OBJECTIVE:To improve the system of management and maintenance for the purification air-conditioning system in PIVAS,and to further strengthen the management of cleaning environment. METHODS:The cleanness monitoring project of purifi-cation air-conditioning system in PIVAS of our hospital was introduced in terms of temperature and humidity record,pressure differ-ence record,airborne particles detection,settling microbe monitoring report. And the monitoring results were analyzed. RESULTS:The temperature and humidity,pressure difference of clean area in PIVAS of our hospital are both in line with the standard of Phar-macy Intravenous Admixture Quality Management Specification (2010 edition),i.e. temperature at 18-26 ℃,relative humidity of 40%-65%;negative pressure difference between antibiotics,hazardous drug dispensing area and second dressing room are 5-10 Pa. The number of airborne particles (average static particle/m3) at various cleanness degrees in clean area are all in line with the standard of GMP(2010 edition),i.e. maximal allowable number of airborne particles(≥0.5 μm)were 3 520/m3(100 degree);352 000/m3 (10 000 degree);3 520 000/m3 (100 000 degree). The percentage of qualified static settling microbe detection reach 100%in clean area,which is in line with the standard of Settling Microbe Detection Method in Clean Room(Area) of Pharmaceu-tical Industry,i.e. criteria for settling microbe(90 mm)CFU/0.5 h≤1(100 degree);≤3(10 000 degree);≤10(100 000 degree). The percentage of qualified dynamic settling microbe detection is in low level,especially those of dispensing room and secondary dressing room only reaches 80%. CONCLUSIONS:It’s important for effective hospital infection control in PIVAS,the quality im-provement of intravenous injection,the safety guarantee of drug use in patients to further improve standard operation procedure of purification air-conditioning system management and maintenance,and manage and maintain the purification air-conditioning sys-tem completely and scientifically.

Objective To isolation and identify the pathogen of stem blight of Malvaceae.Methods The stems were collected from stem blight-diseased plants (J) and healthy ones (W) of Abelmoschus manihot (L.) Medic.in Yixing City of Jiangsu Province then cultured to isolate newborn mycelium.The pathogen isolated but unidentified were inoculated in stems of healthy plants of Abelmoschus manihot ( L.) Medic.and pathogenicity was verified.Finally, the pathogenic specie( s) was or were identified by morphological characteristic, rDNA-ITS analysis and polymerase chain reaction (PCR) method.Results The same fungus were excluded which were the same species in J and W, the three fungus of J2, J5 and J6 were acquired.J5 was preliminarily identified to have pathogenicity and it was Fusarium equiseti under the microscope.The genome DNA of J5 was amplified to a length of 524bp, and homology highly with Fusarium equiseti (100%).Conclusion The pathogen was identified as Fusarium equiseti.

Objective To investigate the demethylation and changes in gene expression of programmed death receptor-1 ( PD-1) caused by methylation inhibitor 5- azacytidine (5-Zac) in lymphocyte series Molt-4 cells and its mechanism. Methods Molt-4 cells were cultured in different concentrations of 5-Zac(0, 5, 10 Umol/L)for 72 h, ratio of cell expressing PD-1 and apoptosis rate were detected by FCM, transcription of PD-1 gene mRNA was detected by RT-PCR. Molt-4 cell DNA of all groups were disposed by sodium bisulfite, PD-1 gene promoter fragment binded with transcription factor Brn-2 was amplified by PCR,these amplification fragments were transformed into E. coli. Positive clones were selected by sequencing,methylation status of the fragments binded with transcription factor Brn-2 was examined. Results S-Zac could increase the PD-1 expression of Molt-4 cells. PD-1 expression rate in 0 μmol/L 5-Zac( 1. 13%±0.01% ) treated cells was found more lower than that in both 5 μmol/L and 10 μmol/L 5-Zac treated cells (18. 96% ±1. 87% , 63. 09% ± 6. 25% , P < 0. 05 ) , and they showed concentration-dependent (P <0.01). Cells apoptosis rate and PD-1 mRNA expression were also observed increased significantly with 5-Zac treating. Demethylation probability of CG points showed significant difference between transcription factor Brn-2 binding site and other four locations (P < 0.05 ). Conclusion 5 -Zac inhibits cell grouth in human lymphoid cell series Molt-4 by inducing PD-1 gene expression and promoter demethylation. PD-1 gene promoter binding transcription factor Brn-2 fragment CG point demethylation may be one of the important mechanisms in 5-Zac treated Molt-4 cells.

