Objective To investigate the levels of serum chemerin in the acute myocardial infarction (AMI) patients and explore the clinical significance.Methods A total of 76 AMI patients from January 2013 to December 2013 were inrolled and 30 healthy patients were included as normal control(NC) group.Chemerin, tumor necrosis factor α(TNF-α), adiponectin were asessed by enzyme linked immunosorbent assay(ELlSA).The clinical data of high sensitivity C reactive protein (hs-CRP), body mass index (BMI), blood lipids, underlying diseases et al were collected.Results Serum levels of chemerin in AMI patients was higher than the NC group significantly((12.97±4.17) μg/L vs.(30.96± 17.16) μg/L, t =5.642, P＜0.001).The Serum levels of Chemerin in single vessel disease group, two vessel disease group and triple vessel disease group were (22.25 ± ±6.93) μg/L, (28.57 ± 15.92) μg/L, (37.95 ± 16.52) μg/L respectively and was linearly increasing relationship with increasing AMI severity (P＜0.001, F =22.84, r2 =0.397).Serum levels of Chemerin positively correlated with TNF-α, hs-CRP level(r2=0.347,0.455 ,P＜0.001) and negative correlated with adiponectin(r2 =0.396,P＜0.001).Logistic regression analysis showed that chemerin was an independent risk factor for AMI (OR =4.822,95％ CI 4.422-7.141, P =0.032).Conclusion The results showed that serum chemerin levels were significantly elevated in AMI patients,and chemerin may be involved in the development process of AMI.

Objective: To investigate the core information about tuberculosis prevention and control knowledge among residents in Jiangxi Province, so as to provide insights into formulation of health education strategy for tuberculosis prevention and control. Methods: Permanent residents at ages of 15 years and older were sampled from Jiangxi Province using a multi-stage stratified cluster sampling method from January to June 2021. Participants' demographic features and access to health education for tuberculosis prevention and control were collected through questionnaires, and the awareness of core information about tuberculosis prevention and control was investigated according to Key Points for Core Information and Knowledge about Tuberculosis Prevention and Control (2016 version). Results: A total of 1 280 questionnaires were allocated, and 1 217 valid questionnaires were recovered, with an effective recovery rate of 95.08%. The respondents included 605 men (49.71%) and 612 women (50.29%), and had a mean age of (50.05±15.78) years. The overall awareness of core information of tuberculosis prevention and control was 80.46%, and the awareness rates of “Pulmonary tuberculosis is suspected and timely healthcare-seeking is required if you have cough or expectoration for more than 2 weeks”, “Pulmonary tuberculosis is mainly transmitted via respiratory tract, and everyone is likely to be infected”, “No spit anywhere, covering mouth and nose when coughing or sneezing, and wearing a mouth mask may reduce the transmission of pulmonary tuberculosis”, “Pulmonary tuberculosis is a chronic infectious disease that poses a long-term damage to health” and “Most patients may be cured and others are avoided to be infected following standard whole-process treatment” were 90.22%, 86.52%, 85.95%, 80.03% and 59.57%, respectively. There were 491 respondents that were aware of all core information about tuberculosis prevention and control (40.35%), and network was the predominant route for acquiring health education about tuberculosis prevention and control (62.08%, 586/944). Conclusions The overall awareness of core information about tuberculosis prevention and control did not achieve the target set in the 13th Five-year Plan for Tuberculosis Prevention and Control, and the awareness of tuberculosis treatment-related knowledge was low.

Objective To explore the efficacy of multiplex ligation-dependent probe amplification (MLPA)combined with fluorescence in situ hybridization(FISH)and comparative genomie hybridization (CGH)combined with FISH in genetic analysis of chorionic villi specimen(CVS)of spontaneous abortion.Methods CGH+FISH and MLPA+FISH were used for genetic analysis of 29 CVS from spontaneous abortion and 6 normal CVS from selective abortion,in the mean time,those results were compared with conventional eytogenetic karyotyping.Results The report time were 40 hours in MLPA+FISH and 120 houm in CGH+FISH.The mean time of chorionic villi culture was(240±72)hours.The successful rate of specimen analysis were 97%(34/35)in CGH,100%(35/35)in MLPA,100%(35/35)in FISH and 91%(32/35)in conventional cytogenetic karyotyping.Apart from 1 case failed in CGH analysis,the results from MLPA+FISH were almost similar to that from CGH+FISH,however,that 1 specimen failed in CGH were detected successfully by MLPA+FISH.The discrepancy rate were 13%(4/31)in CGH+FISH and 12%(4/32)in MLPA+FISH respectively when compared with conventional cytogenetic analysis.Conclusions MLPA+FISH analysis present shorter detecting time and achieve 100%tale of successful report.This combined method was an important adjuvant approach to conventional cytogenetic karyotyping in CVS from spontaneous abortion.

