Objective To observe the latency and amplitude characteristics of event-related potential P300 under different target stimu-lus. Methods During January to December, 2014, 35 patients with memory impairment received P300 test under the percentage of target stimuli of 0.10 and 0.25. The latency, amplitude and area under amplitude were compared. Results There was no significant difference in la-tency (t<0.256, P>0.800), but the amplitude and area under amplitude increased under 0.10 compared with those under 0.25 (t>4.259, P<0.05). Conclusion Less target stimulation may improve the patients' attention and increase amplitude of P300, but no more interference in la-tency, suggesting latency of P300 is more stable for assessment of cognitive function.

Objective:To explore the DNA methylation patterns and methylation differential genes of patients with coal-burning-borne endemic fluorosis, and to provide a basis for study of the pathogenesis of fluoride-induced body injury.Methods:A case-control study was conducted in Shuicheng County, Liupanshui, Guizhou Province, ten patients with severe fluorosis were selected as the fluorosis group in Douqing Township, where people burning high fluorine coal in open range all year round; and ten people without fluorosis phenotype were selected as the control group in Huaga Township, where firewood was the main fuel. Peripheral blood samples were collected from the two groups of people. Reduced representation bisulfite sequencing (RRBS) technique was used to detect the whole genome DNA methylation pattern ( n = 4) and DNA differentially methylated region (DMR), the DMR differential degree (log 2Ratio) and KEGG pathway enrichment analysis were used to screen the methylation differential genes, and real-time PCR was used to verify the mRNA expression levels of the candidate methylation differential genes( n = 10). Results:The methylation pattern analysis results showed that the methylation levels of all C bases in the genome DNA of the fluorosis group and the control group were (61.53 ± 0.59)% and (62.48 ± 1.53)%, respectively; among them, the methylated levels at CG sites were (63.75 ± 0.65)%, (64.36 ± 1.01)%, at CHG sites were (13.79 ± 0.72)%, (16.69 ± 4.06)%, and at CHH sites were (25.12 ± 1.72)%, (29.77 ± 3.97)%. Compared with the control group, patients in the fluorosis group had 1 000 DMR distributed on different autosomes; and the chromosome 19 was the most with 104 segments. There were 978 DMR-related genes, including 265 hypermethylation genes and 713 hypomethylation genes; KEGG pathway enrichment analysis showed that methylation differential genes were mainly involved in cell metabolism, cancers, and phosphatidylinositol 3-kinase-protein kinase B (PI3K-AKT) and other signaling pathways; combined with the differential degree of DMR, the hypermethylated succinate dehydrogenase complex flavoprotein subunit A pseudogene 3 (SDHAP3, log 2Ratio = 3.487) and hypomethylated nuclear factor κB inhibitor kinase regulatory subunit γ (IKBKG, log 2Ratio =-4.436) were selected as the candidate genes. There were statistically significant differences in the mRNA expression levels of SDHAP3 (0.54 ± 0.08, 1.00 ± 0.00) and IKBKG (1.32 ± 0.39, 1.00 ± 0.00) between fluorosis group and control group ( F = 22.94, 15.09, P < 0.01 or < 0.05). Conclusion:There are a large number of methylation differential genes in the genomes of patients with coal-burning-borne endemic fluorosis and controls, the hypermethylated SDHAP3 and hypomethylated IKBKG may be involved in fluoride induced body injury.

Chalcone is a common scaffold in natural products with optimal properties and biological activities.In this study, we designed and prepared eight new coumarin-chalcone derivatives (5a-5h), and confirmed their structures by 1H NMR and 13C NMR. Their in vitro antifungal activity combined with fluconazole (FLC) against drug-resistant Candida albicans was tested by microdilution method.The results indicated that most chalcone derivatives showed good antifungal activity against drug resistant Candida albicans with FLC, particularly with compound 5g displaying better antifungal activity (MIC50 = 5.60 μg/mL) than FLC (MIC50 = 200 μg/mL) when combined with FLC, so, these derivatives could be used as synergists of antifungal drugs.

