1.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.
2.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.
3.A functional variant at miR-520a binding site in PIK3CA alters susceptibility to colorectal cancer in a Chinese Han population
Lifang DING ; Zao JIANG ; Qiaoyun CHEN ; Rong QIN ; Yue FANG ; Hao LI
China Oncology 2015;(3):184-189
Background and purpose: Increasing evidence has indicated that polymorphisms in the microRNA (miRNA, miR) binding site of target gene can alter the ability of miRNA and modulate the risk of cancer. We aimed to investigate the association between a miR-520a binding site single nucleotide polymorphism (SNP) rs141178472 in the PIK3CA 3’UTR and the risk of colorectal cancer in a Chinese Han population. Methods:The polymorphism rs141178472 was analyzed in a case-control study, including 386 colorectal cancer patients and 394 age-and sex-matched controls. The relationship between the polymorphism and the risk of colorectal cancer was examined by statistical methods. Results:Individuals carrying the rs141178472 CC genotype or C allele had an increased risk of developing colorectal cancer (CC vs TT, OR=1.716, 95%CI:1.084-2.716, P=0.022;C vs T, OR=1.258, 95%CI:1.021-1.551, P=0.033). Furthermore, the expression of PIK3CA was detected in the peripheral blood mononucleated cell of colorectal cancer patients, suggesting that mRNA levels of PIK3CA might be associated with SNP rs141178472. Conclusion:These ifndings provide evidence that a miR-520a binding site polymorphism rs141178472 in the PIK3CA 3’UTR may play crucial roles in the etiology of colorectal cancer.
4.Karyotypes analyses in 632 autism spectrum disorder trios
Tianlan LU ; Zhiliu WU ; Yanyan RUAN ; Ang MEIXIJIA ; Weihua YUE ; Jun LI ; Lifang WANG ; Dai ZHANG
Chinese Journal of Nervous and Mental Diseases 2016;42(3):150-155
Objective To detect chromosomal aberrations of autism spectrum disorder (ASD), we performed karyo?types analyses in 632 ASD trios and then investigated whether copy number variants and neurodevelopment related genes are present in the regions of chromosomal aberrations. Methods Karyotypes analyses were performed in 632 ASD trios (1896 individuals). In addition, we investigated whether there were pathogenic copy number variants located in the rele?vant regions of detected aberrant karyotypes by using the database of the International Standards for Cytogenomic Arrays (ISCA) and the Genomic Variation and Phenotype in Humans using Ensembl Resources (DECIPHER) for ASD patients. Results We detected aberrant results in 22 of 632 patients (3.48%) by karyotypes analyses. Of these 22 aberrant karyo?types, 5 were de novo (0.79%), including the duplication, the translocation, karyotypes of Turner syndrome and the addi?tional material with unknown origin. Seventeen children affected with autism had aberrant karyotypes inherited from one of their parents. By using the ISCA and the DECIPHER database, we found that several copy number variants with high pathogenicity were located in 1q25 and 3p24. Further, these copy number variants consisted of several genes related to neurodevelopment such as TNR, ASTN1, and NMNAT2. Conclusion There are a few de novo chromosomal aberrations in some patients affected with ASD. Copy number variants of several pathogenic neurodevelopmental related genes may exist in the regions of chromosomal aberrations. Karyotypes analyses may be applied to explore the genetic etiology in some patients affected with ASD.
5.Association Study of the Disrupted in Schizophrenia 1(DISC1)Gene Polymorphisms with Schizophrenia
Wei SUN ; Jun YAN ; Lifang WANG ; Mei QU ; Yanyan RUAN ; Tianlan LU ; Weihua YUE
Chinese Mental Health Journal 2002;0(08):-
T,P=0.008; Genotype:P=0.031)of DISC1 gene were significantly associated with schizophrenia.The haplotypes constructed by these two markers were significantly associated with schizophrenia,such as AT(?2=7.065,P=0.008,OR=1.42,95%CI=1.10~1.83)and GA(?2=6.009,P=0.014,OR=0.80,95%CI=0.68~0.96).When the subjects examined with the positive and negative syndrome scale(PANSS),the risk haplotype AT was not significantly correlated with positive,negative,excitement,depression and cognitive impairment factors of PANSS. Conclusion:These findings provide further evidence for DISC1 as a predisposing gene involved in schizophrenia in the Chinese Han Population.However,no positive association is found between DISC1 polymorphisms with schizophrenia clinical symptoms.
6.Estabishment and application of modified lung reperfusion injury animal model
Lifang WANG ; Xiaoming TIAN ; Qian PAN ; Yue LIU ; Tegeleqi BU ; Qing ZHANG ; Zhenrong ZHANG
Chinese Journal of Thoracic and Cardiovascular Surgery 2021;37(1):29-32
Objective:In this study, we established a reliable surgical procedure of lung ischaemia-reperfusion(IR) injury in rats. The research progress of different lung IR injury models and application value was also discussed.Methods:Twenty-eight adult SD rats were randomly divided into SHAM group and lung IR injury group(IR group), 14 rats in each group. In IR group, rats underwent tracheotomy under general anesthesia and received mechanical ventilation. Chest was opened in supine position, and pulmonary hilum was blocked for 30 minutes then the occlusion was removed. Samples were harvested after reperfusion for 45minutes. Rats in SHAM group received surgery and exposure of the right pulmonary artery, and experienced the same amount of time before the chest closed. Arterial blood gas was extracted postoperatively. Gross view of the lungs and pathological changes were observed, and the dry/wet ratio(W/D) was determined. Protein level of pro-inflammatory factors, markers in oxidative stress pathway, and endothelial functional markers in lung were tested by western blot analysis.Results:In IR group, there was pink foamy secretion in the airway, and the lungs exhibited signs of edema and congestion. In IR group, the alveolitis score was significantly increased, the W/D ratio was also increased, p38MAPK and NF-κB signaling pathways were activated, and the expression of TNF-α was significantly increased, while the expression of eNOS was significantly decreased.Conclusion:Left hilum clamping and bilateral reperfusion injury in lung is a practical animal model, it is a simple, low-cost and repeatable animal model for further studies. No microsurgical instruments were required during the procedure. Lung IR injury is characterized by oxidative stress response, inflammatory response and endothelial cell dysfunction.
