OBJECTIVE To discuss the gentamycin-induced ototoxity and mechanism on cochlear outer hair cells. METHODS All the guinea pigs were randomly divided into 3 groups. Gentamycin, gentimycin plus furosemide and normal saline were intraperitoneally injected for 6 days respectively. The amplititude of distortion product otoacoustic emissions (DPOAE)urface prepration of cochlear basilar membrane, hair cells’ apoptosis and expression of caspase-3 after injection were analyzed. RESULTS Reduction of the DPOAE amplitude, hair cell damage, apoptosis and Caspase-3 expression were observed in group injected with gentamycin as well as in group injected with gentamycin plus furosemide. CONCLUSION Gentamycin should induce hair cell damage in the cochlea of guinea pigs and Caspase-3 plays an important role in the process of hair cell apoptosis.

Fluorogenic quantitative polymerase chain reaction(FQ-PCR) is an emerging technique with high specificity,susceptibility,automatism and has been widely used for the detection and quantification of microorganism,some genetic diseases and cancer.The design and establishment of standard FQ-PCR laboratory are required not only by gene expansion and diagnosis but also by biosafty and veracity.The aim of the paper is to introduce the structural characteristics,instruments requirement and comfort parameters of FQ-PCR laboratory.

Abstract Gram-negative ( G-) bacteria, such as denitrifying bacteria and anaerobic ammonia oxidation bacteria, are highly social organisms capable of sophisticated cooperative behavior mediated via quorum sensing. As signal molecules of the chemical communication, N-acyl-homoserine lactones ( AHLs ) can mediate the quorum sensing of the functional microbial population and regulate the population density. To understand the growth of functional microbial population and the mechanism for biological nitrogen removal in upflow microaerobic sludge reactors ( UMSRs ) treating organic wastewater with low ratio of chemical oxygen demand to total nitrogen, a method was established to simultaneously detect AHLs in the microaerobic processes. Water-sludge mixtures sampled from the UMSRs were pretreated in sequence by liquid-liquid extraction using ethyl acetate, rotary evaporation, constant volume with methanol, separation by C18 column. Gradient elution was carried out using 5 mmol/L ammonium acetate ( containing 0 . 1% formic acid ) and methanol as mobile phases. On the base of multiple reaction monitoring analysis, a triple quadrupole mass spectrometer with an electrospray ionization was introduced to detect the target compounds. Nine kinds of AHLs were used to evaluate the established method and the results showed that the detection limits were 0 . 01-0 . 5 μg/L and all of the AHLs presented excellent linearity with the concentration ranging from 0 . 5 to 100 μg/L. The recovery and relative standard deviation ranged from 62. 5% to 118. 1% and 2. 9% to 12. 1%, respectively. The analysis could be finished within 6. 5 min. The rapid, accurate and precise method for detecting AHLs provided a new insight into the growth and metabolic activity of functional microbial population in the activated sludge processes to understand the mechanism of biological nitrogen removal, suggesting a good application in regulation and operation of wastewater biological treatment processes.

AIM:To establish and characterize the patient-derived esophageal squamous-cell carcinoma xeno-graft (PDECX) in mice.METHODS:The samples of human esophageal cancer were grafted into severe combined immu-nodeficient ( SCID) mice.The xenografts were transferred to SCID mice when the first passage of xenografts grew up .The growth of tumors in the first, second and third passages was observed .HE staining was performed.The expression of CK5/6, p63 and p40 in the patient samples , and the first and third passages of the xenografts were detected by immunohisto-chemical analysis.The expression of mTOR, p-mTOR, p70S6K, p-p70S6K, Akt1, p-Akt (Ser473), Erk1/2 and p-Erk1/2 were determined by Western blot .RESULTS:The PDECX was successfully established .The positive expression of CK5/6, p63 and p40 in the xenografts was consistent with that in the patients ’ samples.The levels of phosphorylated and total proteins of proliferation-related signaling pathways were different in the xenografts from different patients .CONCLU-SION:The PDECX model adequately reflects the tumal heterogeneity that is observed in the patients .

