1.shRNAs targeting high mobility group box-1 lead to inhibition of E-selectin expression via homeobox A9 in human umbilical vein endothelial cells
Xiaojuan ZHANG ; Lili JIAO ; Zhenggang LUAN ; Xiaochun MA
Chinese Critical Care Medicine 2015;(8):662-666
ObjectiveTo approach the regulatory mechanism of high mobility group box-1 (HMGB1) on the expression of E-selectin in human umbilical vein endothelial cell (HUVEC).Methods Homeobox A9 (HOXA9) siRNA was transfected to HUVEC at logarithmic phase, real-time fluorescence quantitative polymerase chain reaction (real-time qPCR) and Western Blot were used to determine the HOXA9 mRNA expression and protein expressions; a blank control group and a nonsilence negative control group were set. HUVEC stable transfected with pRNA-u6.1/Neo-HMGB1 shRNA plasmids (HUVEC with low-expression HMGB1) was obtained, and HOXA9 and E-selectin mRNA expressions were determined with real-time qPCR; a nonsilence transfection group served as the negative control. The HOXA9 siRNA was transfected to HUVEC with low-expression HMGB1 as co-transfection group, and the E-selectin expressions was determined with real-time qPCR; a HMGB1 shRNA group and a HOXA9 nonsilence group served as control.Results① HOXA9 mRNA (2-ΔΔCT) and protein expression (integralA value) in blank control group were 1.094±0.115 and 1.031±0.060. Compared with nonsilence transfection group, HOXA9 siRNA transfection group could significantly reduced mRNA and protein expression of HOXA9 [HOXA9 mRNA (2-ΔΔCT): 0.257±0.030 vs. 1.035±0.091,t = 14.010,P = 0.002; HOXA9 protein (integralA value): 0.278±0.042 vs. 0.975±0.014,t = 27.310, P = 0.002].② Compared with nonsilence transfection group, HMGB1 shRNA transfection could up-regulate HOXA9 mRNA expression in HUVEC (2-ΔΔCT: 2.519±0.278 vs. 0.856±0.063,t = 10.100,P = 0.001), also could down-regulate E-selectin mRNA expression (0.311±0.046 vs. 1.080±0.201,t = 7.415,P = 0.000).③ Compared with HOXA9 nonsilence group and HMGB1 shRNA group, HMGB1 shRNA and HOXA9 siRNA co-transfected HUVEC cells could significantly elevate E-selectin mRNA expression (2-ΔΔCT: 3.445±0.428 vs. 1.085±0.212, 1.004±0.104,t1 = 8.507, t2 = 9.603, bothP< 0.001).Conclusion HMGB1 may regulate E-selectin expression through the HOXA9 regulation in HUVEC.
2.Bacterial Distribution and Antimicrobial Resistance of Nosocomial Infection in Ningxia:Monitoring by Ningxia Hui Autonomous Region Nosocomial Infection Surveillance System
Baozhong YANG ; Qi ZHU ; Lili LUAN ; Yan WEN ; Xuexiang QI
Chinese Journal of Nosocomiology 2009;0(20):-
OBJECTIVE To observe bacterial distribution and drug resistance in Ningxia.METHODS The patients with nosocomial infection mornitored by Ningxia Hui Autonomous Region Nosocomial Infection Surveillance System from Jan 2003 to Dec 2007 were analyzed and summarized.RESULTS Amomg 3276 isolates,1752 strains(53.48%) were Gram-negative bacilli,1471 strains(44.9%) were Gram-positive cocci and,53(1.62%) fungi.The most common pathogens were Escherichia coli(909),Staphylococcus aureus(509),S.epidermidis(260),Enterococcus faecalis(258),and Pseudomonas aeruginosa(206).Most of them were multidrug resistant.Most strains of Gram-negative bacilli were highly susceptible to imipenem,while most strains of Gram-positive cocci were highly susceptible to vancomycin.CONCLUSIONS Most pathogens of nosocomial infection are multidrug resistant,the resistance detection of bacteria has an important significance to clinical treatment and infection control.
