Objective To observe the effect of Danshen in the treatment of severe hand-foot-and-mouth disease and investigate its mechanism.Methods One hundred and forty cases of children with severe hand foot and mouth diseases who hospitalized the intensive care unit were enrolled in the study from February 2012 to July 2014.The children were randomly divided into a control group and an observation group,with 70 cases in each group.The control group was given antiviral to reduce the intracranial pressure and intravenous immunoglobulin and the observation group was given Danshen in addition to the control group.The levels of neuron specific enolase (NSE)、S-100βprotein、TNF-α and IL-6 were detected at admission and checked again seven days after treatment.The changes of the above indicators and the clinical curative effect were compared before and after therapy.Results The total effective rate of the control group and the observation group was 80.0% and 92.9% respectively,showing statistical significance(P<0.05). After treatment, observation group of serum NSE (9.42 ± 2.56μg/Lvs. 11.71 ± 3.21μg/L,t=2.159), S-100β (177.34 ± 87.28 ng/Lvs. 286.14 ± 159.69 ng/L, t=2.315), TNF-α (15.98 ± 4.35 ng/Lvs. 23.17 ± 4.80 ng/L, t=4.297), IL-6 (41.72 ± 6.64 ng/Lvs. 52.05 ± 9.33 ng/L,t=3.492) , the level of were lower than those in the control group(P<0.05 orP<0.01). In the observation group, serum. The fever clearance time、the disappearance time of rash and the hospitalization time in the observation group (3.55 ± 1.02 d vs.4.55 ± 1.09 d, 7.14 ± 1.04 d) were shorter than those in the control group (4.46 ± 0.97 d vs.5.88 ± 1.44 d, 8.68 ± 1.06 d;t=5.409, 6.161, 8.677 respectively, P<0.01).Conclusions On the basis of conventional therapy,Danshen can effectively alleviate the systemic inflammatory response in children with severe hand foot and mouth diseases, reduce brain damage and improve the clinical efficacy.

Objective To investigate the effects of nephroblastoma overexpressed (NOV) on proliferation,adhesion,migration and invasion of remal cell curcinomai (RCC) cells. Methods We constructed a NOV expression plasmid and transfected the plasmid into RCC cell line 786-O and analyzed the effects of NOV expression on proliferation,adhesion,migration and invasion of RCC cells by growth curve assay,WST-1 assay,cell adhesion assay,matrigel invasion assay and transwell migration assay. Results The stable NOV transfected 786-O cells (786-O-NOV) showed decreased growth rate,at 48 h and 72 h,the proliferation activities of 786-O-NOV cells were inhibited by 29.14％ and 32.46％ the proliferation activities of empty vector cells were inhibited by 9.25％ and - 8.16％,respectively,compared to 786-O cells (P ＜0.05); while the 786-O cells transfected with empty vector (786-O-mock) had no difference with 786-Ocells.Adhesion assay indicated significantly increased adhesion of 786-O-NOV cells to fibronectin (0.26 ±0.03) and laminin (0.28 ±0.04),compared to 786-O cells (0.15 ±0.01,0.12±0.10) and 786-O-mock cells (0.14 ±0.02,0.13 ± 0.08).Invasion assay displayed that the numbers of cells penetrated through matrigel membrane were significantly higher in 786-O-NOV cells (240.25 ± 23.12) compared to 786-O cells ( 56.16 ± 6.25 ) and 786-O-mock cells ( 50.28 ± 7.13 ).Migration assay displayed that the numbers of cells passed through polycarbonate filters were significantly higher in 786-O-NOV cells (267.25 ± 20.94) compared to 786-O cells ( 66.10 ± 5.68 ) and 786-O-mock cells ( 56.28 ± 4.11 ).Conclusion NOV exhibits anti-proliferative effects on RCC cells; however,it promotes adhesion,migration and invasion of RCC cells.

