Superantigen-fused T cell engagers for tumor antigen-mediated robust T cell activation and tumor cell killing.

Inadequate T cell activation has severely limited the success of T cell engager (TCE) therapy, especially in solid tumors. Enhancing T cell activity while maintaining the tumor specificity of TCEs is the key to improving their clinical efficacy. However, currently, there needs to be more effective strategies in clinical practice. Here, we design novel superantigen-fused TCEs that display robust tumor antigen-mediated T cell activation effects. These innovative drugs are not only armed with the powerful T cell activation ability of superantigens but also retain the dependence of TCEs on tumor antigens, realizing the ingenious combination of the advantages of two existing drugs. Superantigen-fused TCEs have been preliminarily proven to have good (>30-fold more potent) and specific (>25-fold more potent) antitumor activity in vitro and in vivo. Surprisingly, they can also induce the activation of T cell chemotaxis signals, which may promote T cell infiltration and further provide an additional guarantee for improving TCE efficacy in solid tumors. Overall, this proof-of-concept provides a potential strategy for improving the clinical efficacy of TCEs.

Improving the tumor selectivity of T cell engagers by logic-gated dual tumor-targeting.

Targeting single tumor antigens makes it difficult to provide sufficient tumor selectivity for T cell engagers (TCEs), leading to undesirable toxicity and even treatment failure, which is particularly serious in solid tumors. Here, we designed novel trispecific TCEs (TriTCEs) to improve the tumor selectivity of TCEs by logic-gated dual tumor-targeting. TriTCE can effectively redirect and activate T cells to kill tumor cells (∼18 pM EC50) by inducing the aggregation of dual tumor antigens, which was ∼70- or 750- fold more effective than the single tumor-targeted isotype controls, respectively. Further in vivo experiments indicated that TriTCE has the ability to accumulate in tumor tissue and can induce circulating T cells to infiltrate into tumor sites. Hence, TriTCE showed a stronger tumor growth inhibition ability and significantly prolonged the survival time of the mice. Finally, we revealed that this concept of logic-gated dual tumor-targeted TriTCE can be applied to target different tumor antigens. Cumulatively, we reported novel dual tumor-targeted TriTCEs that can mediate a robust T cell response by simultaneous recognition of dual tumor antigens at the same cell surface. TriTCEs allow better selective T cell activity on tumor cells, resulting in safer TCE treatment.

Hand Hygiene among Anesthesiologists and Microorganisms Contamination in Anesthesia Environments: A Single-Center Observational Study.

Objective: To investigate the baseline levels of microorganisms' growth on the hands of anesthesiologists and in the anesthesia environment at a cancer hospital. Methods: This study performed in nine operating rooms and among 25 anesthesiologists at a cancer hospital. Sampling of the hands of anesthesiologists and the anesthesia environment was performed at a ready-to-use operating room before patient contact began and after decontamination. Results: Microorganisms' growth results showed that 20% (5/25) of anesthesiologists' hands carried microorganisms (> 10 CFU/cm 2) before patient contact began. Female anesthesiologists performed hand hygiene better than did their male counterparts, with fewer CFUs ( P = 0.0069) and fewer species ( P = 0.0202). Our study also found that 55.6% (5/9) of ready-to-use operating rooms carried microorganisms (> 5 CFU/cm 2). Microorganisms regrowth began quickly (1 hour) after disinfection, and increased gradually over time, reaching the threshold at 4 hours after disinfection. Staphylococcus aureus was isolated from the hands of 20% (5/25) of anesthesiologists and 33.3% (3/9) of operating rooms. Conclusion: Our study indicates that male anesthesiologists need to pay more attention to the standard operating procedures and effect evaluation of hand hygiene, daily cleaning rate of the operating room may be insufficient, and we would suggest that there should be a repeat cleaning every four hours.

Practices, Attitudes, and Knowledge Among Healthcare Providers and Oncologists in China Regarding Male Fertility Preservation.