Objective To analyze the risk factors of respiratory depression occurring during recovery period in patients after having undergone general anesthesia and laparoscopic operation.Methods A total of 374 patients after general anesthesia and laparoscopic surgery admitted to the First Affiliated Hospital of Wenzhou Medical University from June 2015 to June 2016 were enrolled, they were divided into with or without the incidence of respiratory depression two groups by whether or not respiratorydepression, with the incidence of respiratory depression group 52 cases, without the incidence of respiratory depression group 322 cases. The patients' gender, age, body mass index (BMI), operation time, anesthesia maintenance mode, artificial airway mode, operative type and medication used in operation, intra-operative hypotension presence or absence, and type of operation were recorded. Univariate and multivariate logistic regression analyses were used to evaluate the risk factors of respiratory depression occurring in the recovery period after general anesthesia; receiver operating characteristic (ROC) curve was drawn to evaluate the predictive value of age, intraoperative medication, and age combine with intraoperative medication respectively in the occurrence of respiratory depression during recovery period after general anesthesia and lapatoscopic operation.Results Univariate analyses showed that there were no statistical significant differences in gender, BMI, operation time, anesthesia maintenance mode, artificial airway mode, intra-operative hypotension presence or absence, type of operation, etc. compared between patients with and without the incidence of respiratory depression groups (allP > 0.05); while the differences were statistically significant in age and drug used in the operation (dezocine, flurbiprofen, dexmedetomidine or dezocine combined with dexmedetomidine, allP < 0.05). Multivariate analyses showed that age and medication used in operation were the independent risk factors for the occurrence of respiratory depression during the anesthesia recovery stage (P values being 0.000, 0.002 respectively). ROC curve showed that age, intra-operative medication and age combine with intraoperative medication respectively had certain predictive value for the occurrence of respiratory depression during the recovery period after general anesthesia and laparoscopic surgery, the area under the ROC curve (AUC) of age combine with intraoperative medicationfor prediction of occurrence of respiratory depression during recovery period after anesthesia and laparoscopic surgery was significantly larger than that of single age or single intraoperative medication (0.826 vs. 0.668, 0.750,P < 0.01), 95% confidence interval (95%CI) of age, intraoperative medication and age combined with intraoperative medication were 0.598-0.738, 0.670-0.830, 0.764-0.888, the sensitivity, specificity and accuracy rate of age combine with intraoperative medication were 53.8%, 94.4% and 88.8%, respectively.Conclusion Elderly patients undergoing general anesthesia and laparoscopic operation and dezocine, dexmedetomidine or dezocine combined with dexmedetomidine being applied in the laparoscopic operation are more easily associated with incidence of respiratory depression during recovery period of anesthesia.

