1.Effects of Candida albicans on the expression of tumor necrosis factor-α and activation of the intracellular signaling molecule p38MAPK in a human acute monocytic leukemia cell line THP-1
Zhimin DUAN ; Leilei DU ; Rong ZENG ; Yongnian SHEN ; Suquan HU ; Weida LIU ; Qing CHEN ; Min LI
Chinese Journal of Dermatology 2015;(8):535-538
Objective To investigate the effects of Candida albicans on the expression of tumor necrosis factor-α(TNF-α)and activation of the intracellular signaling molecule p38 mitogen-activated protein kinase(p38MAPK)in a human acute monocytic leukemia cell line THP-1. Methods Some THP-1 cells were divided into several groups in vitro: two C. albicans groups treated with 105 CFU/ml and 106 CFU/ml heat-killed C. albicans respectively, a lipopolysaccharide (LPS)group treated with 100 μg/L LPS, a blank control group treated with RPMI 1640 medium, two dexamethasone-inhibited groups pretreated with 40 μg/L dexamethasone for 30 minutes followed by treatment with 106 CFU/ml heat-killed C. albicans and LPS respectively. After treatment for 1, 3 and 6 hours, real-time fluorescence-based quantitative PCR was performed to measure TNF-α mRNA expression in THP-1 cells in the above groups. Enzyme-linked immunosorbent assay(ELISA)was conducted to determine the level of TNF-α protein in the supernatant of THP-1 cells treated with 106 CFU/ml heat-killed C. albicans, 100 μg/L LPS or RPMI 1640 medium(blank control group)for 24 hours. Western blot was performed to measure the protein expression of p38MAPK and phosphorylated p38MAPK in THP-1 cells after treatment with 106 CFU/ml heat-killed C. albicans or RPMI 1640 medium (blank control group)for 30 and 60 minutes. Statistical analysis was carried out by using two-way analysis of variance, one-way analysis of variance and the least significant difference(LSD)-t test. Results Significant differences were observed in the mRNA expression level of TNF-α among the C. albicans groups, LPS group and blank control group (F = 110.98, P < 0.001). The mRNA expression level of TNF-α in THP-1 cells increased over time in a time-dependent manner after C. albicans treatment, with significant differences among different time points (F = 701.680, P < 0.001). Compared with the blank control group, both 106-CFU/ml C. albicans group and LPS group showed a significant increase in TNF-α protein expression (6385.70 ± 533.99 ng/L and 3212.06 ± 353.00 ng/L vs. 147.10 ± 0.53 ng/L, P < 0.001 and 0.005, respectively). An obvious increase was observed in the expression level of phosphorylated p38MAPK protein, but no significant changes were noted in that of p38MAPK protein, in THP-1 cells treated with 106 CFU/ml C. albicans for 30 and 60 minutes compared with the blank control group. The mRNA expression level of TNF-α significantly decreased in dexamethasone-pretreated 106-CFU/ml C. albicans group and LPS group compared with those without dexamethasone pretreatment(3.77 ± 0.62 vs. 208.50 ± 10.50, 6.20 ± 1.93 vs. 161.35 ± 1.65, both P < 0.001). Conclusions Heat-killed C. albicans can induce the activation of p38MAPK in and secretion of TNF-α by human THP-1 cells, which then participate in the innate immune response against C. albicans.
