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.

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.

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.

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.

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.

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.

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.

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.

Objective:To systematically evaluate effects of auricular acupoint pressing on improving perimenopausal insomnia.Methods:The randomized controlled trials of auricular plaster therapy to improve perimenopausal insomnia was searched by computer in domestic and foreign databases, and the search time was from the establishment of each database to October 25, 2018. Two researchers independently screened the literature, extracted data and evaluated the risk of bias in the included literature and RevMan 5.3 analysis software was used for data analysis.Results:A total of 6 Chinese literatures were included. Meta-analysis results showed that there was no statistically significant difference in Pittsburgh Sleep Quality Index (PSQI) score reduction by auricular acupoint pressing compared with estazolam ( WMD=-0.06, 95% CI: -0.96-0.84, P=0.90) .Auricular acupoint pressing was better than Anshen Buxin capsule in reducing PSQI score ( WMD=-5.16, 95% CI: -7.37--2.95, P<0.000 01) . Auricular acupoint pressing could effectively improve perimenopausal insomnia, and the difference was statistically significant ( RR=1.19, 95% CI: 1.08-1.31, P=0.000 4) . Auricular acupoint pressing with beans combined with oral modified Huanglian Ejiao Decoction was better than oral modified Huanglian Ejiao Decoction alone in reducing Athens Insomnia Scale (AIS) scores ( WMD=-2.60, 95% CI: -4.42--0.78, P=0.005) . Compared with modified Chaihu Guizhi Longgu Muli Decoction treatment, auricular acupoint pressing combined with modified Chaihu Guizhi Longgu Muli Decoction could effectively reduce the modified Kupperman score ( WMD=-4.07, 95% CI: -6.36--1.78, P=0.000 5) . In terms of reducing the scores of the social support scale, there was no statistically significant difference between the auricular acupoint pressing with beans and estazolam ( WMD=-0.16, 95% CI: -1.26-0.94, P=0.78) . Conclusions:Auricular acupoint pressing has advantages in improving the total effective rate, reducing PSQI score and AIS score and improving Kupperman score. Limited by the quantity and quality of the included literatures, more large-sample high-quality studies with long-term follow-ups are needed for verification.

This paper reviews the literature about the common complications, evaluation tools and intervention measures of free flap repair for tongue cancer patients at home and abroad, summarizes the current research status, and points out that the nursing plan of free flap repair for tongue cancer patients needs to be further standardized, and needs to carry out evidence-based clinical research.