OBJECTIVE: To observe the demethylation effect of demethylation inhibitor 5-azacytidine (5-Zac) on programmed death receptor 1 (PD-1) in Molt-4 cells (T lymphocyte cell line) and to investigate the relationship between DNA demethylation and expression of PD-1. METHODS: Molt-4 cells were cultured in the medium containing different concentrations of 5-Zac(0, 5, 10 μmol/L) for 72 h. According to the concentrations of 5-Zac, the Molt-4 cells were divided into a 0 μmol/L 5-Zac group, a 5 μmol/L 5-Zac group, and a 10 μmol/L 5-Zac group. The expression of PD-1 in Molt-4 cells was detected by flow cytometry and the apoptosis rate was calculated. The mRNA transcription level of PD-1 was detected by real-time polymerase chain reaction; Molt-4 cell DNA in all groups were treated by sodium bisulfite. The PD-1 promoter fragment was amplified by PCR, the amplification fragments were transformed into E. coli., the positive clones were selected for equencing, and the methylation status of the fragments of PD-1 promoter was examined. RESULTS Seventy-two hours after the 5-Zac treatment, the expression rate of PD-1 in the Molt-4 cells in the 0 μmol/L 5-Zac group, the 5 μmol/L 5-Zac group, and the 10 μmol/L 5-Zac group was (1.13 ± 0.01)%, (18.96 ± 1.87)%, and (63.09 ± 6.25)% respectively, in a low concentration-dependent way. The PD-1 mRNA expression level was increased significantly with the 5-Zac treatment. Cells apoptosis showed that:compared with the 0 μmol/L 5-Zac group, the apoptosid rate in the 5 μmol/L 5-Zac group and 10 μmol/L 5-Zac group was signficantly increased, which was (1.9 ± 0.06)%, (8.89 ± 1.36)%, and (24.50 ± 3.68)% in the 0 μmol/L 5-Zac group, the 5 μmol/L 5-Zac group, and the 10 μmol/L 5-Zac mol/L group respectively. The bisulfite genomic sequencing showed that the demethylation probability of CpG points on -601 bp and -553 bp was significantly increased in the 5-Zac treated cells compared with those untreated. CONCLUSION 5-Zac can result in the increase of PD-1 expression in the human lymphoid cell series Molt-4 in vitro, and the apoptosis rate increases, which is related to PD-1 gene promoter demethylation.
Apoptosis ; genetics ; Azacitidine ; pharmacology ; Cell Line ; CpG Islands ; genetics ; DNA Methylation ; drug effects ; genetics ; Enzyme Inhibitors ; pharmacology ; Humans ; Programmed Cell Death 1 Receptor ; genetics ; metabolism ; Promoter Regions, Genetic ; genetics ; RNA, Messenger ; genetics ; metabolism ; T-Lymphocytes ; cytology ; metabolism

OBJECTIVE: To screen aptamers binding CD33+/CD34- cells from patients with acute myeloblastic leukemia M2 subtype (AML-M2). METHODS: CD33+/CD34- cells from patients with AML-M2 were taken as targeted cells, CD33+/ CD34- cells from normal people were taken as anti-selecting cells, and aptamers in the single strand deoxyribonucleic acid (ssDNA) library were then selected repeatedly by cell-systematic evolution of ligands by exponential enrichment (C-SELEX) technology, and amplified by polymerase chain reaction (PCR) to generate sub-ssDNA library. During the experiment, PCR amplification with fluorescently labeled primer and flow cytometry were performed to analyze the aptamers'enrichment of sub-library, and the final round product of the sub-ssDNA library was cloned. After the sequencing, the primary and secondary structures of the aptamers were analyzed. RESULTS: Electrophoresis indicated that the product of PCR amplification for each round subssDNA library was able to see a clear DNA band in the agarose gel. After 13 rounds of screening, the fluorescence intensity of the sub-ssDNA library binding the cells ranged from 2.14% to 51.12%, reaching a steady state at the 13th round. A total of 30 clones were selected and sequenced, 22 of which contained 1 of the 4 conserved sequences of AAGTA, TATCT, AGATG and AAATT in their primary structure, but the remained eight aptamers contained none of the conserved sequence. Secondary structure analysis indicated that four stem-loops and loop simulation convex structures existed in the aptamers. CONCLUSION C-SELEX technology can be used to screen the aptamers binding primary cells from patients with leukemia. The aptamers selected from the CD33+/CD34- cells from the patients of AML-M2 subtype might be used for the diagnosis and treatment for leukemia.
Adolescent ; Adult ; Antigens, CD34 ; genetics ; immunology ; Antigens, Differentiation, Myelomonocytic ; genetics ; immunology ; Aptamers, Nucleotide ; genetics ; metabolism ; DNA, Single-Stranded ; genetics ; Female ; Humans ; Leukemia, Myeloid, Acute ; genetics ; immunology ; Male ; Middle Aged ; SELEX Aptamer Technique ; Sialic Acid Binding Ig-like Lectin 3 ; genetics ; immunology ; Young Adult