Objective: To determine the value of telomerase activity in urine exfoliated cells as clinical indicator of tumor presence, stage, and recurrence. Methods: The techniques of TRAP-PCR and TRAP-sliver staining were employed to detect telomerase activity in 73 patients with TCC before operation (study group) and 20 benign urothelial cancer patients (control group) and 21 normal individuals (normal group). Cytologic results were obtained simultaneously. The bladder tumor specimens were obtained from 73 patients during operation, and histologically evaluated for tumor content and grade. Results: Positive rate of telomerase activity detection in TCC patients (80.8%,59 of 73) was significantly higher than that of cytological examination (20.5%, 15 of 73). Positive telomerase activity was not found in 17 of 20 in control group and none was found in normal group. Analysis of the distribution of abnormalities with tumor stage revealed greater detection of high pathological stage (T 2-T 4) (89.7%, 26 of 29) compared with low stage (Tis-T 1) (75%, 33 of 44). Conclusion: Detection of telomerase activity in urine exfoliated cells may be a sensitive and effective marker in the diagnosis of TCC.

Objective To determine the value of detection of H ras oncogene mutation in urine exfoliated cells as clinical indicator of tumor presence, recurrence and stage.Methods Point mutation at codon 12 of H ras gene was assayed by polymerase chain reaction followed by analysis of single strand conformation polymorphism in urine exfoliated cells from 48 patients with transitional cell carcinoma before operation and 28 patients with non urothelial cancer or normal individuals. The mutation was further confirmed by dideoxy mediated chain termination method of DNA sequencing. Cytology analysis was carried out simultaneously. Bladder tumor specimens were obtained from 48 patients during operation, and histologically elevated for tumor content and grading.Results 48%(23 of 48) of the patients were detected by their aberrant band in SSCP. All aberrant bands displayed a mutant H ras sequence, where 15% (7 of 48) of the patients displayed, apositive cytological analysis. Analysis of abnormalities with tumor stage revealed that the greater detection of high pathological stage (Ⅲ Ⅳ) compared with low stage (Ⅰ Ⅱ) was related to the recurrence of transitional cell carcinoma.Conclusion Our results suggest that the detection of H ras mutations may be of clinical value in the detection of TCC.

Objective:To investigate the value of tumor hemodynamics and morphological features from conventional dynamic contrast-enhanced MRI (DCE-MRI) scan before surgery in predicting postoperative recurrence time in breast cancer patients.Methods:A retrospective analysis of 58 patients with breast cancer who had recurred after operation from November 2012 to December 2014 in Liaoning Cancer Hospital was performed. According to the recurrence time, the patients were divided into early recurrence group (≤2 years after surgery, 33 cases) and late recurrence group (>2 years after surgery, 25 cases). All patients underwent routine DCE-MRI scans before surgery, and hemodynamic features of the three-dimensional volume of the tumor and the morphological and textural features of the tumor in each phase were extracted by computer. The counts and measurement data of patients in early recurrence group and late recurrence group were compared by Fisher′s exact probability method and Mann-Whitney U test, and receiver operating characteristic (ROC) curves were drawn. The multivariate logistic regression was used to calculate the combined efficacy in predicting early recurrence and late recurrence. Kaplan-Meier method was used to analyze the survival prognosis, and Log-Rank test was used to compare the differences in survival curves between groups. Results:There was no significant difference in background parenchymal enhancement, lesion margin, lesion internal enhancement characteristics, lesion morphology, time-signal intensity curve type and the degree of whole-breast vascularity increase between early recurrence and late recurrence groups ( P>0.05).There were significant differences in the maximum concentration of contrast (Max Conc), the area under the time signal curve (AUC) and the maximum slope value of the time signal curve (Max Slope) ( P<0.05). Comparative analysis of the radiomics parameters of 8 phases DCE-MRI found that the sphericity of morphological characteristic parameters in the phase 3 was statistically different between the early recurrence and late recurrence groups ( P=0.03). Area under the ROC curve of AUC, Max Conc, Max Slope and parameter sphericity of phase 3 morphological characteristics for predicting early and late recurrence were 0.664, 0.659, 0.684 and 0.670, respectively. The area under the ROC combined with the above four parameters for prediction was 0.765, with a specificity of 63.6% and a sensitivity of 84.0%; the predictive efficacy was higher than that of univariate. Fifty-eight patients were followed up for 17 to 64 months with a median follow-up of 47 months. The disease-free survival and overall survival in the early recurrence group were significantly lower than those in the late recurrence group, and the difference was statistically significant ( P<0.05). Conclusion:It is of certain value to predict the postoperative recurrence time of breast cancer based on the tumor hemodynamic characteristics combined with morphological characteristics from preoperative non-invasive conventional DCE-MRI.

