1.Distribution and influence factors of Anammox bacteria in sewage treatment systems.
Bingyu ZHENG ; Yongzhen PENG ; Liang ZHANG ; Anming YANG ; Shujun ZHANG
Chinese Journal of Biotechnology 2014;30(12):1817-1827
Nitrogen removal techniques based on Anammox process are developing rapidly these years. The distribution and diversity of Anammox have become important research directions. A variety of Anammox have been detected till now, of which only Kuenenia and Brocadia are often detected in wastewater treatment systems. In addition, in a single niche there is only one type of Anammox bacteria. However, the distribution mechanism and transformation of Anammox bacteria in different niches are still ambiguous. Therefore, the distribution of Anammox in various conditions was summarized and analyzed in this article. And the key factors influencing the distribution of Anammox were concluded, including substrate concentration and the specific growth rate, sludge properties and microbial niche, the joint action and influence of multiple factors. The engineering significance research on the distribution and influencing factors of Anammox bacteria in the sewage system and proposed research prospects were expounded.
Ammonia
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chemistry
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Anaerobiosis
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Bacteria
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Nitrogen
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chemistry
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RNA, Ribosomal, 16S
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Sewage
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microbiology
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Waste Disposal, Fluid
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Waste Water
2.Effects of autologous blood withdrawal-reinfusion on perioperative coagulation function in patients undergoing cardiac surgery with cardiopulmonary bypass
Sheliang SHEN ; Yihong XIE ; Bingyu CHEN ; Yongjian CHEN ; Jinju GUAN ; Jiayin ZHENG
Chinese Journal of Anesthesiology 2014;(3):270-274
Objective To investigate the effects of autologous blood withdrawal-reinfusion on the perioperative coagulation function in patients undergoing cardiac surgery with cardiopulmonary bypass (CPB ) . Methods Eighty-four ASA physical status Ⅱ-Ⅳ patients ,without impairment of coagulation function ,scheduled for cardiac surgery with CPB ,were equally and randomly divided into 2 groups using a random number table :autologous blood withdrawal-reinfusion group (group ABWR , n= 44 ) and control group (group C , n= 40 ) . Decreased coagulation function was diagnosed based on the following two criteria :laboratory standard of decreased coagulation function and clinical signs .After anesthesia and before the beginning of operation (T1 ) ,at 5 min after heparin was reversed with protamine (T2 ) ,at the end of operation (T3 ) and at 24 h after the end of operation (T4 ) ,venous blood samples were obtained to measure the blood routine and parameters of coagulation function . Blood routine included the red blood cell (RBC ) , hemoglobin (Hb ) , hematocrit (Hct ) , platelet count , and plasma fibrinogen concentration (Fib) .The parameters of coagulation function included thrombelastography (TEG) variables and prothrombin time (PT ) ,activated partial thromboplastin time (APTT ) ,international normalized ratio (INR ) , and activated clotting time (ACT ) . The volume of intraoperative blood loss , amount of mediastinal drainage at 6 and 24 h after operation , consumption of tranexamic acid and heparin during operation , and consumption of fibrinogen after operation ,and requirement for transfusion of allogeneic RBCs ,fresh frozen plasma (FFP) and platelet during operation and within 24 h after operation were recorded .The development of decreased coagulation function during operation and within 24 h after operation .Results Compared with group C , perioperative consumption of allogeneic RBCs were decreased ,reaction time (R) measured by celite-activated TEG was increased at T3 (P<0.05) ,and no significant change was found in the blood routine index ,incidence of decreased coagulation function ,volume of intraoperative blood loss ,amount of mediastinal drainage ,consumption of tranexamic acid and heparin , and postoperative consumption of fibrinogen in group ABWR ( P> 0.05 ) . Conclusion Autologous blood withdrawal-reinfusion provides similar effects on coagulation function with allogeneic blood transfusion ,and does not increase the development of decreased coagulation function in patients undergoing cardiac surgery with CPB .
