1.Effects of heat treatment and UVB radiation alone or in combination on the expression of heat shock protein 72 in human epidermal melanocytes
Lifang SHAO ; Guang ZHAO ; Jianrong NIU ; Rusong MENG ; Wenting SONG ; Yue ZHANG ; Yu CHENG ; Wenting YAN
Chinese Journal of Dermatology 2012;45(5):337-339
ObjectiveTo explore the effects of heat treatment and ultraviolet B (UVB) radiation alone or in combination on the expression of heat shock protein (HSP) 72 in human epidermal melanocytes.Methods Melanocytes were obtained from human foreskin,and subjected to primary culture.After 3 to 5 passages,the melanocytes were classified into 4 groups:control group (receiving no treatment),heat treatment group (treated with heat at 42 ℃ for 1 hour every day for 3 days),UVB group(irradiated with UVB at 50 mJ/cm2 daily for 3days),combination group(treated with heat at 42 ℃ for 1 hour followed by irradiation with UVB at 50 mJ/cm2daily for 3 days).After another 2- to 6-hour culture following the last treatment,melanocytes were collected and subjected to real time PCR and Western blot for the detection of HSP72 mRNA and protein expression,respectively.ResultsThe mRNA and protein expressions of HSP72 were significantly higher in the heat treatment group and combination group than in the control group (mRNA:6.584 ± 0.871 and 7.269 ± 0.454 vs.0.975 ± 0.089,both P < 0.001; protein:2.022 ± 0.058 and 2.080 ± 0.045 vs.0.532 ± 0.033,both P < 0.001 ),but was similar between the UVB group and control group (mRNA:0.832 ± 0.084 vs.0.975 ± 0.089,P > 0.05;protein:0.546±0.021 vs.0.532 ± 0.033,P > 0.05).The ANOVA of factorial design showed that neither heat treatment nor UVB irradiation had interaction effect on the mRNA or protein expression of HSP72 (F =2.106,1.399 respectively,both P < 0.05).ConclusionsHeat treatment can cause an increase in the expression of HSP72,which may enhance the function of melanocytes and protect melanocytes from UVB induced damage.
2.Effects of heat treatment on the activity of cultured human melanocytes irradiated by ultraviolet B in vitro
Wenting SONG ; Guang ZHAO ; Jianrong NIU ; Rusong MENG ; Yue ZHANG ; Lifang SHAO ; Yu CHENG ; Wenting YAN
Chinese Journal of Dermatology 2012;45(5):340-343
ObjectiveTo investigate the effect of heat treatment combined with narrow band ultraviolet B(NB-UVB) on cultured normal human melanocytes in vitro.MethodsMelanocytes were isolated from the foreskin of normal human,cullured in vitro,and irradiated with NB-UVB of different doses(20,30,50,70,90,120 and 180 mJ/cm2).Then,MTT assay was performed to evaluate the proliferation and activity of melanocytes to determine the optimal dose of UVB for the next experiment.Melanocytes were classified into 3 groups to be treated with heat at 42 ℃ for 1 hour (heat group),irradiated with UVB at 50 mJ/cm2 (UVB group),or irradiated with UVB at 50 mJ/cm2 followed by heat treatment at 42 ℃ for 1 hour (combination group),daily for 3 successive days; those receiving no treatment served as the control.After 24-hour culture following the last treatment,tyrosinase activity was evaluated with L-dopa as the substrate,melanin content was detected by NaOH assay,and cell cycle stages were determined by flow cytometry.ResultsNB-UVB irradiation decreased the viability of melanocytes in a dose-dependent manner,and the optimum dose of UVB was 50 mJ/cm2.The tyrosinase activity of melanocytes was 0.244 ± 0.018 and 0.310 ± 0.015 respectively in the UVB group and combination group,and increased by 3.8% (P < 0.05) and 31.9% (P < 0.05) respectively compared with the control group (0.235 ± 0.018); the melanin content was 0.201 ± 0.016 and 0.286 ± 0.019,respectively in the UVB group and combination group,and increased by 17.5% (P < 0.05 ) and 67.3% (P < 0.05) compared with the control group (0.171 ± 0.016).In comparison with the control group,the percentage of melanocytes in G1 phase was decreased by 23.94% in the UVB group(P< 0.05) and 33.51% in the combination group(P < 0.05),while that in S phase and G2 phase increased by 15.35% (P < 0.05 ) and 11.93% (P < 0.05),respectively in the UVB group,and 17.76% (P > 0.05) and 16.08% (P > 0.05),respectively in the heat group.ConclusionHeat treatment and NB-UVB can synergistically enhance the tyrosinase activity and accelerate melanogenesis,proliferation and differentiation,of melanocytes.