Objective: To establish a liquid chromatography tandem mass spectrometry method to determine the cortisol in saliva. Methods: Take 0.5 ml saliva sample into a 2 ml centrifuge tube, add excess sodium chloride and 1ml acetonitrile to it, then vortex for 3 min, centrifuge for 10 min at 15 000 r/min, and take 800 μl of the upper layer to another centrifuge tube. Finally, the sample was concentrated by a vacuum concentrator and brought to 200 μl with the initial mobile phase. Then, the sample was analyzed by liquid chromatography tandem mass spectrometry. The target compound was quantified by external standard curve method. Results: The linear range of the method was 0.02-5.00 ng/ml, r=0.999 9, the method limit of the detection was 0.002 ng/ml, the method limit of quantitative was 0.02 ng/ml, and the spiked recoveries were 89.60%-98.60%. The intra-assay precision was 1.90%-3.30%, and the inter-assay precision was 4.20%-9.00%; samples could be stored at -20 °C for at least 14 days. The determination of cortisol could not be interfered by other endogenous substances in the sample. Conclusion The method is simple in pretreatment, high sensitivity, good reproducibility and good recovery, and it is suitable for the quantitative analysis of cortisol in saliva for normal and occupationally stressed populations.

Background Gene sequencing industry is an emerging innovation-driven industry. Employees have high requirements for independent learning and innovation ability and face great professional pressure. Objective To understand the occupational stress, depression, and sleep of gene sequencing enterprise employees and to analyze the effect of occupational stress on depression and sleep. Methods From November to December 2021, occupational stress, depression, and sleep conditions of 469 workers from 34 enterprises in gene sequencing industry were surveyed by Core Occupational Stress Scale (COSS), Patient Health Questionnaire 9 (PHQ-9), and Pittsburgh Sleep Quality Index (PSQI). A total of 427 valid questionnaires were recovered with a questionnaire valid response rate of 91.04%. The relationship of occupational stress with depression or sleep was analyzed by logistic regression. Results The rates of occupational stress, depression, and sleep disorder were 27.40%, 33.50%, and 28.10%, respectively. Significant difference were found in the rates of depression and sleep disorder in different occupational stress groups (P<0.05). The results of logistic regression analysis showed that, for every 1 increase in social support score, the risk of depression increased by 1.206 (95%CI: 1.117−1.304), and the risk of sleep disorder increased by 1.143 (95%CI: 1.059−1.233). For every 1 increase in organization and reward score, the risk of developing depression increased by 1.082 (95%CI: 1.017−1.151). Mild, moderate, and severe occupational stress were all associated with a higher risk of depression in reference to no occupational stress (OR=2.535, 95%CI: 1.465−4.386; OR=3.774, 95%CI: 1.809−7.870; OR=3.823, 95%CI: 1.486−9.837). Severe occupational stress was associated with a higher risk of sleep disorder in reference to no occupational stress (OR=3.141, 95%CI: 1.233−8.006). Conclusion Occupational stress among employees in the gene sequencing industry can increase the risks of depression and sleep disorder. Enterprises need to take intervention measures and pay attention to prevention and treatment.