3.Screening of candidate molecules interacting with protein kinase Wee1B from human ovary cDNA library and its regulation effect
Chao LIU ; Lili REN ; Zhidong LUAN ; Zhichao MENG ; Yimeng LIU ; Jianying XIAO
Journal of Jilin University(Medicine Edition) 2016;42(5):866-871
Objective:To screen the new candidate molecules interacting with protein kinase Wee1B by yeast two hybrid system, and to analyze their interaction with Wee1B in the early stage of mouse fertilized eggs by bioinformatics.Methods:The plasmid pcDNA3.1/V5-His-TOPO-Wee1B wild type encoding mouse Wee1B gene was used as template to construct bait plasmid pGBKT7 Wee1B and the bait plasmid pGBKT7-Wee1B was transformed into yeast competent cells at SD/Trp (SDO),SD/Trp/X-α-Gal (SDO/X)and SD/Trp/X α Gal/AbA plates (SDO/X/A)plates to detect the toxicity and self-activation ability of yeast and its expression in yeast using Western blotting method.The yeast cells containing pGBKT7-Wee1B were fused with human ovary cDNA library, the yeast plasmid transformation of Escherichia coli positive clones were sequenced after identified by yeast transformation.BLAST analysis was carried out in GenBank,and its effect on the development of mouse fertilized eggs was deduced according to the gene annotation.Results:The double enzyme digestion analysis and sequencing analysis results showed that the pGBKT7-Wee1B bait plasmid was successfully constructed.The plasmid was transformed into the yeast,and there were no clones in the SDO/X/A plates.The pGBKT7-Wee1B and pGBKT7 empty vectors were transformed into the yeast,the bacteria were inoculated on the SDO plates,and the clones were uniformly grown on the two SDO plates.The positive clones were picked and expanded in culture,the protein was extracted and Western blotting showed that pGBKT7 Wee1B was expressed in the yeast.The bait plasmids were fused with human ovary cDNA library and the positive clones inserted into the fragment were identified by PCR. Nine proteins which interacted with Wee1B protein kinase were screened out by sequencing and blast analysis,and the proteins which could be closely related to the development of mouse oocytes and the development of fertilized eggs were analyzed by bioinformatics analysis.Conclusion:Using the yeast two hybrid system from human ovary cDNA library,nine interacting proteins with Wee1B protein kinase are screened and these screened proteins may regulate mouse oocyte maturation and early embryo development through interacting with Wee1B.
4.Infection Episode and Related Risk Factors in Continuous Hemodialysis Patients: A Survey
Qijun WAN ; Yongcheng HE ; Shaodong LUAN ; Lili HU ; Bin MA ; Ailian ZENG ; Yuhui XIE
Chinese Journal of Nosocomiology 2006;0(02):-
OBJECTIVE To investigate the infection episode and related risk factors in continuous hemodialysis patients. METHODS The relationship among infection and etiologies of infection,nutritional status,pathogens and causes of chronic renal failure(CRF) were retrospectively analyzed in 180 continuous hemodialysis patients. RESULTS Totally 113 times infections were observed among the 86 inpatients under continuous hemodialysis.The main infectious site in hemodialysis patients was lungs.Thirty eight times were positive in 50 times of etiologic detection,Gram-negative germ was the most common(60.3%).Hemoglobin and serum albumin decreased obviously in infectious patients.Diabetes and systemic lupus erythematosus patients were more susceptible to infection.The hepatitis virus infections rate in hemodialysis patients was relatively high. CONCLUSIONS There is higher infections rate in continuous hemodialysis patients.Diabetes and systemic lupus erythematosus patients are more susceptible to infection.Anemia,lower serum albumin,old age and bad compliance are the susceptible factors.