3.Association between leukoaraiosis and early neurological deterioration in patients with acute ischemic stroke
Wenting FU ; Changjuan SHAO ; Qing LI ; Sizhen XU
Chinese Journal of General Practitioners 2017;16(6):448-452
Objective To investigate the association between leukoaraiosis (LA) and early neurological deterioration (END) in patients with acute ischemic stroke.Methods Clinical data of 328 patients with acute ischemic stroke admitted in the hospital from January 2013 to January 2016 were retrospectively reviewed.According to the changes of National Institute of Health Stroke Scale (NIHSS) scores within 72 h after admission,88 patients (26.8%) were identified as END.Clinical manifestations,laboratory tests and radiographic findings were compared between END group and non-END group.ResultsUnivariate analysis indicated that age [(74.6±11.0) vs.(70.7±11.8) years,t=2.67,P=0.01],female sex [51.1% (45/ 88) vs.38.8%(93/240),χ2=4.05,P=0.04],initial NIHSS [M(Q1,Q3) 6(3,9) vs.3 (2,6),χ2=-4.38,P=0.00],systolic blood pressure [(155±28) vs.(149±20) mmHg(1 mmHg=0.133 kPa),t=2.04,P=0.04],responsible artery occlusion [18.2% (16/88) vs.8.3%(20/240),χ2=6.39,P=0.01],white cell count [(7.8±2.7) 109 vs.(7.1±2.2) 109,t=2.32,P=0.02],fasting blood glucose [(7.2±2.6) vs.(6.6±2.4) mmol/L,t=2.00,P<0.05] and C-reactive protein level [(24.5±27.1) vs.(14.6±23.2) g/L,t=3.25,P=0.00] were significantly different between END group and non-END group.After adjustment of confounding factors,LA in periventricular with Fazekas grade 2 (OR=2.309,95%CI: 1.070-4.984,P=0.03) and Fazekas grade 3 (OR=2.861,95%CI: 1.214-6.742,P=0.02) and LA in centrum semiovale with Fazekas grade 3 (OR=3.047,95%CI: 1.244-7.461,P=0.02) were independently associated with END.Conclusion Leukoaraiosis in periventricular group and centrum semiovale are associated with early neurological deterioration in patients with acute ischemic stroke.