Objective To study the immune system regulatory effects of CD8+CD28-regulatory T lymphocytes in an experimental bone marrow mesenchymal stem cell treatment of autoimmune encephalomyelitis.Methods A model of autoimmune encephalomyelitis (EAE) was established in twenty-eight C57BL/6 female mice,6 to 8 weeks old weighing 16 to 20 g using myelinoligodendrocyte glycoprotein 35-55 amino acid peptide (MOG35-55).The mice were randomly divided into a phosphate-buffered solution (PBS) group (n =7),an MSCs-Low group [n =7 which received an injection of 2× 105 mesenchymal stem cells (MSCs)],an MSCs-Med group (n=7,1× 106 MSCs),and an MSCs-High group (n=7,5×106 MSCs).Their clinical condition was then evaluated daily.On the 15th day after the MOG35-55 immunization,the different MSC doses were administered intravenously via the tail.On the 30th day the mice were sacrificed to observe any neuropathology of the spinal cord.At the same time,FACS flow cytometry was used to assay CD8+CD28-T cells in the spleen.Results Compared with the PBS control group,the MSC treatment effectively alleviated illness among the mice by the 15th day after the immunization.The maximum and average disease scores and clinical illness scores had all improved significantly.The medium dosage worked best.Neuropathological analysis showed that the MSC treatment could significantly reduce the invasion of inflammatory cells and minimize demyelination in the spinal cord.Furthermore,the CD8+ CD28-regulatory T cells in the spleens of the MSCtreated animals increased compared with the PBS control group,though the secreted levels of IL-10 showed no obvious difference.Conclusions Treatment with MCSs can promote the recovery of neural function in autoimmune encephalomyelitis,at least in mice.CD8+CD28-regulatory T cells may not be the main effector cells,playing only a synergistic therapeutic role.

Inthis study, the DNA fragment encoding the regions Ⅰ to Ⅱ of CSP gene from Plasmodium falciparm isolate FCC1/HN was cloned into an. expression vector pcDNA3 contained cytomegalovius (CMV) and transformed into human Hela cell line. The expressed protein PfCSP (condidate vaccine) was used to immunize BALB/c mice by subcutaneous、 intravenous or intraperitoneal administration respectively. The splenocyte of BALB/c mice immumized with the condidate vaccine released significantly IL-2 and IFN-γ following stimulation with this vaccine. It is associated with increase of the splenic T lymphocyte proliferation stimulated by this vaccine and enhancement of NK cell killing activity in the former studies. These results suggested that the vaccine could stimulate T cell response and enhance the cell-mediated immunity.

Objective To evaluate the value of ultrasound biomicroscopy (UBM) in assessment of atherosclerosis in apolipoprotein E (ApoE) knockout mice feeding with western diet.Methods Sixteen ApoE knockout mice in 8 weeks age were selected,then divided into two groups.One group was fed with west diet as high-fat group,and another group was fed with normal diet as control group.Intima-media thickness (IMT) and plaque area in the aortic root were assessed by UBM in two groups after 8 weeks and 16 weeks.And the measurements of UBM were compared with results of histopathology and blood-fat.ResultsThicken wall and plaque could be find in aortic root in control group and high-fat diet group byUBM.IMT and plaque area in high-fat diet group was significantly higher than those of control group ( P ＜ 0.05).The IMT and plaque area in UBM were good correlation with histopathology ( rwas 0.81 and 0.70 respectively).The triglyceride(TC) and total cholesterol in high-fat diet group was significantly higher than those of control group ( P ＜0.05),and IMT in UBM were increased with the elevated level of TC,there was a positive correlation between IMT and TC( r =0.528).ConclusionsWestern diet can accelerate the process in formation of atherosclerotic plaque in ApoE knockout mice.UBM can be used to observe this prograss noninvasively in vivo mice.