3.Effect of homogeneous management combined with staged teaching on physicians receiving standardized training of hepatobiliary surgery
Bingyu XING ; Jie GAO ; Yaguang HU ; Cunyi SHEN ; Qinling YANG ; Zheng WANG ; Wenjun TAN
Chinese Journal of Medical Education Research 2023;22(7):1059-1062
Objective:To investigate the effect of homogeneous management combined with staged teaching on physicians receiving standardized training of hepatobiliary surgery.Methods:A total of 46 physicians who received standardized training in Department of Hepatobiliary Surgery, The First Affiliated Hospital of Xi'an Jiaotong University, from January to March 2020 were selected as control group and were given conventional teaching, and 50 physicians who received standardized training from April to June 2020 were selected as observation group and were given homogeneous management combined with staged teaching. The two groups were compared in terms of professional level, clinical ability, and the degree of satisfaction with teaching before and after teaching. SPSS 24.0 was used to perform the independent samples t-test, the paired t-test, the chi-square test, and the rank sum test. Results:After teaching, both groups had significant increases in the scores of theoretical examination and operation skill examination, and compared with the control group, the observation group had significantly higher scores of theoretical examination (94.57±3.28 vs. 90.32±2.12) and operation skill examination (94.37±4.18 vs. 91.25±3.46). After teaching, both groups had significant increases in the scores of clinical consultation, physical examination, humanistic concern, clinical diagnosis, communication ability, organizational ability, and overall evaluation, and the observation group had significantly higher scores of the above seven aspects than the control group (6.98±0.94/6.45±0.14/6.95±0.88/6.65±0.93/6.53±0.26/6.84±0.92/6.58±0.35 vs. 6.13±0.31/6.21±0.76/6.21±0.42/6.18±0.35/6.32±0.61/6.33±0.24/6.25±0.71). The observation group had a significantly higher overall satisfaction rate than the control group [94.00% (47/50) vs. 78.26% (36/46)].Conclusion:In the standardized training and teaching of hepatobiliary surgery, homogeneous management combined with staged teaching can improve the professional level and clinical ability of physicians and enhance the degree of satisfaction with teaching.
4.Analysis of a child with X-linked intellectual disability due to a maternal de novo splicing variant of the PAK3 gene.
Chen WANG ; Xueping QIU ; Hui HU ; Bingyu JIN ; Yating CHENG ; Yue ZHAO ; Chun ZHOU ; Ling MA ; Yuanzhen ZHANG ; Fang ZHENG
Chinese Journal of Medical Genetics 2023;40(7):865-870
OBJECTIVE:
To explore the genetic etiology for a child with profound intellectual disabilities and obvious behavioral abnormalities.
METHODS:
A male child who had presented at the Zhongnan Hospital of Wuhan University on December 2, 2020 was selected as the study subject. Peripheral blood samples of the child and his parents were collected and subjected to whole exome sequencing (WES). Candidate variant was verified by Sanger sequencing. Short tandem repeat (STR) analysis was carried out to determine its parental origin. The splicing variant was also validated in vitro with a minigene assay.
RESULTS:
WES results revealed that the child had harbored a novel splicing variant of c.176-2A>G in the PAK3 gene, which was inherited from his mother. The results of minigene assay have confirmed aberrant splicing of exon 2. According to the guidelines from the American College of Medical Genetics and Genomics, it was classified as a pathogenic variant (PVS1+PM2_Supporting+PP3).
CONCLUSION
The novel splicing variant c.176-2A>G of the PAK3 gene probably underlay the disorder in this child. Above finding has expanded the variation spectrum of the PAK3 gene and provided a basis for genetic counseling and prenatal diagnosis for this family.