7.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.
8.Establishment of a ICR mouse model of systemic C. albicans infection induced by oral inoculation
Yinzhu LUO ; Jinchun PAN ; Lifang HE ; Fangui MIN ; Qiuying YE ; Rui CHEN ; Yue WU ; Ren HUANG ; Yu ZHANG
Acta Laboratorium Animalis Scientia Sinica 2016;24(6):591-595
Objective To establish a mouse model of systemic C. albicans infection by oral inoculation of the pathogen and observe the proliferation and distribution of C. albicans in vivo tissues. Methods Male ICR mice(n=46) were used as the experiment group(n=40) and blank group (n=6). Cotton swabs with C. albicans were used to infect the mice (7 × 106 CFU/mL), and the blank group with saline. The mice of the experiment group were randomly divided into two groups:model group A for clinical assessment (n=20) and model group B for tissue fungal burden detection (n=20). Clinical score, survival and autopsy were carried out among the model group A. Five mice were randomly killed from the model group B at 3 d, 5 d and7 d after infection, respectively ( blank group killed 2 mice each time) . Microbial load tablet method was used to detect the tissue fungal burdens in different tissues, meanwhile samples of tongue, esophagus, stomach, liver, kidney, lung of infected mice were taken for pathological examination. Results White spot appeared on the surface of tongue since 3 d postinfection and increased with time and finally caused death. The mortality reached over 50% at 5 d. C. albicans was not only detected from the tongue (87?5%), stomach (87?5%), liver (54?5%), kidney (50?5%), lung (20%) and heart (4%), but also was microscopically seen mycelia proliferation in the tongue, stomach, liver, and kidney , yet not seen in the control group, showing that C. albicans caused disseminated systemic infection through mucosal infection in mice. Conclusions C. albicans can induce opportunistic systemic infection by breakthrough the mucosal immune barrier, so as to increase the infection to death.
9.Difference of Prenatal Ultrasound in the Diagnosis of Absence of Nasal Bone and Short Nasal Bone in Fetuses
Lei YANG ; Hezhou LI ; Lifang YUE ; Haiyan LU
Chinese Journal of Medical Imaging 2017;25(12):923-925
Purpose To explore the difference of prenatal ultrasound in the diagnosis of absence of nasal bone and short nasal bone in fetuses.Materials and Methods The clinical data of 23 131 pregnant women who received prenatal ultrasound system screening in the second trimester in the Third Affiliated Hospital of Zhengzhou University from August 2015 to July 2016 were analyzed retrospectively.The fetuses with absence of nasal bone and short nasal bone as well as the types of complicated malformation were recorded respectively,among whom those with chromosomal examination had their chromosomal results recorded.The fetuses were grouped for analysis based on absence of nasal bone,short nasal bone and whether they were complicated with malformation.Results Sixtynine fetuses had abnormal development of nasal bone,among whom 24 cases had absence of nasal bone (7 cases were complicated with malformation) and 45 cases had short nasal bone (13 cases were complicated with malformation),but there was no significant difference in the ratio of complicated malformation between fetuses with absence of nasal bone and those with short nasal bone (P>0.05).Of the 41 cases with analysis of chromosomal karyotypes,32 cases had single abnormal development of nasal bone (4 cases had chromosomal abnormality) and 9 cases had abnormal development complicated malformation of nasal bone (7 cases had chromosomal abnormality),and the occurrence rate of chromosomal abnormality in abnormal development complicated malformation of nasal bone was notably higher than that in single abnormal development of nasal bone (P<0.05).A total of 32 fetuses with single abnormal development of nasal bone received chromosomal examination,among whom 4 cases had chromosomal abnormality.Of the 32 fetuses,9 cases had single absence of nasal bone (one had chromosomal abnormality) and 23 cases had single short nasal bone (3 cases had chromosomal abnormality).However,there was no significant difference in the ratio of chromosomal abnormality between fetuses with single absence of nasal bone and those with single short nasal bone (P>0.05).Conclusion Abnormal development complicated with malformation of nasal bone is of great significance in the evaluation of chromosomes.And there is no significant difference in the ratios of complicated malformation and chromosomal abnormality between fetuses with absence of nasal bone and those with short nasal bone.
10.Research progress on fatigue in recipients of renal transplantation
Yue LI ; Yingzi MING ; Quan ZHUANG ; Xiaoxia WU ; Dongfang YIN ; Lifang LIU ; Jia LIU
Organ Transplantation 2020;11(2):311-
Renal transplantation is one of the most effective treatment methods for end-stage renal diseases. However, some recipients present with fatigue symptoms after renal transplantation. Fatigue not only affects the quality of life, but also reduces the compliance of recipients with immunosuppressive agents. To strengthen the attention of medical staff to the fatigue, make early diagnosis and deliver effective interventions for renal transplant recipients, the current situation, risk factors and intervention methods of fatigue in renal transplant recipients were reviewed in this article.