Objective To investigate the effect of simulated microgravity on the proliferation and differentiation of the human megakaryocyte cells in vitro. Methods The fourth generation rotating cell culture system (RCCS-4) was used to generate the simulated microgravity environment. The cell viability was assessed by trypan blue staining method. The proliferation of cells was assessed by cell counting method and CCK8 method. The CD41+/CD61+ cells rate and the cells cycle were detected by flow cytometry. The expression levels of thrombopoietin receptor (c-mpl) and transcription factors were detected with RT-PCR. Results After 24, 48, 72 h, culture under simulated microgravity resulted in a significant decrease in the cell number , proliferative activity, cells in the G2/M phase and levels of c-mpl mRNA expression in comparison with that under the normal gravity (P < 0.05). After 48 h and 72 h culture, CD41+/CD61+ cells ratio decreased and RUNX-1 mRNA expression was down-regulated in cells of the group SMG compared with that of the group NG (P < 0.05). Conclusion Microgravity can inhibit the proliferation and differentiation of human megakaryocyte cells in vitro. The mechanism may be that TPO/c-mpl pathway was inhibited by down regulating the expression of c-mpl which transcriptional inhibition lead to.

Objective:To prepare nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR and to examine its cytotoxicity against EGFR positive tumor cells.Methods:By using molecular cloning strategy,prokaryotic expression construct of pET28a-BI7D12-PE38KDEL was generated which consisted of nanobody 7D12 targeting EGFR in the form of a divalent fused with PE38KDEL,a truncated form of pseudomonas exotoxin A via a flexible peptide(G4S)4,and then transformed into E.coli BL21(DE3).Protein expression was induced by adding IPTG,purified by Ni-affinity column chromatography,and verified by Western blot.The binding capacity of the resulted immunotoxin to EGFR-positive cells A549,HT29,MCF-7 and EGFR-negative cells CEM,Jurkat were determined by flow cytometry assay,and its cytotoxicity against the target cells was examined.Briefly,tumor cells were treated with different dosage of the immunotoxin,and the killing efficacy of BI7D12-PE38KDEL on these cells were assessed by WST-1 assay after 72 hours.Results:The SDS-PAGE and Western blot results showed the recombinant immunotoxin BI7D12-PE38KDEL was successfully prepared,and majority of them was expressed in soluble form.BI7D12-PE38KDEL could selectively bind to EGFR-positive cells of A549,HT29,and MCF-7.More importantly,the immunotoxin exhibited much more significant killing effect on these EGFR positive cells compared to the negative control group of CEM and Jurkat cells(P<0.01).Conclusion:In the current study,the nanobody-based immunotoxin BI7D12-PE38KDEL targeting EGFR was successfully prepared and exhibited a superior inhibition effect for the growth of EGFR-positive cells.

Objective In order to evaluate the possible effects of myeloid-derived suppressor cells (MDSCs) on graft versus host disease (aGVHD) development and clinical outcomes,this study systematically detected the dynamic changes of MDSCs accumulation in patients during the first 100 days after allogeneic hematopoietic stem cell transplantation (allo-HSCT).Methods Peripheral blood was obtained from 30 patients and 10 healthy volunteers with heparin anticoagulant tubes for 6 mL.For patients,peripheral blood was collected during the first 100 days after allo-HSCT and MDSCs levels were detected by flow cytometry.For measuring the serum concentrations of IL-6,IL-10,IL-1β,TNF-α,Arg-1,HO-1 and iNOS,samples were analyzed using ELISA kits.Results Patients developing aGVHD were infused with significantly less number of MDSCs [(39.94 ± 8.383) 106/kg]than in those not developing aGVHD [(209.0 ± 57.68) 106/kg,P =0.002 6];Patients developing aGVHD Ⅰ-Ⅱ and patients without aGVHD received significantly greater number of MDSCs [(61.96 ± 13.67) 106/kg and (209.0 ± 57.68) 106/kg] than in those developing aGVHD Ⅲ-Ⅳ [(20.37 ±4.304) 106/kg,P =0.013 9].After allo-HSCT,the mean percentage of MDSCs increased markedly in patients developing aGVHD [(7.725 ± 1.460)％] as compared with those not developing aGVHD [(3.423± 1.044)％,P =0.021 3].The high MDSCs group (＞53.712 × 106/kg) showed more favorable clinical outcomes than in the low MDSCs group (≤53.712 × 106/kg).The 2-year overall survival rate as 100％ in high MDSCs group,and 50％ in low MDSCs group (P =0.001 3).The cumulative incidence of 2-year relapse was 6.250％ and 29.252％ in high MDSCs group and low MDSCs group respectively (P =0.112 3).The cumulative incidence of NRM was significantly lower in high MDSCs group (0％) than in low MDSCs group (49.519％,P=0.001 8).MDSCs frequencies significantly increased in patients developing aGVHD after allo-HSCT.After allo-HSCT,the concentrations of IL-6,IL-10,TNF-α,Arg-1,iNOS and HO-1 were significantly elevated in patients developing aGVHD.Conclusion The number of MDSCs when engraftment may be used as a predictor for the development and severity of aGVHD.MDSCs might be considered as a potential new approach to regulate transplant rejection and achieve long-term survival.