Child
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Female
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Humans
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Male
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Pregnancy
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Exons
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Intellectual Disability/genetics*
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Mothers
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Mutation
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p21-Activated Kinases/genetics*
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Parents
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RNA Splicing
5.Research progress of ICU-acquired weakness
Hui ZHENG ; Yuan SHI ; Zhaolong ZHANG ; Danyang ZHAO ; Congyi ZHAO ; Bingyu QIN
Chinese Critical Care Medicine 2024;36(3):308-312
ICU-acquired weakness (ICU-AW) is a common complication in the intensive care unit (ICU). The occurrence of ICU-AW directly leads to prolonged ICU stays for critically ill patients, and in severe cases, it continues to affect their quality of life even after discharge. This article provides a comprehensive review of the research progress on ICU-AW based on domestic and foreign studies, aiming to provide a scientific overview of ICU-AW, including its definition, pathophysiology, diagnosis, screening tools, influencing factors, and potential intervention strategies, so as to promote timely planning and implementation of relevant screening and intervention measures.
6.The construction of anti-CD19 chimeric receptor modified NK-92 cells and the killing effect of CD19 positive non-Hodgkin lymphoma cells
JIANG Xin ; ZHU Shuangyue ; ZHENG Haili ; LIU Bingyu
Chinese Journal of Cancer Biotherapy 2018;25(8):767-771
Objective: A second generation CAR-NK-92 cell line expressing CD19 was constructed to investigate its specific killing effect on CD19 positive non-Hodgkin lymphoma cells. Methods: First, build CD19-CAR gene expression vector and packaged slow virus particles, then the infection rate was detected by flow cytometry after infected NK-92 cells and positive cells were further separated. Finally, detected the expression of CD19-CAR in NK-92 cells by Western blotting. U-266 with CD19 negative myeloma cells,ARH77 and HS-Sultan with CD19 positive non-Hodgkin’s lymphoma cells as target cells, and CD19CAR-NK-92 as effector cells, then the killing rate was calculated by the absolute number of tumor cells alive in the cell killing experiment. Results: Construct lentivirus vector pLVX-CD19-CAR and packaged virus particles successfully, the purity of CD19-CAR-NK-92 cells also was over 90% after infected with NK-92 cells; and Western blotting analysis showed that CD19-CAR had been successfully expressed in NK-92 cell. The killing effect of CD19CAR-NK-92 onARH-77 ([70.10±1.86]% vs [1.95±0.63]%, P<0.01) and HS-Sultan ([74.98±1.60]% vs [0.58±1.49]%, P< 0.01) cells was significantly higher than the empty vector control group of ZsGreen-NK-92, but there was no difference in killing U266 (P>0.05). Conclusion: The NK-92 cell lines expressing CD19CAR were successfully constructed, and also has specific killing effects on CD19 positive non-Hodgkin lymphoma cells.
7.Xuebijing injection improve pulmonary vascular barrier function in ARDS by up-regulating claudin-5 expression through PI3K/Akt/FOXO1 signaling pathway
Ping GENG ; Jiali XIONG ; Fen YU ; Huihui WANG ; Ying WANG ; Min XU ; Bingyu LING ; Aiwen MA ; Ruiqiang ZHENG
Chinese Critical Care Medicine 2022;34(2):145-150