4.Influences of infrared irradiation on human epidermal melanocytes
Yue ZHANG ; Guang ZHAO ; Rusong MENG ; Wenting SONG ; Lifang SHAO ; Jianrong NIU
Chinese Journal of Dermatology 2012;45(2):117-120
ObjectiveTo observe the increasing effect of infrared irradiation on tyrosinase activity and melanin content in cultured normal human epidermal melanocytes in vitro and to explore the optimal dose of infrared irradiation.MethodsEpidermal melanocytes were isolated from normal human foreskin tissue,and subjected to primary culture.Methyl thiazolyl tetrazolium(MTT) assay was performed to evaluate the effect of different doses(0,20,60,80,100,140,240,320 J/cm2)of infrared light on the proliferation of melanocytes and to select the optimal irradiation dose.Then,melanocytes were irradiated with infrared light at the optimal dose for 3 consecutive days followed by the determination of tyrosinase activity,melanin content,and cell cycle via dopa oxidation assay,NaOH solubilization method and flow cytometry,respectively.ResultsThe best intervention dose of infrared light was 80 J/cm2.The tyrosinase activity(A492 nm) and melanin content(A492 nm)were 0.3601 ± 0.0301 and 0.2748 ± 0.0243 respectively in melanocytes after irradiation with infrared light of 80 J/cm2 for 3 days,significantly higher than those in unirradiated melanocytes(0.3114 ± 0.0341,0.2325 ±0.0254,respectively,both P < 0.05),with an increase rate of 15.64% and 18.19% respectively.Cell cycle analysis revealed a decline in cell percentage in G1 phase(P < 0.01 ) but a concomitant increase in cell percentage in G2 and S phase (both P < 0.05) in irradiated melanocytes compared with unirradiated melanocytes.ConclusionsThe optimal dose of infrared light is 80 J/cm2 for the irradiation of melanocytes,and this dose of infrared light can increase melanin content,tyrosinase activity,differentiation and proliferation of melanocytes.
5.Clinical characteristics and prognosis of 44 patients with Nocardia infection
Xushuo XIE ; Qingluan YANG ; Jianming ZHENG ; Wenting ZENG ; Zengwei LIANG ; Lingyun SHAO ; Wenhong ZHANG
Chinese Journal of Infectious Diseases 2021;39(1):35-39
Objective:To investigate the clinical characteristics and prognosis of the patients with nocardiosis.Methods:From January 2013 to July 2019, 44 patients with nocardiosis in Department of Infectious Diseases, Huashan Hospital, Fudan University in Shanghai were enrolled, and their clinical data were retrospectively analyzed, including baseline characteristics, clinical manifestations, underlying diseases history of glucocorticoid therapy, laboratory data (blood routine examination, procalcitonin, C-reactive protein, lymphocytes subsets, etc.), imaging changes, bacterial strain identification, treatment regimens and outcomes. According to the locations of infection, patients were divided into pulmonary nocardiosis, extrapulmonary single-organ nocardiosis and disseminated nocardiosis. The Mann-Whitney U test was used for comparison between two groups, and the Kruskal-Wallis H test was used for comparison among multiple groups. Results:Among the 44 cases of nocardiosis, 14 cases were pulmonary nocardiosis, 17 cases were extrapulmonary single-organ nocardiosis (including nine cases with central nervous system infection, six cases with skin and soft tissue infection, one case with abdominal abscess and one case with urinary tract infection) and 13 cases were disseminated nocardiosis (including four cases with bloodstream infection, six cases with central nervous system and lung or skin and soft tissue infection, three cases of lung and skin and soft tissue infection). Thirty-four cases had underlying diseases, and 27 cases received glucocorticoid or immunosuppressant treatment. The main symptom of 11 patients in pulmonary nocardiosis group was productive cough, while that of the patients in other two groups was fever. Nocardia species were mainly Nocardia brasiliensis, Nocardia nova and Nocardia farcinicaia. The white blood cell counts and neutrophils proportion were normal or slightly increased in 42 cases, and the platelets were normal or slightly decreased in 41 cases. Erythrocyte sedimentation rate increased in 19 cases, procalcitonin increased in 21 cases, C-reactive protein increased in 34 cases, and ferritin increased in 18 cases. A total of 34 patients were tested for lymphocyte subsets, of which 15 had CD4 + T lymphocytes decreased, 14 had CD8 + T lymphocytes increased, seven had B lymphocytes increased, seven had B lymphocytes decreased, and eight had natural killer cells decreased. The hemoglobin of patients with pulmonary nocardiosis was higher than that of patients with extrapulmonary infection, and the difference was statistically significant ( U=0.095, P=0.025). The imaging manifestations were mainly abscess and inflammatory exudation. Forty cases were cured or improved, one case was still on treatment, and three cases died. Conclusions:The clinical manifestations of nocardiosis involving various organs are non-specific. Standardized treatment could reduce the mortality of nocardiosis.