ObjectiveTo investigate the clinical features and prognosis of culture-negative liver abscess (CNLA) versus culture-positive liver abscess (CPLA), and to provide a reference for early diagnosis and effective treatment. MethodsA retrospective analysis was performed for the clinical data of 371 patients with liver abscess who were admitted to The Second Affiliated Hospital of Air Force Medical University from 2005 to 2018, among whom 145 (39.1%) had positive results of pathogen test (CPLA group) and 226 (60.9%) had negative results (CNLA group). The two groups were compared in terms of clinical features, laboratory examination, imaging data, and prognosis. The t-test was used for comparison of normally distributed or approximately normally distributed continuous data between two groups, and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups; the chi-square test or the Fisher’s exact test was used for comparison of categorical data between two groups. ResultsCompared with the CNLA group, the CPLA group had a significantly older age (t=-3.464, P=0.001) and a significantly higher proportion of patients with diabetes (χ2=17.362, P＜0.001) or cardiovascular disease (χ2=10.827, P=0.001), and compared with the CPLA group, the CNLA group had a significantly higher proportion of patients with AIDS (χ2=4.354, P=0.037). Compared with the CNLA group, the CPLA group had significantly greater increases in leukocyte count, percentage of neutrophils, alkaline phosphatase, gamma-glutamyl transpeptidase, and total bilirubin (U=20393, 19711, 18586, 19349, and 18496, all P＜0.05), a significantly greater reduction in albumin (t=3.348, P=0.001), and a significantly higher proportion of patients with a baseline APACHE Ⅱ score of ≥16 (χ2=9.550, P=0.002). Compared with the CNLA group, the CPLA group had a significantly higher proportion of tumors with a diameter of ＞5 cm (χ2=53.61, P＜0.001). In the CNLA group, 19.9% of the patients were treated with anti-infective therapy alone, while for both groups, anti-infective therapy combined with ultrasound- or CT-guided percutaneous drainage was the main treatment method. There were no significant differences in the length of hospital stay, recurrence rate, and mortality rate between the two groups (all P＞0.05). ConclusionCompared with CPLA patients, CNLA patients tend to have a younger age and are less likely to develop severe inflammatory response and liver insufficiency, with a smaller diameter of abscess which is less likely to be treated by percutaneous drainage, while there is no significant difference in prognosis between CPLA patients and CNLA patients.

To study the molecular mechanism for DNA hypomethylation of STAT3 promoter in CD4+ T cells from acute graft-versus-host disease (aGVHD) patients.
 Methods: We collected CD4+ T cells from peripheral blood of 42 patients who underwent allogeneic hematopoietic stem cell transplantation (allo-HSCT) from HLA-identical sibling donors. GADD45A expression level in CD4+ T cells was measured by real-time PCR and Western blot. The binding level between HMGB1 and GADD45A in CD4+ T cells was analyzed by co-immunoprecipitation, while the binding levels of HMGB1/GADD45A with STAT3 promoter were detected by chromatin immunoprecipitation-quantitative real-time PCR (ChIP-qPCR). After overexpression of HMGB1 and knockdown of GADD45A in normal CD4+ T cells, STAT3 expression and DNA methylation were measured by Western blot and bisulfite sequencing PCR, respectively.
 Results: GADD45A expression was significantly up-regulated in patients with aGVHD compared with that in the patients without aGVHD. More HMGB1-GADD45A complexes were found in CD4+ T cells from patients with aGVHD compared with that in patients without aGVHD. The bindings of HMGB1/GADD45A with STAT3 promoter were significantly increased, and the binding levels of HMGB1/GADD45A were negatively correlated with STAT3 promoter DNA methylation. The expression of STAT3 was significantly reduced and the DNA methylation of STAT3 promoter was significantly increased in CD4+ T cells with overexpression of HMGB1 and knockdown of GADD45A compared with CD4+ T cells only with overexpression of HMGB1.
 Conclusion: The increased expression of HMGB1/GADD45A plays an importent role in STAT3 promoter DNA hypomethylation, thereby promoting STAT3 expression in CD4+ T cells from aGVHD patients.
CD4-Positive T-Lymphocytes ; Cell Cycle Proteins ; metabolism ; DNA Demethylation ; Gene Expression Regulation ; genetics ; Graft vs Host Disease ; genetics ; HMGB1 Protein ; metabolism ; Hematopoietic Stem Cell Transplantation ; Humans ; Nuclear Proteins ; metabolism ; Promoter Regions, Genetic ; genetics ; STAT3 Transcription Factor ; genetics ; metabolism