An analytical method was developed for the simultaneous extraction and determination of three quaternary ammonium compouds ( QACs) in soil samples using ultrasonic extraction and gas chromatography-mass spectrometry( GC-MS) . The qualitative and quantitative analysis of the three analytes dodecyltrimethyl ammonium chloride ( DTAC) , cetyltrimethyl ammonium bromide ( CTAB) and didodecyldimethyl ammonium chloride ( DDAC) were conducted by application of EI mass spectra and selected ion monioring ( SIM ) . Characteristic ions of the QACs were m/z 58 ( DTAC and CTAB) and m/z 212 ( DDAC) . To achieve optimum extraction efficiency, several impact factors including types of extractants, pH of extraction, concentration of linear alkylbenzene sulfonates ( LAS) , extraction times and content of purification column were investigated. Methanol with pH 3. 5 and 40 μg/L LAS solution were selected as extractant. Soil sample was extracted by treated methanol each 10 mL for 20 min every time. Extract of the soil sample was purified by neutral alumina column with 4 cm in length and 1cm in diameter, and then was determineted by GC-MS. Good linear relationships of all the three QACs were obtained in the range of 0. 02-2. 0 mg/L. The limits of determination (LOD, S/N=3) was 1. 2-4. 5 μg/kg. The method was used to analyse real soil samples ( paddy soil, lateritic red soil, and ore tailings) collected from a mining district in south China. Results of determination exhibited the concerntrations of the three analytes in real soil samples ranged from 0 . 24 mg/kg to 0. 41 mg/kg, and their recoveries ranged from 76% to 113% with relative standard deviations ( RSD) of 1. 1%-12. 9% in three different spiked concentrations of 0. 2, 0. 5 and 1. 0 mg/kg.

Objective:To investigate the effect of myeloid-derived suppressor cells (MDSC) on guanylate binding protein 1 (GBP1) in promoting the proliferation of glioma U87 cells and its mechanism.Methods:Glioma cells U87 with GBP1 overexpression (U87-GBP1) and control cells U87-Lacz transfected with empty vector were used as experimental cells. The mRNA and protein expressions of GBP1 and chemokine (C-C motif) ligand 2 (CCL2) in two groups of cells were detected by reverse transcription-quantitative polymerase chain reaction (RT-qPCR), Western blot and enzyme linked immunosorbent assay (ELISA), and the proliferation of U87 cells were detected by CCK-8. CD14 + monocytes and CD3 + T lymphocytes were isolated from peripheral blood of healthy people by immunomagnetic beads. The CD14 + monocytes were treated with culture medium of U87-Lacz cells or U87-GBP1 cells, and then the cells were divided into U87-Lacz culture medium group and U87-GBP1 culture medium group. The proportion of MDSC in CD14 + monocytes was analyzed by flow cytometry. CD14 + monocytes treated by two culture medium groups were cocultured with activated CD3 + T lymphocytes, and flow cytometry was used to detect the proliferation of activated CD3 + T lymphocytes. Monocytes untreated by U87 cells culture medium or activated CD3 + T lymphocytes were used as the control group. CD14 + monocytes were treated with U87-Lacz or U87-GBP1 cell culture medium anti-human CCL2 antibody, which were U87-Lacz+anti-CCL2 culture medium group and U87-GBP1 + anti-CCL2 culture medium group, and the proportion of MDSC in CD14 + monocytes was analyzed by flow cytometry. U87-GBP1 and U87-Lacz cells were inoculated into BALB/c nude mice to cause tumors in the brain. One week later, they were divided into chemokine (C-C motif) receptor 2 (CCR2) inhibitor RS504393 treatment group (U87-Lacz + RS nude mice group and U87-GBP1+RS nude mice group) and untreated control group (U87-Lacz nude mice group and U87-GBP1 nude mice group). After 30 days, the mice were sacrificed and the brain, spleen and bone marrow were isolated. Hematoxylin-eosin (HE) staining was used to observe the transplanted tumors in the brain of nude mice, and the volume of transplanted tumor was calculated, and flow cytometry was used to detect the proportion of MDSC in the tissues. Results:The protein expression of GBP1 in U87-GBP1 cells was significantly higher than that in U87-Lacz cells, but there was no significant difference in cell proliferation level between the two groups in vitro ( P > 0.05). The proportion of MDSC in U87-GBP1 culture medium group was significantly higher than that of U87-Lacz culture medium group [(7.75±0.80)% vs. (4.50±0.08)%], and both groups were higher than that of control group [(2.55±0.31)%)] ( F = 18.27, P = 0.002). The percentage of activated CD3 + T lymphocytes in U87-GBP1 culture medium group was lower than that in U87-Lacz culture medium group [(47.38±0.08)% vs. (61.70±5.05)%, P = 0.040]. The relative expression of CCL2 mRNA in U87-GBP1 cells and the expression level of CCL2 protein in U87-GBP1 cell culture medium [30.66±0.17 and (1 005.00±12.23) ng/L] were higher than those in U87-Lacz cells [1.29±0.15 and (111.60±11.44) ng/L] (both P < 0.01), the proportions of MDSC in U87-Lacz + anti-CCL2 culture medium group and U87-GBP1 + anti-CCL2 culture medium group was lower than those in U87-Lacz culture medium group and U87-GBP1 culture medium group (all P < 0.05). The volume of transplanted brain tumor in U87-GBP1 nude mice group was larger than that in U87-Lacz nude mice group; the volume of transplanted brain tumor in U87-GBP1 + RS nude mice group and U87-Lacz + RS nude mice group increased more slowly than the corresponding nude mice group without treatment, and the differences were statistically significant (all P < 0.05); the proportions of MDSC in transplanted brain tumor, spleen and bone marrow in U87-GBP1 nude mice group were higher than those in U87-Lacz nude mice group, and the proportions of MDSC in each tissue of U87-GBP1 + RS nude mice group and U87-Lacz + RS nude mice group were lower than those in the untreated by RS504393 corresponding nude mice group, and the differences were statistically significant (all P < 0.05). Conclusion:GBP1 might increase the expression of CCL2 in glioma U87 cells and recruit MDSC to form immunosuppression in glioma microenvironment, thus promoting the proliferation of glioma U87 cells in vivo.