2.Application and advantages of robot-assisted laparoscopic surgery in the treatment of high-risk prostate cancer
Tianyuan XU ; Shan ZHONG ; Xianjin WANG ; Leilei XIA ; Wei HE ; Zhoujun SHEN
Chinese Journal of Urology 2015;36(7):518-522
Objective To evaluate the feasibility and advantages of robot-assisted laparoscopic radical prostatectomy (RALRP) in treating high-risk prostate cancer.Methods From Mar.2010 to Dec.2014,69 men with high-risk prostate cancer (clinical stage ≥ T3a,serum PSA ≥ 20 μg/L or biopsy Gleason score ≥8) underwent surgical treatment at our center.RALRP was performed in 44 cases,with a mean age of (66.7 ± 8.6) yrs (range:50 ~ 82) and a mean baseline PSA of (23.1 ± 11.2) μg/L (range:3.6-48.8).Transperitoneal approach was adopted in all cases.Twenty-five patients were treated with ORP.The mean age was (64.3 ± 5.9) yrs (range:52-75) and baseline PSA was (21.7 ± 10.2) μg/L (range:5.7-41.3).Baseline clinical features,including age,initial PSA,biopsy Gleason score and clinical staging,were comparable between two cohorts.Surgical outcomes after RALRP were analyzed and compared between groups.Results All RALRP procedures were successfully performed with da Vinci robotic system and there was no open conversion.Mean operation duration were similarly (158 ± 47)min for RALRP group and (152± 42)min for ORP group,respectively.The mean estimated blood loss and postoperative length of hospital stay in RALRP group were (328 ± 254) ml and (8.4 ± 3.1) days,both of which were significant lower than those in ORP group,(674 ± 302) ml and (14.4 ± 3.7) days.Two cases of urine leak and two of lymphatic leak occurred after RALRP,and a statistically insignificantly higher complication rate was observed for the ORP group,in which four of urine leak and three of lymphatic leak happened.The positive surgical margin rates of two groups were 20.5% and 24%,respectively.After the mean follow-up of 25 (RALRP) and 27 months (ORP),continence rate was significantly higher in the RARLP group (93.2% vs.72.0%).PSA failure was observed in seven RALRP cases (15.9%) and four ORP cases (16.0%).Conclusion Compared with traditional ORP,RALRP shows significant advantages,including less blood loss,fewer complications,shorter length of stay,better postoperative continence and equivalent oncologic outcome when dealing with high-risk prostate cancer cases.Robot-assisted surgery is a safe and efficacious surgical modality in this setting.
3.Identification of three kinds of Plumeria flowers by DNA barcoding and HPLC specific chromatogram
Leilei ZHAO ; Xiaoxue YU ; Jie SHEN ; Xinjun XU
Journal of Pharmaceutical Analysis 2018;8(3):176-180
DNA barcoding and HPLC specific chromatogram were used to identify three kinds of Plumeria flowers respectively. DNAs extracted from the three Plumeria species were amplified by PCR with universal primers, and the psbA-trnH region was selected. All the amplified products were sequenced and the results were analyzed by MEGA 5.0. Chemometric methods including principal components analysis and hierarchical clustering analysis were conducted on the SAS 9.0 software to demonstrate the variability among samples. In conclusion, the psbA-trnH of all samples were successfully amplified from total DNA and sequenced. These three varieties of Plumeria can be differentiated by the psbA-trnH region and clustered into three groups respectively through building neighbor joining tree, which conformed to their germplasm origins. However, it was hard to distinguish them by HPLC specific chromatograms combined with chemometrics analysis. These indicated that DNA barcoding was a promising and reliable tool for the identification of three kinds of Plumeria flowers compared to HPLC specific chromatogram generally used. It could be treated as a powerful complementary method for traditional authentication, especially for those varieties which are difficult to be identified by conventional chromatography.
4.Construction and application of an intelligent injury monitoring system in Yinzhou District
LI Xiaoyong ; DUAN Leilei ; WU Yiqun ; ZHONG Jieming ; LIN Hongbo ; SHEN Peng ; ZHAO Lei
Journal of Preventive Medicine 2023;35(11):1009-1012
Injury monitoring is an important approach for injury control and is an important part of comprehensive disease monitoring. With the development of medical digitalization, an intelligent injury monitoring system has been created in Yinzhou District, Ningbo City, Zhejiang Province using artificial intelligence techniques based on Ningbo Municipal Health Information Platform and has been applied across the district since 2019. The manual card-reporting mode has been transformed to intelligent card-reporting mode in this system, which achieves functions of epidemiological analyses of the monitoring data, early warning of high-incidence injuries, classified management of injury and quality control of report cards. Nearly 300 thousand cards have been automatically reported since the system was online available since November 2022, and the epidemiological characteristics of injury were preliminarily understood, which provide data supports to early earning and interventions of injury. The intelligent injury monitoring system greatly improves the injury monitoring efficiency and card-reporting quality, and saves a large number of manpower and material resources, which provides a powerful technical support to widespread injury monitoring.