To demonstrate the effect of AB serum on terminal erythroid differentiation ex vivo.
 Methods: After separation of CD34+ cells from cord blood, the cells were cultured and divided into a control group and an experimental group. The effects of AB serum were examined by the expressions of different markers (GPA, Band3 and α4-integrin) for erythroblast differentiation and enucleation by flow cytometry. 
 Results: The CD34+ cells were successfully differentiated to enucleated red blood cells. There were evident differences among the expressions of GPA, Band3 and α4-integrin between the 2 groups. The percentage of GPA positive cells in the experimental group was bigger than that in the control group in every time point. The expression of Band3 in the experimental group was higher than that in the control group. The expression of α4-integrin in the experimental group was lower than that in the control group. In addition, the enucleation rate in the experimental group was higher than that in the control group.
 Conclusion: AB serum can promote the cell differentiation and enucleation during terminal erythroid differentiation in vitro.
ABO Blood-Group System ; blood ; physiology ; Anion Exchange Protein 1, Erythrocyte ; metabolism ; Antigens, CD34 ; blood ; Cell Differentiation ; genetics ; physiology ; Cell Nucleus ; Cells, Cultured ; Erythrocytes ; physiology ; ultrastructure ; Erythropoiesis ; genetics ; physiology ; Fetal Blood ; cytology ; physiology ; Flow Cytometry ; Glycophorins ; metabolism ; Humans ; Integrin alpha4beta1 ; metabolism

BACKGROUND: The 2019 novel coronavirus disease (COVID-19) emerges in China, which spreads rapidly and becomes a public health emergency of international concern. Chinese government has promptly taken quarantine measures to block the transmission of the COVID-19, which may cause deleterious consequences on everyone's behaviors and psychological health. Few studies have examined the associations between behavioral and mental health in different endemic areas. This study aimed to describe screen time (ST), physical activity (PA), and depressive symptoms, as well as their associations among Chinese college students according to different epidemic areas. METHODS: The study design is cross-sectional using online survey, from 4 to 12 February 2020, 14,789 college students accomplished this online study, participants who did not complete the questionnaire were excluded, and finally this study included 11,787 college students from China. RESULTS: The average age of participants was 20.51 ± 1.88 years. 57.1% of the college students were male. In total, 25.9% of college students reported depression symptoms. ST > 4 h/day was positively correlated with depressive symptoms (β = 0.48, 95%CI 0.37-0.59). COVID-19ST > 1 h/day was positively correlated with depressive symptoms (β = 0.54, 95%CI 0.43-0.65), compared with COVID-19ST ≤ 0.5 h/day. Compared with PA ≥ 3 day/week, PA < 3 day/week was positively associated with depression symptoms (β = 0.01, 95%CI 0.008-0.012). Compared with low ST and high PA, there was an interaction association between high ST and low PA on depression (β = 0.31, 95%CI 0.26-0.36). Compared with low COVID-19ST and high PA, there was an interaction association between high COVID-19ST and low PA on depression (β = 0.37, 95%CI 0.32-0.43). There were also current residence areas differences. CONCLUSIONS Our findings identified that high ST or low PA was positively associated with depressive symptoms independently, and there was also an interactive effect between ST and PA on depressive symptoms.
Adult ; COVID-19/psychology* ; China/epidemiology* ; Cross-Sectional Studies ; Depression/etiology* ; Exercise ; Female ; Humans ; Male ; Mental Health/statistics & numerical data* ; Screen Time ; Students/psychology* ; Time Factors ; Universities ; Young Adult