6.A new method of extracting FECG by BSS of sparse signal.
Wenting SHAO ; Bin FANG ; Pu WANG ; Mingrong REN ; Yi SHEN
Journal of Biomedical Engineering 2009;26(6):1206-1210
The electrocardiograph signal presents the sparseness. In this paper, a new method of extracting the Fetal Electrocardiograph (FECG) is proposed, which uses the Blind Source Separation(BSS) of Sparse Signal. Both the Wavelet Analysis method and a new scheme named Vanish Circle are also involved in order to avoid the influence on the signal separation, because the ECG signals are not completed sparse signals. Some experiments show that the new method is effective.
Adult
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Algorithms
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Electrocardiography
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methods
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Female
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Fetal Heart
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physiology
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Fetal Monitoring
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methods
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Humans
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Pattern Recognition, Automated
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methods
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Pregnancy
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Signal Processing, Computer-Assisted
7.The mechanism underlying circHIPK3 promotes the migration and invasion of nasopharyngeal carcinoma cells 5 ⁃8F
Wenting Shao ; Lei Lv ; Jin Gao
Acta Universitatis Medicinalis Anhui 2023;58(8):1346-1353
Objective :
To explore the role and molecular mechanism of circHIPK3 in the migration and invasion of
nasopharyngeal carcinoma cells.
Methods :
Firstly , we searched public databases and analyzed the expression level of circHIPK3 in nasopharyngeal carcinoma tissues and non⁃cancerous tissues in Gene Expression Database (GEO) . Then , the circHIPK3 siRNA was synthesized and transfected into the NPC cell line 5 ⁃8F. After validate the efficiency of circHIPK3 knockdown , the proliferation , migration and invasion capacities of 5 ⁃8F cells were evaluated by CCK⁃8 , wound healing and transwell assay. Meanwhile , the mRNA and protein expression of downstream genes , including CDH1 , CDH2 , MMP2 , MMP9 and IL⁃6/STAT3 were determined by RT⁃qPCR and Western blot. Finally , transcriptome sequencing technology was used to analyze the significantly differentially expressed genes after the interference of circHIPK3 ⁃siRNA in 5 ⁃8F cells , in order to find out the key genes mediating the oncogenesis role of circHIPK3.
Results :
circHIPK3 expression significantly increased in nasopharyngeal carcinoma tissues compared with non⁃cancerous tissues ( P < 0. 05 ) . After siRNA⁃specific knockdown of circHIPK3 expression in 5 ⁃8F cells , the proliferation , scratch healing rate , and the number of migrated /invaded cells were significantly reduced (P < 0. 05 , P < 0. 01 , P < 0. 05 respectively) . Meanwhile , the mRNA and protein levels of CDH1 significantly increased (P < 0. 001) , but the CDH2 , MMP2 , MMP9 , IL⁃6/STAT3 mRNA and protein levels were significantly reduced (P < 0. 05) . Further analysis by RNA⁃seq showed that the expression of AL645922. 1 , SCO2 , AL136295. 1 and ISY1 ⁃RAB43 was significantly up⁃regulated( P < 0. 05 ) , and the expression of BIVM⁃ERCC5 , FAM47E⁃ST⁃BD1 , INO80B⁃WBP1 , NAIP , C15orf38⁃AP3S2 , BCL2L2⁃PABPN1 and SMIM11B was significantly down⁃regulated (P < 0. 05) in circHIPK3 ⁃siRNA transfected 5 ⁃8F cells.
Conclusion
circHIPK3 expression is significantly upregulated in nasopharyngeal carcinoma tissues. circHIPK3 promotes 5 ⁃8F cells proliferation , migration and invasion by regulating the expression of E ⁃Cadherin , N ⁃Cadherin , MMP2 , MMP9 and IL⁃6/STAT3. Genes such as AL645922.1 and BIVM⁃ERCC5 may play a key role in promoting the migration and invasion of nasopharyngeal carcinoma 5 ⁃8F cells mediated by circHIPK3.