Objective: The purpose of this study was to help to promote a better understanding of the male fertility preservation status in China. Methods: In this cross-sectional survey, 1,912 healthcare providers and oncologists were surveyed anonymously using 16 questions carried out at community oncology practices in China from September 2018 to April 2021. 16 questions were designed to evaluate their knowledge on male fertility preservation in cancer patients, assess the factors they considered when deciding whether to discuss male fertility preservation with their patients. Results: Among the 1,912 healthcare providers (42.2% male), 1,713 (89.6%) considered that patients with cancer should be recommended for fertility preservation. 1,264 (66.1%) respondents were aware of male fertility preservation, but only 248 (13.0%) respondents knew the correct institutions. Whether a healthcare provide recommended fertility preservation to their patients depended on the provider's educational background, professional qualifications, hospital grade, area, department, and age. Among the healthcare providers, the three main factors for not recommending fertility preservation for patients with cancer were lack of suitability of the patient for fertility (28.2%), lack of knowledge of fertility preservation (28.6%), and lack of knowledge concerning the institutes that provide fertility preservation (25.4%). Conclusion: Despite this, healthcare providers and oncologists in China showed a positive attitude toward fertility preservation in patients with cancer. Hence, the education of physicians should include fertility preservation, with the aim of increasing their knowledge and awareness. There should be more collaboration between oncologists and reproductive medicine specialists.

Correlation of Moraxella catarrhalis macrolide susceptibility with the ability to adhere and invade human respiratory epithelial cells.

Recently, the prevalence of macrolide-resistant Moraxella catarrhalis has been reported, especially among Chinese children. The fitness cost of resistance is reported to render the resistant bacteria less virulent. To investigate the correlation between macrolide susceptibility of M. catarrhalis and pathogenicity, the whole genome of 70 M. catarrhalis isolates belonging to four clonal complexes with different macrolide susceptibilities was sequenced. The gene products were annotated with the Gene Ontology terms. Based on 46 extracted essential virulence genes, 19 representative isolates were selected to infect type II alveolar cells (A549 cells). The ability of these isolates to adhere and invade human epithelial cells and to produce cytokines was comparatively analysed. Furthermore, mice were infected with a pair of M. catarrhalis isolates with different pathogenic behaviours and macrolide susceptibilities to examine pulmonary clearance, histological findings, and the production of cytokines. The percentages of annotations for binding, metabolic process, cellular process, and cell were non-significantly different between the macrolide-resistant and macrolide-susceptible groups. The presence of uspA2, uspA2H, pilO, lbpB, lex1, modM, mboIA, and mboIB significantly differed among the four clonal complexes and macrolide susceptibility groups. Furthermore, compared with those in macrolide-susceptible isolates, the adhesion ability was stronger (P = 0.0019) and the invasion ability was weaker (P < 0.0001) in the macrolide-resistant isolates. Mouse experiments revealed that pulmonary macrophages elicit immune responses against M. catarrhalis infection by significantly upregulating the Csf2, Il4, Il13, Il1b, Il6, Tnf, and Il18. Therefore, M. catarrhalis populations exhibited diverse pathogenicity in vitro and in vivo.

Evaluation of Anti-Helicobacter pylori IgG Antibodies for the Detection of Helicobacter pylori Infection in Different Populations.

Serological testing (immunoassay) for Helicobacter pylori (H. pylori) is widely available and inexpensive, and does not require medication modifications before testing. It can also determine the type of infection, which helps with clinical diagnosis and treatment, and guides the use of medication. However, the performance of immunoblotting for the detection of H. pylori infections in different populations has still not been fully evaluated. We performed a retrospective analysis of patients in the Health Examination Center and Outpatient Department, from November 2017 to September 2020, at Peking Union Medical College Hospital. All the subjects were tested with the 13C-urea breath test (13C-UBT) and for IgG antibodies. A total of 1678 participants, including 1377 individuals who had undergone physical examinations, were recruited. The results of the immunoassay were significantly different from those of the 13C-UBT for all the subjects and outpatients (p < 0.001). For the physical examinations of individuals, the agreement between the immunoassay and the 13C-UBT was 0.64 (95%CI: 0.59−0.68; p < 0.001), and the H. pylori immunoassay demonstrated a sensitivity and specificity of 74.24% and 90.45%, respectively, with a positive predictive value of 71.01% and negative predictive value of 91.76%. In addition, in patients with gastric mucosal atrophy or early gastric cancer, antibody typing tests can also detect infected patients with missed UBT. The prevalence of H. pylori in Beijing was 26.8%, and the serological positivity rate for H. pylori in the population of Beijing was about 31.7% (25.1% in the physical examination population). The rate of H. pylori antibody positivity among patients with allergic diseases was 73.5%, which is significantly higher than that of the non-allergic disease population (29.3%, p < 0.001). In conclusion, H. pylori antibody typing testing can be applied as a specific test in the healthy physical examination population, and the test can be performed with the remaining serum during the physical examination.