Objective:To study the signaling pathway of the up-regulation of claudin-5 expression by Xuebijing injection.Methods:Animal and cell models of acute respiratory distress syndrome (ARDS) were induced by lipopolysaccharide (LPS). ① In vivo study, 20 male Sprague-Dawley (SD) rats were randomly divided into 4 groups: control group, LPS group (LPS injection 10 mg/kg for 12 hours), Xuebijing control group (Xuebijing injection 1 mg/kg, twice a day, for 3 days), and Xuebijing intervention group (LPS injection after pretreatment of Xuebijing injection), according to random number method with 5 rats in each group. The lung tissues were taken to detect lung dry/wet weight ratio (W/D) and the morphological changes in each group. Claudin-5, phosphorylated forkhead box transcription factor O1 (p-FOXO1), total FOXO1 (t-FOXO1), phosphorylated Akt (p-Akt) and total Akt (t-Akt) in lung tissues were detected by immunohistochemical staining (IHC) and Western blotting. ② In vitro study, human pulmonary microvascular endothelial cells (HPMECs) were divided into 6 groups (5 holes in each group): control group, Xubijing control group (incubated with 2 g/L Xubijing for 24 hours), phosphoinositide 3-kinases (PI3K) signaling pathway LY294002 control group (incubated with 10 μmol/L LY294002 for 1 hour), LPS group (incubated with 1 mg/L LPS for 12 hours), Xubijing intervention group (incubated with 2 g/L Xuebijing for 24 hours, then with 1 mg/L LPS for 12 hours) and LY294002 intervention group (incubated with 10 μmol/L LY294002 for 1 hour, then with 2 g/L and Xubijing for 24 hours, and then with 1 mg/L LPS for 12 hours). The expression levels of claudin-5, p-FOXO1, t-FOXO1, p-Akt and t-Akt of HPMECs in each group were assessed by Western blotting. Results:In vivo study: ① Compared with the control group, the lung W/D ratio increased significantly in LPS group (6.79±0.42 vs. 4.19±0.13), and decreased significantly after the intervention of Xuebijing (4.92±0.38 vs. 6.79±0.42, P < 0.01). ② Morphological changes of lung tissue: compared with the control group, the injury of lung tissue in LPS group was more serious, which was significantly improved after Xuebijing intervention. ③ Expression levels of claudin-5, p-Akt/t-Akt and p-FOXO1/t-FOXO1: the expression levels of claudin-5, p-Akt/t-Akt and p-FOXO1/t-FOXO1 in LPS group were significantly decreased as compared with the control group (claudin-5/GAPDH: 0.33±0.03 vs. 1.03±0.07, p-Akt/t-Akt: 0.18±0.02 vs. 1.01±0.13, p-FOXO1/t-FOXO1: 0.16±0.06 vs. 1.00±0.19, all P < 0.01). After the intervention of Xuebijing, the expression levels were significantly increased as compared with the LPS group (claudin-5/GAPDH: 0.53±0.05 vs. 0.33±0.03, p-Akt/t-Akt: 0.56±0.12 vs. 0.18±0.02, p-FOXO1/t-FOXO1: 0.68±0.10 vs. 0.16±0.06, all P < 0.01). In vitro study: compared with the control group, the expression level of claudin-5 in the LPS group was significantly decreased (claudin-5/β-actin: 0.45±0.03 vs. 1.01±0.15, P < 0.01), and the expression level of claudin-5 in Xuebijing intervention group was also significantly decreased (claudin-5/β-actin: 0.80±0.08 vs. 1.01±0.15, P < 0.01). After the intervention of LY294002, the expression of claudin-5 was significantly decreased as compared with the Xubijing intervention group (claudin-5/β-actin: 0.41±0.02 vs. 0.80±0.08, P < 0.01). Conclusion:Xuebijing injection improve pulmonary vascular barrier function in rats with ARDS by up-regulating claudin-5 expression through PI3K/Akt/FOXO1 signaling pathway.
8. The effect of PM2.5 on oncogene expression in HBE cells
Kai ZHENG ; Bingyu WANG ; Xinyun XU ; Hong GENG ; Dingxin LONG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2019;37(8):561-566
Objective:
To study the effect of particulate matter 2.5 (PM2.5) on oncogene expression in human bronchial epithelial (HBE) cells.
Methods:
HBE cells were selected as the study subjects, and PM2.5 treatment group (10 μg/ml and 50 μg/ml) , negative control group and positive control group (10 μmol/L Cr6+) were set. CCK8 assay was used to test the IC50 value of PM2.5. HBE cells were treated with PM2.5 for 24 h at 10 μg/ml and 50 μg/ml, additionally, cells were treated with blank as negative control, 10 μmol/L Cr6+ as a positive control for 24 h. After the treatment, mRNA expression of oncogenes including c-myc, c-fos, k-ras and p53 were detected by fluorescent quantitative RT-PCR, the protein expression of oncogenes were detected with western blot.