5.Study on Anti-stress Activity of Thymopentin Ethyl Ester
Qiang CHI ; Peng XU ; Lina DU ; Shan LA ; Zhongshan LUAN ; Junyang XU ; Lili WANG ; Zhi YANG ;
Journal of Medical Research 2006;0(11):-
Objective To investigate the anti - stress activity of thymopentin ethyl ester. Methods Anti - stress activity was evaluated by heat stress model and chronic uncontrolled stress model. Heat stress model;of 60 KM mice were divided into six groups. Except the controlled group, the other mice were maintained in 42℃ for 1h. Chronic uncontrolled stress model:of 60 KM mice were divided into six groups. Except the controlled group, the other mice were given three different stimulations once a day for continuous 21 d. The controlled group and model group were injected saline 0.2ml, and the three test groups were respectively injected thympentin ethyl ester at 2mg/kg,0.2mg/kg, 0.02mg/kg subcutaneously. The positive controlled mice were given thymopentin 0.2mg/kg subcutaneously. At the end of the experiment, plasma corticosteroid, IL -2 and SOD were determined according to the kit instructions. Results The activities of thymopentin ethyl ester in suppressed corticosteroid up - regulation and the elevated plasma IL - 2 and SOD level were more significant than those of thymopentin(P
6.The mechanism of Galectin-9 in the immune regulation of the active CD4+T cells
Hong LUAN ; Qian ZHANG ; Le WANG ; Miao ZHANG ; Yan CHEN ; Xiaoli XU ; Xingai LI ; Lili LIU ; Jin YUAN ; Yongman Lü
Chinese Journal of Microbiology and Immunology 2012;32(9):792-797
Objective To study the immune regulation of Galectin-9 on the active CD4+T cells and demonstrate the mechanisms.Methods Lymphocytes were harvested from wild-type C57BL/6 mouse,from which na(i)ve CD4+T cells were separated via MACS and then stimulated with anti-CD3 antibody(Ab) (2.5 μg/ml),anti-CD28 Ab(5 μg/ml) and IL-2(100 ng/ml) for 3 days.The active CD4+T cells were divided into 3 groups:Control group,Galectin-9 group and Galectin-9+α-lactose group.We detected the cell proliferation level by CFSE fluorescence intensity and then dynamically observed the cell morphological changes.The proportion of CD4+CD69+T cell,Th1,Th2 and Th17 cell was valued; Meanwhile,ELISA was used to detect the cytokine levels of IFN-γ,IL-4,IL-10,IL-12,IL-17A and TGF-β1 secreted by lymphocytes.Also Western blot was used to observe the changes of T cell differentiation regulatory protein such as T-bet,GATA-3 and ROR-γt.Results Compared with control group and Galectin-9+α-lactose group,in Galectin-9 group,the cell morphology began to change at 2 h.Moreover the proportions of CD4+CD69+ T cell,Th1 and Th17 cells decreased (P<0.05),but no significant differences in Th2 cells.The level of IFN-γ,IL-12,IL-17A and TGF-β1 from the supernatant decreased (P<0.05),while Th2-type cytokines IL-4 and IL-1O did not change.In addition,the expressions of T-bet and ROR-γt were significantly down-regulated (P<0.05).Conclusion Galectin-9 inhibited Th1 and Th17-type immune response,while had no effect on Th2-type immune response.The mechanism of the immune regulation may be related to affect the expression of Th1 and Th17 specific transcription factors at transcription level.
7.Research on the quality management mode of transitional care: Based on Three-dimensional QualityStructure Mode
Yuanyuan CHEN ; Lili CHEN ; Xiaorong LUAN ; Hailing YANG ; Ping WANG
Chinese Journal of Practical Nursing 2017;33(28):2214-2219
Objective To explore a reasonable transitional care quality management model, in order to standardize the management system of transitional care. Methods Using "three-dimensionalquality structure" mode as the theoretical framework, after reviewed relevant literatures, we established the rudiment of quality evaluation index system.Twenty-two experts were enrolled,and expert consultation was conducted using Delphimethod. The experts were evaluated by authority, positivity and degree of opinion coordination. Results The experts′authority coefficient was 0.846;the coefficient of determination was 0.891;the degree of familiarity was 0.80.The positive coefficients of the two rounds of consultation were 0.958,0.957. The evaluation system for transitional care consisted of 3 level- 1 indicators,12 level-2 indicators,and 64 level-3 indicators.The Kendall coefficients of concordance of the three level Indicators were 0.519, 0.525, and 0.432(P<0.05 for all). Conclusion The quality evaluation index system with With high reliability, providing standard reference for the work of quality management and nursing quality improvement.