Objective: To investigate the fatigue status of military personnel stationed in plateau and high cold region, and to analyze the mediator effect of trait coping style on job stress and fatigue. Methods: In October 2010, with the method of cluster random sampling survey, 531 military personnel stationed in plateau and high cold region were chosen as subject. The fatigue status were evaluated by the Chinese version multidimensional fatigue inventory (MFI-20) , job stress were evaluated by the Job Stress Survey (JSS) , and trait coping style were evaluated by the Trait Coping Style Questionnaire (TCSQ) . Results: According to the information of different population characteristics, mean rank of physical fatigue about the urban (town) group were higher than that of rural group (Z=-2.200, P<0.05) ; mean rank of reduced motivation about the urban (town) group were higher than that of rural group (Z=-2.781, P<0.05) ; mean rank of general fatigue scores about the urban (town) group were higher than that of rural group (Z=-3.026, P<0.05) ; mean rank of physical fatigue about the up or equal 20-years old age group were higher than that of below 20-years old age group (Z=-4.045, P<0.05) ; mean rank of reduced motivation about the up or equal 20-years old age group were higher than that of below 20-years old age group (Z=-2.182, P<0.05) ; mean rank of mental fatigue about the up or equal 20-years old age group were higher than that of below 20-years old age group (Z=-2.879, P<0.05) ; mean rank of general fatigue scores about the up or equal 20-years old age group were higher than that of below 20-years old age group (Z=-3.647, P<0.05) ; mean rank of reduced motivation were significant statistical difference among the military officers, sergeancy and soldier group (F=18.965, P<0.05) ; mean rank of general fatigue scores were significant statistical difference among the military officers, sergeancy and soldier group (F=14.711, P<0.05) . The score of negative coping style were positively correlated with the score of physical fatigue (r_s=0.129) , reduced activity (r_s=0.123) , reduced motivation (r_s=0.149) and general fatigue (r_s=0.174) respectively, the score of organizational support lack strength were positively correlated with the score of physical fatigue (r_s=0.090) , reduced activity (r_s=0.098) , reduced motivation (r_s=0.099) and general fatigue (r_s=0.130) respectively. The mediator effect of negative coping style on the job stress and fatigue was 0.013 (P<0.01) . Conclusion The fatigue statuses of the urban (town) group and the up or equal 20-years old age group are poor, and the negative coping style plays mediator effect on the job stress and fatigue.

Drugs targeting immune checkpoint are used for cancer treatment, but resistance to single drug may occur. Combination therapy blocking multiple checkpoints simultaneously can improve clinical outcome. Therefore, we designed a recombinant protein rPC to block multiple targets, which consists of extracellular domains of programmed cell death protein 1 (PD-1) and cytotoxic T lymphocyte-associated antigen 4 (CTLA-4). The coding sequence was inserted into expression vector and stably transfected into HEK293 cells. The culture supernatant was collected and rPC was affinity-purified. Real-time quantitative PCR was used to evaluate the expression levels of ligands for PD-1 and CTLA-4 in several human cancer cell lines. The binding of rPC with cancer cells was examined by immunofluorescence cell staining, the influence of rPC on cancer cell growth was assayed by CCK-8. The results showed that rPC could be expressed and secreted by stably transfected HEK293 cells, the purified rPC could bind to lung cancer NCI-H226 cells which have high levels of ligands for PD-1 and CTLA-4, no direct impact on cancer cell growth could be observed by rPC treatment. The recombinant protein rPC can be functionally assayed further for developing novel immunotherapeutic drugs for cancer.
Animals ; CTLA-4 Antigen ; genetics ; Cell Proliferation ; HEK293 Cells ; Humans ; Lung Neoplasms ; metabolism ; Programmed Cell Death 1 Receptor ; genetics ; Protein Binding ; Protein Domains ; genetics ; Recombinant Fusion Proteins ; genetics ; isolation & purification ; metabolism

Objective: To study the effect of WT1 expression on the prognosis of allogeneic hematopoietic stem cell transplantation (allo-HSCT) in acute leukemia (AL) and its significance as molecular marker to dynamically monitor minimal residual disease (MRD) . Methods: Retrospectively analyzed those AL patients who underwent allo-HSCT in the First Hospital Affiliated to Zhejiang University School of Medicine during Jan 2016 to Dec 2017, a total number of 314 cases, 163 males and 151 females, median age was 30 (9-64) years old. Comparing the difference of WT1 expression at diagnosed, pre-HSCT and after HSCT. Using the receiver operating characteristic (ROC) curve to determine the WT1 threshold at different time so as to predict relapse. The threshold of WT1 expression before transplantation was 1.010%, within 3 months after HSCT was 0.079% and 6 months after HSCT was 0.375%. According to these thresholds, WT1 positive patients were divided into low expression groups and high expression groups. Analyzed the relationship between overall survival (OS) , disease-free survival (DFS) , cumulative incidence of relapse (CIR) and WT1 expression. Results: The OS and DFS of high expression group pre-HSCT were lower than low expression group [69.2% (9/13) vs 89.1% (57/64) , χ²=4.086, P=0.043; 53.8% (7/13) vs 87.5% (56/64) , χ²=9.766, P=0.002], CIR was higher than low expression group [30.8% (4/13) vs 7.8% (5/64) , P=0.017]. There was no significant difference of OS and DFS between high expression and low expression group of 3 months after HSCT (P=0.558, P=0.269) . The OS and DFS of high expression group of 6 months after transplantation were both lower than low expression group (P=0.049, P=0.035) . Multivariate analysis showed that WT1>0.375% when 6 months after transplantation was the only independent prognostic factor for shorter DFS (P=0.022) . There was no statistically significant difference in CIR between the high-expression group and the low-expression group 3 months after transplantation and 6 months after transplantation (P=0.114, P=0.306) . Conclusion High expression of WT1 before and after HSCT was an adverse prognosis factor. It is of clinical practical value to use WT1 as a transplant recommendation index for patients with acute leukemia and as a marker to monitor MRD dynamically.