Objective: To investigate the survival status and influencing factors of HIV/AIDS patients after receiving highly active antiretroviral therapy（HAART）in Shangluo,Shaanxi Province,so as to provide evidence for improving the effect of HAART. Methods: HIV/AIDS patients who received HAART for the first time in Shangluo from 2010 to 2018 were investigated. Life table method was used to analyze the survival rate,mortality rate and median survival time of the subjects. A proportional hazards model was used to analyze the influencing factors for the survival time of HIV/AIDS patients. Results: A total of 286 HIV/AIDS patients were collected,and 27 of them died of AIDS. After HAART,the 1-year,cumulative survival rates of less than 1 year,4 years and 8 years were 93.95%,89.71% and 88.39%,respectively. The results of multivariate proportional hazards regression analysis showed that the patients aged 30 years when first received HAART had higher risk of death than those aged 18-29 years（RR：4.208-24.095,95%CI：1.219-79.491）;patients with AIDS had higher risk of death than those with HIV（RR=38.590, 95%CI：15.451-96.382）;patients by homosexual transmission（RR=3.425,95%CI：1.385-8.470）and non-sexual transmission（RR=10.299,95%CI：3.602-29.446）had higher risk of death than those by heterosexual transmission;patients with baseline CD4+T lymphocytes number of 200/μL and more（RR：0.133-0.170,95%CI：0.048-0.604）had lower risk of death than those with less than 200/μL. Conclusions The survival rate of HIV/AIDS patients in Shangluo after receiving HAART is relatively high. Age at the start of treatment,course of disease,route of infection and the number of CD4+T lymphocytes at baseline are the influencing factors of survival time.

Objective To investigate the correlation between the serum ferritin levels and the post-stroke depression (PSD). Methods From July 2014 to October 2015, the inpatients with the first-ever acute ischemic stroke were colected consecutively. Chemiluminescence microparticle immune assay was used to measure the serum ferritin levels within 24 h after admission. Depressive symptoms were screened by using the 17-item Hamilton depression scale (HAMD-17) at 3 months after onset. In patients with a HAMD-17 score ≥7, the depression was further diagnosed according to The Diagnostic and Statistical Manual of Mental Disorders, 4th edition. Results A total of 200 patients with the first-ever acute ischemic stroke were enroled, 55 (27. 5% ) of them were diagnosed as PSD. There were significant differences in the body mass index (BMI), years of education, waist circumference, high sensitive-C-reactive protein, homocysteine, National Institutes of Health Stroke Scale score (at baseline, discharge, and day 90), mRs score (at discharge and day 90), BI (at discharge and day 90), and the proportions of widowed or solitary patients between the PSD group and the non-PSD group (al P < 0. 05 ). The serum ferritin level in the PSD group was significantly higher than that in the non-PSD group ( median [ interquartile range], 261. 90[142. 10-364. 90] μg/L vs. 164. 40[132. 50- 195. 10] μg/L; Z = - 4. 814, P < 0. 001 ). Multivariate logistic regression analysis adjusted for confounding factors showed that the baseline serum ferritin level >136. 375 μg/L was an independent risk factor for PSD (odds ratio 1. 041 per 1-quartile increase, 95%confidence interval 1. 009-1. 239; P = 0. 045). Conclusions The elevated baseline serum ferritin level is associated with PSD.