Results:
The IC50 value of PM2.5 in HBE cells is 70.12 μg/ml. The qRT-PCR data showed that compared with the control group, the expression level of c-myc gene increased by respectively 500.1%、780.7%、305.3% after exposure to 10、50 μg/ml PM2.5 and positive control group; c-fos gene increased respectively 34.0%、76.7%、131.3% after exposure to 10、50 μg/ml PM2.5 and positive control group; k-ras gene increased respectively 50.3%、107.0%、49.7% after exposure to 10、50 μg/ml PM2.5 and positive control group; p53 gene decreased by 28.3%、28.7%、59.7% after exposure to 10、50 μg/ml PM2.5 and positive control group. The western blot results showed that compared with the control group, c-myc protein increased respectively 29.7%、77.3% after exposure to 50 μg/ml PM2.5 and positive control group; c-fos protein increased respectively 200.3%、137.0% after exposure to 50 μg/ml PM2.5 and positive control group; k-ras protein increased respectively 106.3%、130.3%、116.7% after exposure to 10、50 μg/ml PM2.5 and positive control group; p53 protein decreased by 43.7%、53.3%、52.1% after exposure to 10、50 μg/ml PM2.5 and positive control group.
Conclusion
PM2.5 could promote the expression of oncogenes in HBE cells, the carcinogenicity of haze might be related to promotion of oncogenes expression induced by PM2.5.
9. Construction of 3β-HSD gene silencing cell line and its effects on apoptosis induced by DEHP
Li WANG ; Xiaoyun QIN ; Xinyun XU ; Huaicai ZENG ; Kai ZHENG ; Bingyu WANG ; Dingxin LONG
Chinese Journal of Industrial Hygiene and Occupational Diseases 2019;37(1):13-19
Objective:
To construct 3β-HSD gene shRNA lentivirus interference vecto, then transfect into human MCF-7 cells, and construct cell line with 3β-HSD gene silencing, finally to study the effects of 3β-HSD on apoptosis induced by di- (2-ethylhexyl) phthalate (DEHP) .
Methods:
According to the mRNA sequence of 3β-HSD gene provided by GenBank, three interference sequences were designed and connected to PLVX-shRNA2-puro after annealing. The recombinant lentivirus vector was transfected into 293FT cells, the virus supernatants were collected and infected with MCF-7 cells. After puromycin screening, MCF-7 cells with 3β-HSD gene silencing were constructed. The cells with 3β-HSD gene silencing were identified by real-time quantitative PCR and western blot. Then the 3β-HSD gene silencing cells and MCF-7 cells were treated at various doses of DEHP for 24 hours to detect the gene expression and protein expression of apoptosis genes including Bax, Caspase-3 and Caspase-8.
Results:
The interference sequence of 3β-HSD gene inserted into lentivirus vector PLVX-shRNA2-puro is consistent with the designed sequence. 3β-HSD gene expression level in MCF-7 cells with 3β-HSD gene silencing was 77% lower than than that of control MCF-7 cells. 3β-HSD protein level in MCF-7 cells with 3β-HSD gene silencing was 74% lower than that of control MCF-7 cells. After DEHP treatment in MCF-7 cells with 3β-HSD gene silencing and control MCF-7 cells, qRT-PCR results showed that Bax gene expression levels increased by 28%-54%, Caspase-3 gene increased by 13%-49%, Caspase-8 gene increased by 21%-70% in MCF-7 cells when compared with the control group. Additionally, in the 3β-HSD gene silencing cells, Bax gene expression level decreased by 11%-28%, Caspase-3 gene expression decreased by 12%-23%, Caspase-8 gene expression decreased by 11%-34%, compared with the same treatment group of MCF-7 cells. Western blot results showed that Bax protein expression level increased by 28%-61%, Caspase-3 protein expression level increased by 40%-48%, Caspase-8 protein increased by 31%-84% in MCF-7 cells when compared with the control group. In 3β-HSD gene silencing cells, Bax protein expression level increased by 11%-27%, Caspase-3 protein increased by 21%-40%, Caspase-8 protein increased by 12%-25%, compared with the same treatment group of MCF-7 cells.
Conclusion
The stable 3β-HSD gene silencing cell line are successfully constructed in this study. DEHP can induce increased expression of apoptotic gene and protein. Silencing of 3β-HSD gene can inhibit the activation of apoptotic gene by DEHP in a certain degree.