5.Effects of the yeast form of Sporothrix schenckii on activation of p38MAPK and expression of interleukin-6 in human THP-1 macrophage-like cells
Caixia LIU ; Leilei DU ; Zhimin DUAN ; Rong ZENG ; Yongnian SHEN ; Suquan HU ; Weida LIU ; Qing CHEN ; Min LI
Chinese Journal of Dermatology 2017;50(8):562-566
Objective To evaluate effects of the yeast form of Sporothrix schenckii on activation of p38 mitogen-activated protein kinase (p38MAPK) and expression of interleukin-6 (IL-6) in macrophagelike THP-1 cells,which were differentiated from the human acute monocytic leukemia cell line THP-1.Methods THP-1 macrophage-like cells were divided into 3 groups to be treated with the yeast form of Sporothrix schenckii at a concentration of 2 × 106 colony-forming units (CFU)/ml (yeast form group),100 mg/L curdlan (curdlan group) and RPMI 1640 medium (blank control group) respectively.Real-time fluorescence-based quantitative PCR was performed to measure the mRNA expression of IL-6 in THP-1 macrophage-like cells in the above 3 groups after 3-and 6-hour treatment separately,and enzyme-linked immunosorbent assay (ELISA) to detect the level of IL-6 in the culture supernatant of THP-1 macrophagelike cells after 24-hour treatment.Western blot analysis was conducted to determine the protein expression of p38MAPK and phosphorylated p38MAPK (p-p38MAPK) in the above 3 groups after 30-and 60-minute treatment separately.Other THP-1 macrophage-like cells were pretreated with 100 nmol/L dexamcthasonc (a p38MAPK inhibitor) for 30 minutes,and then were divided into 3 groups to be treated with the yeast form of Sporothrix schenckii,curdlan and RPMI 1640 medium respectively,and changes in the level of pp38MAPK and mRNA expression of IL-6 were also detected.Statistical analysis was carried out with SPSS19.0 software by using one-way or multi-way analysis of variance and least significant difference (LSD) test.Results Significant differences in the mRNA expression of IL-6 in THP-1 macrophage-like cells were observed among the yeast form group,curdlan group and blank control group (F =5 552.22,P <0.001) after 3-hour treatment (56.81 ± 7.36,26.69 ± 1.22 and 0.97 ± 0.05,respectively) and 6-hour treatment (378.03 ± 16.67,276.24 ± 39.13 and 1.02 ± 0.04,respectively).Additionally,the yeast form group showed significantly higher mRNA expression of IL-6 after 6-hour treatment than that after 3-hour treatment (q =16.74,P < 0.001).After 24-hour treatment,the level of IL-6 in the culture supernatant of THP-1 macrophage-like cells also significantly differed among the yeast form group,curdlan group and blank control group (59.96 ± 18.16 pg/L,91.01 ± 17.27 pg/L,5.50 ± 2.30 pg/L,respectively;F =26.62,P < 0.01),and was significantly higher in the yeast form group than in the blank control group (P < 0.01).After 30-and 60-minute treatment,the protein expression of p-p38MAPK was significantly higher in the yeast form group than in the blank control group (both P < 0.01).Moreover,the mRNA expression of IL-6 (4.46 ± 1.03 vs.493.52 ± 113.87,P < 0.001) and protein expression of p-p38MAPK (2.29 ± 0.37 vs.4.55 ±0.46,q =10.81,P < 0.01) were both significantly lower in the yeast form group with dexamethasone pretreatment than in that without dexamethasone pretreatment.Conclusion In vitro treatment with the yeast form of Sporothrix schenckii can enhance the expression of IL-6 in human THP-1 macrophage-like cells by activating the p38MAPK signaling pathway.