Refractoriness of acute myeloid leukemia (AML) cells to chemotherapeutics represents a major clinical barrier. Suicide gene therapy for cancer has been attractive but with limited clinical efficacy. In this study, we investigated the potential application of herpes simplex virus thymidine kinase/ganciclovir (HSV-TK/GCV) based system to inhibit chemoresistant AML cells. We first generated Ara-C resistant K562 cells and doxorubicin-resistant THP-1 cells. We found that the HSV-TK/GCV anticancer system suppressed drug resistant leukemic cells in culture. Chemoresistant AML cell lines displayed similar sensitivity to HSV-TK/GCV. Moreover, HSV-TK/GCV killing of leukemic cells was augmented to a mild but significant extent by all-trans retinoic acid (ATRA) with concomitant upregulation of Connexin 43, a major component of gap junctions. Interestingly, HSV-TK/GCV killing was enhanced by expression of vesicular stomatitis virus G glycoprotein (VSV-G), a fusogenic membrane protein, which also increased leukemic cell fusion. Co-culture resistant cells expressing HSV-TK and cells stably transduced with VSV-G showed that expression of VSV-G could promote the bystander killing effect of HSV-TK/GCV. Furthermore, combination of HSV-TK/GCV with VSV-G plus ATRA produced more pronounced antileukemia effect. These results suggest that the HSV-TK/GCV system in combination with fusogenic membrane proteins and/or ATRA could provide a strategy to mitigate the chemoresistance of AML.
Cell Fusion ; Cell Line ; Coculture Techniques ; Connexin 43 ; Cytarabine ; Gap Junctions ; Genetic Therapy ; Glycoproteins* ; Homicide* ; K562 Cells ; Leukemia, Myeloid, Acute ; Membrane Proteins ; Simplexvirus* ; Suicide ; Thymidine* ; Tretinoin ; Up-Regulation ; Vesicular Stomatitis*

OBJECTIVETo explore the application of preimplantation genetic diagnosis (PGD) for infantile malignant osteopetrosis (IMO).

METHODSFor a family affected with IMO, PGD was provided using combined parental mutation detection and haplotype constructions with microsatellite markers spanning the TCIRG1 gene. Prenatal diagnosis was performed on the chorionic villus and amniocentesis samples by direct sequencing.

RESULTSPrenatal diagnosis showed that the fetus by the third pregnancy has carried the parental mutations [c.242delC (p.Pro81Argfs*85) and c.1114C>T (p.Gln372*)], and the pregnancy was terminated. PGD was subsequently performed through mutations detection and haplotype analyses following whole genome amplification (WGA) of each of 13 cells. The results showed that 6 of the 13 embryos were unaffected, 3 were carriers and 4 were affected. Well developed unaffected/carrier embryos were selected and transferred into the uterus. A single pregnancy was confirmed. Subsequently pre- and post-natal diagnoses have confirmed development of a healthy child.

CONCLUSIONThe study demonstrated the advantage of PGD over prenatal diagnosis when natural pregnancies have repeatedly produced IMO children/fetuses.

Adult ; Base Sequence ; Female ; Fertilization in Vitro ; Fetus ; Genetic Carrier Screening ; Humans ; Infant ; Male ; Microsatellite Repeats ; Molecular Sequence Data ; Osteopetrosis ; diagnosis ; embryology ; genetics ; prevention & control ; Pedigree ; Point Mutation ; Pregnancy ; Preimplantation Diagnosis ; Vacuolar Proton-Translocating ATPases ; genetics