ASNR group and control group. Significant difference was detected between the VEMP presence of ASNR group and non-ASNR group (P = 0.002). There was no statistically significant correlation between VEMP and caloric test neither between ASNR and caloric test in deafened group.

CONCLUSIONSThis study evoked ASNR in an ototoxicity guinea pig model which has profound hearing loss with normal saccular functions. The presence of ASNR correlated with VEMP, however, not correlated with caloric test, suggesting that ASNR and VEMP are both originated from the saccule.

Animals ; Deafness ; physiopathology ; Disease Models, Animal ; Evoked Potentials, Auditory ; physiology ; Evoked Potentials, Auditory, Brain Stem ; Guinea Pigs ; Saccule and Utricle ; physiology ; Vestibular Evoked Myogenic Potentials ; Vestibular Function Tests

BACKGROUNDIn bone marrow transplant patients, the microenvironment in bone marrow is damaged after chemotherapy or radiotherapy. Subsequent to allogenic hematopoietic stem cell transplantation in patients with clinically successful engraftments, the source of mesenchymal stem cells (MSCs) remains controversial. To further verify the stimulatory effect of the simultaneous transplantation of cells from second donors on engraftment success for hematopoietic stem cell transplantation in support of donor MSCs engraftments, the aim of this study is to monitor the dynamics of the engraftment of bone marrow-derived MSCs in patients after transplantation with mismatched-sex hematopoietic stem and third-party cells.

METHODSIn this study, the hematopoietic stem cells from 32 clinical donors of different sexes that resulted in successful engraftments were selected for transplantation and were classified into three groups for research purposes: group A consisted of 14 cases of transplantation with bone marrow and recruited peripheral hematopoietic stem cell transplantation, group B contained 8 cases of simultaneous re-transfusion of MSCs from the second donor, and group C contained 10 cases of simultaneous re-transfusion of umbilical blood from the second donor. The bone marrow from 32 patients with successful engraftments of hematopoietic transplantation were selected and sub-cultured with MSCs. Flow cytometry (FCM) was used to measure the expression of surface antigens on MSCs. Denaturing high-performance liquid chromatography (DHPLC) in combination with polymerase chain reaction amplification of short tandem repeats (STRPCR) was used to measure the engraftment status of fifth-generation MSCs in patients. Fluorescence in situ hybridization (FISH) revealed the sex origin of the fifth-generation MSCs in 32 patients. Dynamic examinations were performed on patients receiving donor transplantations.

RESULTSThe progenies of fifth-generation MSCs were successfully cultured in 32 cases. The results of FCM demonstrated that the expression levels of CD14+ and CD45+ cells were lower than 0.04% in the fifth-generation MSCs. The analysis using DHPLC and FISH showed similar results. One patient from group B also received a temporary transplantation of MSCs from the donor. The MSCs in the remaining 31 patients all originated from the patients themselves.

CONCLUSIONSAfter transplantation, the MSCs present in patients originated from the host. In patients transplanted with MSCs from a second donor, the phenomenon of temporary chimerization of MSCs was observed.

Adolescent ; Adult ; Cells, Cultured ; Chromatography, High Pressure Liquid ; Female ; Flow Cytometry ; Hematopoietic Stem Cell Transplantation ; Humans ; Leukocyte Common Antigens ; metabolism ; Lipopolysaccharide Receptors ; metabolism ; Male ; Mesenchymal Stromal Cells ; cytology ; metabolism ; Middle Aged ; Young Adult