6.Clinical study of self-designed Yiqi Huoxue Huayu Decoction combined with western conventional therapy in the treatment of chronic heart failure complicated with atrial fibrillation
Lixiao WANG ; Xiaoli LI ; Haijun TANG ; Qiang PU ; Jingwei WANG ; Leilei XU ; Anming SHEN
International Journal of Traditional Chinese Medicine 2022;44(7):749-753
Objective:To observe the effects of self-designed Yiqi Huoxue Huayu Decoction on cardiac function and serum endothelin (ET-1) and matrix metalloproteinase-9 (MMP-9) levels in patients with chronic heart failure (GHF) complicated with atrial fibrillation.Methods:A total of 100 patients with GHF complicated with atrial fibrillation treated in our hospital from January 2019 to June 2021 were selected as the study subjects, and divided into experimental group and control group according to random number table method, with 50 patients in each group. Patients in both groups were given cardiotonic, diuretic, vasodilator and other conventional treatment, and patients in the experimental group added self-designed Yiqi Huoxue Huayu Decoction. TCM syndrome score, left ventricular end diastolic diameter (LVEDD), left ventricular end systolic diameter (LVESD) and left ventricular ejection fraction (LVEF), serum ET-1 and MMP-9 levels, clinical efficacy, safety and incidence of adverse reactions were observed and compared between 2 groups before and after treatment.Results:The total response rate was 82.0% (41/50) in the observation group and 62.0% (31/50) in the control group, and the difference was statistically significant ( χ 2=4.96, P=0.026). After treatment, the symptoms scores of palpitation, shortness of breath, fatigue, chest and hypochasm pain in the observation group were significantly significantly lower than those in the control group ( t=5.28, 5.29, 5.62, 5.42, P<0.01). After treatment, the LVEDD[(51.23±6.59)mm vs. (55.65±6.17)mm, t=3.46], LVESD[(43.10±4.76)mm vs. (45.99±5.31)mm, t=2.87], serum ET-1[(65.79±8.29)μg/L vs. (79.83±10.08)μg/L, t=7.61], MMP-9 [(175.86±24.81)ng/L vs. (189.49±26.13)ng/L, t=2.68] in experimental group were significantly lower than those in the control group ( P<0.05), while LVEF [(50.01±7.6)% vs. (46.25±6.96)%, t=2.57] was significantly higher than that of the control group ( P<0.05). There were no significant differences in GPT, GOT, UA, SCr levels between both groups before and after treatment ( P>0.05). There were no obvious adverse reactions betewwn both groups. Conclusion:The self-designed Yiqi Huoxue Huayu Decoction can relieve the clinical symptoms, improve the heart function and serum ET-1 and MMP-9 levels in patients with GHF complicated with atrial fibrillation safely.
7.Effects of early rehabilitation on the gross motor function of infants at high risk of infantile cerebral palsy
Yongfeng HONG ; Jun XU ; Xianshan SHEN ; Leilei JI ; Xiuli KAN ; Xueming LI ; Jianxian WU
Chinese Journal of Physical Medicine and Rehabilitation 2017;39(9):686-691
Objective To explore the effects of rehabilitation starting at different stages on the gross motor function of infants with high risk of cerebral palsy.Methods 214 neonates (117 boys and 97 girls) at least 14 days old and with a neonatal behavioral neurological assessment scale (NBNA) score of less than 35 were randomly divided into four groups.They received 2 to 3 weeks of rehabilitation in the hospital,followed by 4 to 6 weeks of family rehabilitation and then another 2 to 3 weeks of hospital rehabilitation.The three groups started this course of treatment at different times:beginning when the babies were not older than 3 months for group Ⅰ,3 to 6 months for group Ⅱ,6 to 12 months for group Ⅲ and 12 to 24 months for group Ⅳ.In each individual case the rehabilitation in hospital was replaced by family rehabilitation only when the baby's overall development was normal or almost normal.Another 63 infants with high risk of cerebral palsy composed group Ⅴ.They received only the family rehabilitation.The