Objective: To develop a method using ultra-high performance liquid chromatography-triple quadrupole mass spectrometry to determine the urinary metabolites of benzene, toluene and xylene. The selected metabolites are S-phenylmercapturic acid (S-PMA) , trans, trans-muconic acid (t, t-MA) , 8-hydroxy-2 deoxyguanosine (8-OHdG) , hippuric acid (HA) , 2-methylhippuric acid (2-MHA) , 3-methylhippuric acid (3-MHA) and 4-methylhippuric acid (4-MHA) . Methods: The urine sample was pretreated using methanol to precipitate the proteins. HSS T3 chromatographic column was used to separate the metabolites. The mass spectrometric acquisition was carried out using multiple reaction monitoring (MRM) after ionization with ESI source. External standard method was used for quantification. Results: All the standard curves showed good linear relation, and r of the seven metabolites was all above 0.999. The detection limits and quantitative limits of the seven metabolites were 0.01-500 ng/ml and 0.02-1 000 ng/ml (based on the actual dilution ratio) , respectively. The average spiked recoveries of four loadings ranged from 85.8% to 109.9%. The intra-day and inter-day precisions were 0.2%-4.5% and 0.6%-9.5%, respectively. The samples can be kept for at least 14 days at both 4 ℃ and -20 ℃. Conclusion This method is simple, rapid and highly sensitive with low cost, and its accuracy, precision and stability can meet the daily test requirements. It can be applied for the determination of urinary S-PMA, t, t-MA, 8-OHdG, HA, 2-MHA, 3-MHA and 4-MHA for the occupational population exposed to benzene, toluene and xylene.

ObjectiveTo analyze the epidemic characteristics of foodborne disease outbreaks in Mengla County， Yunnan Province， so as to provide basis for formulating corresponding prevention and control measures. MethodsThe data of foodborne disease outbreaks in Mengla County， Yunnan Province from 2017 to 2021 in the national "foodborne disease outbreak monitoring system" were collected. The time， population， region， place， pathogenic factors and inducing links of the events were statistically analyzed. ResultsFrom 2017 to 2021， a total of 68 incidents were reported， including 526 cases and 5 deaths. The total incidence rate was 23.40% and the case fatality rate was 0.95%. The peak period was from May to July， with the largest number of reported events in July. The reporting areas were mainly Mengla Town （35.29%）， Guanlei Town （11.76%） and Mengpeng Town （10.29%）. The main place of the incident was family （76.47%）. The main pathogenic factors were plant toxins （79.41%）， mainly through the ingestion of wild mushrooms （68.52%）， improper processing of green beans （12.96%） and aconitum （9.26%）. ConclusionAccording to the epidemiological characteristics of Mengla County， we should focus on the public education of health knowledge in key areas and places， especially on the identification of wild mushrooms， to prevent accidental eating. At the same time， we should strengthen the monitoring and risk assessment of foodborne diseases， find potential risks as soon as possible， issue early warning and forecast in time， and constantly improve the clinical treatment ability of poisoned patients.

Developing new therapeutic agents for cancer immunotherapy is highly demanding due to the low response ratio of PD-1/PD-L1 blockade in cancer patients. Here, we discovered that the novel immune checkpoint VISTA is highly expressed on a variety of tumor-infiltrating immune cells, especially myeloid derived suppressor cells (MDSCs) and CD8⁺ T cells. Then, peptide C1 with binding affinity to VISTA was developed by phage displayed bio-panning technique, and its mutant peptide VS3 was obtained by molecular docking based mutation. Peptide VS3 could bind VISTA with high affinity and block its interaction with ligand PSGL-1 under acidic condition, and elicit anti-tumor activity in vivo. The peptide DVS3-Pal was further designed by d-amino acid substitution and fatty acid modification, which exhibited strong proteolytic stability and significant anti-tumor activity through enhancing CD8⁺ T cell function and decreasing MDSCs infiltration. This is the first study to develop peptides to block VISTA/PSGL-1 interaction, which could act as promising candidates for cancer immunotherapy.