OBJECTIVE To explore the mechanism of action of epigallocatechin-3-gallate(EGCG) in inhibiting laryngeal cancer cells. METHODS The expression of epidermal growth factor receptor(EGFR) in laryngeal cancer cell lines AMC-HN-8, TU686 and TU212 was detected by Western blotting, and the inhibitory effects of cetuximab and EGCG on three laryngeal cancer cells were detected by CCK-8 assay. A lentiviral vector containing EGFR promoter and Luc reporter gene was constructed to generate a TU686-EGFR-Luc cell line that could steadily express Luc activity. Luciferase assay was performed to evaluate the effect of EGCG on the transcription activity of EGFR promoter. Cell cycle and apoptosis of EGCG-treated laryngeal carcinoma cells were analyzed by flow cytometry, and changes of the levels of EGFR and downstream ERK1/2, cell cycle-associated proteins P53 and P27, apoptosis-associated proteins BCL2 and PART, and autophagy marker LC3A/B were further examined. RESULTS The laryngeal carcinoma cell lines were insensitive to cetuximab but could be effectively suppressed by EGCG. EGCG effectively inhibited the transcription activity of EGFR promoter. Treatment of TU686 cells at sub-IC50 dose EGCG resulted in significant cell cycle arrest at S phase with partial apoptosis. Significant inhibition of expression and activation of EGFR and downstream signaling pathway were observed. CONCLUSION EGCG can effectively downregulate EGFR and suppress laryngeal carcinoma cells, further investigation on in vivo effect and mechanisms are anticipated.

The stability of marginal bone level is an important factor in ensuring long-term good clinical efficacy of implantation.The design of implant-abutment interface(IAI)is of considerable importance,including whether undergoing multiple abutment disconnection,whether platform switching is used,connection type,connection torque,and connection material differences.These fac-tors can affect the micro gap and micro movement of the connection interface,which change the size of micro leakage and mechanical stability,thereby affecting the long-term clinical efficacy of implants.This article presents a detailed review of the progress of research on different designs in implant-abutment interface,in order to provide a theoretical basis for the selection and operation of abutments in clinical diagnosis and treatment.

Objective To investigate the risk factors for spontaneous bacterial peritonitis (SBP) in patients with cirrhotic ascites, and to establish a new model for predicting the development of SBP. Methods A total of 215 patients who were diagnosed with cirrhotic ascites in Hebei General Hospital from September 2016 to September 2020 were enrolled, and according to the presence or absence of SBP, they were divided into SBP group with 55 patients and non-SBP group with 160 patients. Related clinical data were collected and albumin-bilirubin (ALBI) score, Model for End-Stage Liver Disease (MELD) score, MELD combined with serum sodium concentration (MELD-Na) score, and Child-Pugh score were calculated. The t -test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups; a multivariate logistic regression analysis was used to screen out independent risk factors, and the receiver operating characteristic (ROC) curve was plotted to evaluate the performance of ALBI score, procalcitonin (PCT), polymorphonuclear neutrophil (PMN) count in ascites, and the ALBI-PMN-PCT combined model in the diagnosis of SBP. Results Compared with the SBP group, the non-SBP group had a significantly higher concentration of Na + ( Z =-3.414, P =0.001) and significantly lower total bilirubin ( Z =-2.720, P =0.007), creatinine ( Z =-1.994, P =0.046), urea nitrogen ( Z =-2.440, P =0.015), C-reactive protein ( Z =-9.137, P < 0.001), PCT ( Z =-8.096, P < 0.001), prothrombin time ( Z =-1.969, P =0.049), international normalized ratio ( Z =-2.073, P =0.038), PMN ( Z =-8.292, P < 0.001), MELD score ( Z =-2.736, P =0.006), MELD-Na score ( Z =-3.188, P =0.001), Child-Pugh score ( Z =-3.419, P =0.001), and ALBI score ( t =-5.010, P < 0.001), and there were also significant differences between the two groups in the presence or absence of gastrointestinal bleeding or hepatic encephalopathy ( χ 2 =16.551 and 8.142, P < 0.001 and P =0.004). The multivariate logistic regression analysis showed that ALBI score (odds ratio [ OR ]=3.460, 95% confidence interval [ CI ]: 1.296-9.240, P =0.013), PMN ( OR =1.012, 95% CI : 1.007-1.017, P < 0.001), and PCT ( OR =6.019, 95% CI : 2.821-12.843, P < 0.001) were independent risk factors for SBP in patients with cirrhotic ascites. The ROC curve showed that ALBI, PCT, PMN, and ALBI-PMN-PCT had areas under the ROC curve of 0.711, 0.866, 0.875, and 0.934, respectively, in the diagnosis of SBP, with sensitivities of 50.91%, 73.36%, 72.73%, and 89.09%, respectively, and specificities of 86.87%, 81.25%, 100.00%, and 91.87%, respectively. The patients with ALBI-PMN-PCT > 0.272 had an increased risk of developing SBP. Conclusion The ALBI-PMN-PCT combined model has a high value in predicting the onset of SBP in patients with cirrhotic ascites.