rehabilitation in hospital included taking neurotrophic drugs,receiving physical treatment,hand manipulation,kinesiotherapy,cognitive training,conductive education,and using orthoses.Family rehabilitation was implemented by the parents once or twice a day,for 35 to 45 minutes each time following detailed,individualized exercise plans made up jointly by the rehabilitation doctors and therapists.Moreover,a clinical checkup was required every 1 to 2 months.At 3,6,12 and 24 months old,all of the subjects' gross motor ability was quantified using the gross motor function measure (GMFM) scale.The hospital stays and the number of subjects diagnosed with cerebral palsy and motor retardation were recorded.Results At 3 months old,the average GMFM scores of the groups were 5.00± 1.89 for group Ⅰ,6.80± 1.55 for group Ⅱ,8.44±1.26 for group Ⅲ,11.10±1.72 for group Ⅳ and 12.70±1.64 for group Ⅴ,the controls.All of the intergroup differences were significant.At 6 months old the average GMFM scores of groups Ⅰ to Ⅳ were not significantly different,but all were significantly lower than the group Ⅳ average.At 12 months old the average GMFM scores of groups Ⅰ,Ⅱ and Ⅴ were significantly higher than the group Ⅲl and Ⅳ averages.At 24 months old the groups' average scores decreased consistently from Ⅰ to Ⅴ,with significant differences between one group and the next.Group Ⅲ had the longest hospital stays,followed by groups Ⅰ,Ⅱ and Ⅳ.Those inter-group differences were significant except for between groups Ⅰ and Ⅱ.The incidence of cerebral palsy in groups Ⅰ and Ⅱ (38.10% and 29.79%) was significantly higher than in group Ⅲ (21.62%),Ⅳ (12.00%) or Ⅴ (3.17%).Moreover,the incidence of motor retardation in groups Ⅰ and Ⅱ (7.14% and 10.64%) was also significantly lower than in group Ⅲ(56.76%),Ⅳ (68.00%) or Ⅴ (36.51%),but the differences between groups Ⅰ and Ⅱ and between groups Ⅲ and Ⅳ were not significant.The incidence of excellent gross motor performance ratings at 24mo decreased significantly from group Ⅰ to group Ⅴ,with all of the inter-group differences significant.Conclusion Early and formal rehabilitation is of great importance for infants with a high risk of cerebral palsy.The earlier formal rehabilitation starts,the better is its effect.
8.Relationship between domestic violence and posttraumatic stress disorder among women living in the communities of Wuhan city
Leilei WANG ; Xuening CHANG ; Senbei YANG ; Yukai DU ; Min SHEN
Chinese Journal of Epidemiology 2016;37(12):1574-1577
Objective To understand the prevalence of domestic violence and the correlation between domestic violence and posttraumatic stress disorder (PTSD) so as to improve the mental health status of women in Wuhan city of China.Methods A cross-sectional study was conducted in the community health service center in Wuhan.Participants were women who came to the center for gynecological examination.Data on general condition,social support,injury,epidemiological characteristics of domestic violence and PTSD were gathered.Chi-square,student-t and CochranArmitage trend tests were employed to compare the distribution of each characteristic variable between PTSD and non-PTSD group.Binary logistic regression was used to analyze the correlation between domestic violence and PTSD.Results A total of 1 015 women were effectively surveyed.The life-time incidence rates of events related to domestic,psychological,physical and sexual violence were 29.36%,28.28%,6.60% and 3.55%,respectively.The overall prevalence rate of PTSD was 4.73%.Women who suffered from domestic violence had higher risk of PTSD (OR=2.11,95% CI:1.04-4.29) than those who never experiencing domestic violence.Women who suffered from psychological or physical violence were 5.06 times more likely to suffer from PTSD (OR=5.06,95%CI:1.91-13.42).Conclusion A strong correlation between domestic violence and PTSD was seen.Victims who suffered from domestic violence should be provided with psychological counseling to reduce the risk of PTSD.