8.Expression and clinical significance of SSTR2, ADAM-12 and FLI-1 in small cell lung cancer tissue
Meng WANG ; Juan DENG ; Xuejing LUAN ; Lili JIANG
Chinese Journal of Primary Medicine and Pharmacy 2023;30(4):601-606
Objective:To investigate the expression and clinical significance of somatostatin receptor 2 (SSTR2), a disintegrin and metalloproteinase-12 (ADAM-12), and friend leukemia virus integration-1 (FLI-1) in small cell lung cancer tissue.Methods:Eighty-two patients with small cell lung cancer who received treatment in Haiyang People's Hospital from January 2020 to January 2021 were included in this study. All patients underwent radical surgical resection. Small cell lung cancer tissues and adjacent tissues more than 2 cm from the edge of cancer tissues were harvested. The positive expression rates of SSTR2, ADAM-12, and FLI-1 in cancer tissues and adjacent tissues were determined by immunohistochemistry. The relationship between SSTR2, ADAM-12, FLI-1, and clinical characteristics were analyzed. The 1-year survival rate of patients with small cell lung cancer was calculated.Results:The positive rates of SSTR2, ADAM-12, and FLI-1 in small cell lung cancer tissue were 79.27% (65/82), 76.83% (63/82), and 78.05% (64/82), respectively, which were significantly higher than 19.51% (16/82), 17.07% (14/82), 20.73% (17/82) in the adjacent tissue ( χ2 = 58.57, 58.78, 53.90, all P < 0.05). SSTR2, ADAM-12, and FLI-1 were positively associated with lymph node metastasis, clinical stage, tissue invasion, tumor size, and histological grade (all P < 0.05). After controlling for gender, age, and others, SSTR2, ADAM-12, and FLI-1 were associated with lymph node metastasis, clinical stage, tissue invasion, tumor size, and histological grade (all P < 0.05). All patients were followed up for 1 year. Six patients were lost to follow-up. The 1-year survival rate of 76 patients with small cell lung cancer was 67.11% (51/76). The survival rate of patients with positive SSTR2, ADAM-12, and FLI-1 expression were lower than that of patients with negative SSTR2, ADAM-12, and FLI-1 expression ( χ2 = 3.93, 6.43, 7.52, all P < 0.05). Conclusion:SSTR2, ADAM-12, and FLI-1 are highly expressed in small cell lung cancer tissue. Combined detection of SSTR2, ADAM-12, and FLI-1 is conducive to the prognosis and evaluation of small cell lung cancer in patients. This study is innovative and scientific.
9.The accuracy analysis of fully-guided mucosa-supported template based computer-assisted implant surgery
LUAN Lili ; XIAO Huijuan ; LIU Hui ; HAN Xiaopeng ; YU Hui ; LIU Zhonghao
Journal of Prevention and Treatment for Stomatological Diseases 2017;25(9):575-581
Objective :
The purpose of this study is to determine the accuracy of implant placement in the edentulous jaw using computer planning and fully-guided mucosa-supported surgical template by Simplant software.
Methods:
63 implants were placed in 9 patients (11 fully edentulous jaws), 26 implants were placed in upper edentulous jaw and 37 implants in lower edentulous jaw. Preoperatively, first, a cone beam CT was required for patients with radiographic template and radiographic template respectively. Therefore, the data of CBCT was inputted in Simplant software by DICOM format, followed by virtual implant planning. Hereafter, a mucosa-supported surgical template was designed by dentist and made by Masterilise company to allow implant placement using the template as a guide. To investigate the accuracy of implant placement, a postoperative CBCT scan was obtained and matched to the preoperative scan. The accuracy of implant placement was validated three-dimensionally including divations of implant shoulder, apical point, axial angulation and depth.
Results:
The survival rate of 63 implamts for a 6 month to 10 year observation period was 100%. The mean divation of implant shoulder was 0.73 ± 0.53 mm, implant apical point was 1.15 ± 0.62 mm, implant depth was 0.95 ± 0.64 mm and implant axial angulationwas 4.10 ° ± 3.23°.
Conclusion
Divation between virtual and actual implant was existed and it should be considered preoperatively when virtual implant was planned to avoid injuring anatomic structure and keeping surgery safely. Correct manipulation during implant operation are helpful to decrease the divation of implant placement.
10.Combating COVID-19 with integrated traditional Chinese and Western medicine in China.
Liqiang NI ; Lili CHEN ; Xia HUANG ; Chouping HAN ; Jianrong XU ; Hong ZHANG ; Xin LUAN ; Yongfang ZHAO ; Jianguang XU ; Weian YUAN ; Hongzhuan CHEN
Acta Pharmaceutica Sinica B 2020;10(7):1149-1162
COVID-19, an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has spread throughout the world. China has achieved rapid containment of this highly infectious disease following the principles of early detection, early quarantine and early treatment with integrated traditional Chinese and Western medicine. The inclusion of traditional Chinese medicine (TCM) in the Chinese protocol is based on its successful historic experience in fighting against pestilence. Current findings have shown that the Chinese medicine can reduce the incidence of severe or critical events, improve clinical recovery and help alleviate symptoms such as cough or fever. To date there are over 133 ongoing registered clinical studies on TCM/integrated traditional Chinese and Western medicine. The three Chinese patent medicines (/ (Forsythiae and Honeysuckle Flower Pestilence-Clearing Granules/Capsules), (Honeysuckle Flower Cold-Relieving Granules) and (Stasis-Resolving & Toxin-Removing) were officially approved by the National Medical Products Administration to list COVID-19 as an additional indication. The pharmacological studies have suggested that Chinese medicine is effective for COVID-19 probably through its host-directed regulation and certain antiviral effects.