Objective To investigate the bacterial vaginosis(BV) and mycoplasma infection status in female patients with infertility.Methods The vaginal discharge samples were collected from 541 female patients with infertility in the gynecological department of the Hainan Provincial People's Hospital from June to December 2013.The BV detection,mycoplasma culture and drug susceptibility test were simultaneously conducted.Results The mycoplasma culture positive rate was 44.5％ (241/541),in which ureaplasma mycoplasma (UU) had the highest detection rate,there were 223 cases ofinfection with the positive rate of 41.2 ％,there were 5 cases of mycoplasma hominis(Mh) infection with the positive rate of 0.9 ％,13 cases were UU + Mh mixed positive with the positive rate of 2.4％.BV infection was in 325 cases with the positive rate of 60.1％ (325/541).Among 241 cases of mycoplasma positive,the BV detection rate was 70.1％ (169/241),while among 300 cases of mycoplasma negative,the BV detection rate was 52.0％ (156/300),the BV detection rate had statistical difference between the mycoplasma positive group and mycoplasma negative group,the difference was statistically significant (P＜0.05).Ofloxacin in fluoroquinolones had the highest drug resistance,the sensitivity was only 7.1％.Josamycin had the lowest drug resistance,accounting for 3.7 ％.Conclusion The female patients with infertility is closely correlated with BV and mycoplasma infection.Therefore BV and mycoplasma census should be strengthened to achieve early diagnosis and early treatment.Mycoplasma drug susceptibility test shows that doxycycline,minocycline and josamycin have the highest sensitivity,which can be used as the first selection drugs for clinical doctor.

Objective To observe the effects of Wutou decoction(WTD)on Anxiety and Depression induced by the L5 spinal nerve ligation (SNL) mice. Methods The ICR male mice were randomly divided into sham,SNL,high-dose,medium-dose and low-dose group of Wutou decoction,and Pregabalin group,with 24 in each group. Except the sham group, SNL models were prepared in the remaining groups of mice. After successful modeling, the high, medium, and low dosages were administered with 12.60, 6.30, 3.15 g/kg of Wutou decoction,and pregabalin group was administered with 0.25 g/kg of pregabalin,and the sham group and the model group was given an equal volume of normal saline. All the administration was once per day, continuously last for 21 days. On the 7/14/21thdays after the operation, the depressive and eanxiety behavior were evaluated by the tests of the sugar water consumption, the open field and forced swimming detection. In addition, the number of the positive cells expressing the extracellular signal-regulated kinase (p-ERK) in hippocampus were analyzed on Day 7/14/21 in each group by Immunohistochemistry and the expression of both p-ERK and postsynaptic density protein 95 (PSD95) in the hippocampus were analyzed on Day 21 by Western blot. Results At 7, 14, and 21thday after administration, compared with the model group, the paw withdrawal mechanical threshold of mice(0.76 ± 0.21 vs.0.05 ± 0.03),(0.93 ± 0.33 vs.0.05 ± 0.01),(1.12 ± 0.20 vs.0.04 ± 0.01)in the high dose group of Wutou decoction respectively increased(P<0.01),and sugar consumption(1.86 ± 0.44 g vs. 0.84± 0.23 g), (1.84 ± 0.10 g vs. 1.02 ± 0.18 g), (1.63 ± 0.31 g vs. 0.75 ± 0.23 g) significantly increased(P<0.01),and immobility time 4 min after swimming(128.90 ± 35.27 s vs.180.30 ± 21.81 s),(125.50 ± 23.07 s vs.195.30 ± 27.70 s),(169.50 ± 23.07 s vs.207.50 ± 7.46 s)significantly decreased(P<0.01),and the retention time in the central area of the open field(35.35 ± 7.61 s vs.13.90 ± 2.27 s),(26.26 ± 3.61 s vs.13.08 ± 1.98 s),(24.04 ± 6.57 s vs.10.62 ± 3.38 s)significantly increased(P<0.01).After 21 days,the number of p-ERK positive cells in the high-dose group of Wutou decoction(11.00 ± 4.89 vs.33.67 ± 7.35)was siginificantly lower than that of the model group,and the hippocampus p-ERK protein(0.15 ± 0.09 vs.0.54 ± 0.04)siginificantly decreased,and the expression of PSD95 protein(0.32 ± 0.04 vs.0.57 ± 0.09)siginificantly decreased(P<0.01). Conclusions The Wutou decoction achieved significant anti-depressant and anti-anxiety effects,and regulated p-ERK expression levels of hippocampus in the late stage of NP.