9. Potential false-positive rate among the 'asymptomatic infected individuals’ in close contacts of COVID-19 patients
Guihua ZHUANG ; Mingwang SHEN ; Lingxia ZENG ; Baibing MI ; Fangyao CHEN ; Wenjun LIU ; Leilei PEI ; Xin QI ; Chao LI
Chinese Journal of Epidemiology 2020;41(4):485-488
Objective:
As the prevention and control of COVID-19continues to advance, the active nucleic acid test screening in the close contacts of the patients has been carrying out in many parts of China. However, the false-positive rate of positive results in the screening has not been reported up to now. But to clearify the false-positive rate during screening is important in COVID-19 control and prevention.
Methods:
Point values and reasonable ranges of the indicators which impact the false-positive rate of positive results were estimated based on the information available to us at present. The false-positive rate of positive results in the active screening was deduced, and univariate and multivariate-probabilistic sensitivity analyses were performed to understand the robustness of the findings.
Results:
When the infection rate of the close contacts and the sensitivity and specificity of reported results were taken as the point estimates, the positive predictive value of the active screening was only 19.67%, in contrast, the false-positive rate of positive results was 80.33%. The multivariate-probabilistic sensitivity analysis results supported the base-case findings, with a 75% probability for the false-positive rate of positive results over 47%.
Conclusions
In the close contacts of COVID-19 patients, nearly half or even more of the 'asymptomatic infected individuals’ reported in the active nucleic acid test screening might be false positives.
10.Roles of Dectin-1 in phagocytosis of Candida albicans by macrophage-like cells derived from a human acute monocytic leukemia cell line THP-1
Zhimin DUAN ; Leilei DU ; Caixia LIU ; Rong ZENG ; Yongnian SHEN ; Suquan HU ; Weida LIU ; Qing CHEN ; Min LI
Chinese Journal of Dermatology 2018;51(6):425-428
Objective To investigate the roles of Dectin-1 in phagocytosis of Candida albicans (C.albicans) by macrophage-like cells derived from a human acute monocytic leukemia cell line THP-1.Methods THP-1 macrophage-like cells served as the target cells,and were transfected with small interfering RNA (siRNA) targeting Dectin-1 to down-regulate the expression of Dectin-1 receptor (siRNA-Dectin-1 group).THP-1 macrophage-like cells transfected with nonsense siRNA (siRNA-NC) served as a negative control group.After transfection,the THP-1 macrophage-like cells in the above 2 groups were cocultured with heat-killed C.albicans separately.And then,fluorescence microscopy was performed to count THP-1 macrophage-like cells phagocytosing C.albicans,and flow cytometry was used to determine the mean fluorescence intensity (MFI) of dihydrorhodamine (DHR)-123 fluorescent cells.Statistical analysis was done by one-way analysis of variance (ANOVA) and t test with the SPSS19.0 software.Results After transfection with siRNA-Dectin-1,the mRNA and protein expression of Dectin-1 significantly decreased in THP-1 macrophage-like cells (t =26.163,P < 0.001).After 1-,2-,4-hour co-culture of THP-1 macrophagelike cells with C.albicans,fluorescence microscopy showed that the phagocytosis rates of C.albicans by THP -1 macrophage-like cells were significantly lower in the siRNA-Dectin-1 group than in the negative control group (17.5% vs.22.1%,18.6% vs.24.3%,39.2% vs.59.1%,respectively,all P < 0.05),so were the percentage of THP-1 macrophage-like cells phagocytosing more than 3 C.albicans cells (2.2% vs.4.7%,2.5% vs.5.4%,5.1% vs.8.3%,respectively,all P < 0.05).After 30-minute,1-,2-and 4-hour co-culture of THP-1 macrophage-like cells with DHR-123-labelled C.albicans,flow cytometry showed that the MFI of C.albicans-phagocytosing cells was significantly lower in the siRNA-Dectin-1 group than in the negative control group (36.8 vs.45.7,54.3 vs.62.4,72.1 vs.84.9,93.6 vs.116.7,respectively,all P < 0.05).Conclusion Dectin-1 receptor plays an important role in the phagocytosis of C.albicans by macrophages.