Soluble Expression of Fc-Fused T Cell Receptors Allows Yielding Novel Bispecific T Cell Engagers.

The specific recognition of T cell receptors (TCR) and peptides presented by human leukocyte antigens (pHLAs) is the core step for T cell triggering to execute anti-tumor activity. However, TCR assembly and soluble expression are challenging, which precludes the broad use of TCR in tumor therapy. Herein, we used heterodimeric Fc to assist in the correct assembly of TCRs to achieve the stable and soluble expression of several TCRs in mammalian cells, and the soluble TCRs enable us to yield novel bispecific T cell engagers (TCR/aCD3) through pairing them with an anti-CD3 antibody. The NY-ESO-1/LAGE-1 targeted TCR/aCD3 (NY-TCR/aCD3) that we generated can redirect naïve T cells to specific lysis antigen-positive tumor cells, but the potency of the NY-TCR/aCD3 was disappointing. Furthermore, we found that the activation of T cells by NY-TCR/aCD3 was mild and unabiding, and the activity of NY-TCR/aCD3 could be significantly improved when we replaced naïve T cells with pre-activated T cells. Therefore, we employed the robust T cell activation ability of staphylococcal enterotoxin C2 (SEC2) to optimize the activity of NY-TCR/aCD3. Moreover, we found that the secretions of SEC2-activated T cells can promote HLA-I expression and thus increase target levels, which may further contribute to improving the activity of NY-TCR/aCD3. Our study described novel strategies for soluble TCR expression, and the optimization of the generation and potency of TCR/aCD3 provided a representative for us to fully exploit TCRs for the precision targeting of cancers.

Laboratory diagnosis of COVID-19 in China: A review of challenging cases and analysis.

Since the initial emergence of coronavirus disease 2019 (COVID-19) in Wuhan, Hubei province, China, a rapid spread of the disease occurred around the world, rising to become an international global health concern at pandemic level. In the face of this medical challenge threatening humans, the development of rapid and accurate methods for early screening and diagnosis of COVID-19 became crucial to containing the emerging public health threat, and prevent further spread within the population. Despite the large number of COVID-19 confirmed cases in China, some problematic cases with inconsistent laboratory testing results, were reported. Specifically, a high false-negative rate of 41% on severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) detection by real-time reverse transcription-polymerase chain reaction (qRT-PCR) assays was observed in China. Although serological testing has been applied worldwide as a complementary method to help identify SARS-CoV-2, several limitations on its use have been reported in China. Therefore, the use of both qRT-PCR and serological testing in the diagnosis of COVID-19 in China and elsewhere, presented considerable challenges, but when used in combination, can be valuable tools in the fight against COVID-19. In this review, we give an overview of the advantages and disadvantages of different molecular techniques for SARS-CoV-2 detection that are currently used in several labs, including qRT-PCR, gene sequencing, loop-mediated isothermal amplification (LAMP), nucleic acid mass spectrometry (MS), and gene editing technique based on clustered regularly interspaced short palindromic repeats (CRISPR/Cas13) system. Then we mainly review and analyze some causes of false-negative qRT-PCR results, and how to resolve some of the diagnostic dilemma.

Specific Inhibition of Tumor Growth by T Cell Receptor-Drug Conjugates Targeting Intracellular Cancer-Testis Antigen NY-ESO-1/LAGE-1.

Despite the significant therapeutic advances in T-cell immunotherapy, many malignancies remain unresponsive, which might be because of the negative regulation of T cells by the tumor microenvironment (TME). T cells discriminate tumor cells and normal cells through T-cell receptors (TCRs); therefore, we generated a novel type of TCR-drug conjugates (TDCs) by referring antibody-drug conjugations (ADCs) to overcome the effects of the TME on T cells while preserving the specificity of TCR for tumor recognition. We selected HLA-A2/NY-ESO-1157-165 (peptide NY-ESO-1157-165 in complex with human leukocyte antigen serotype HLA-A*02:01) as the antigen and the antigen-specific TCR (1G4113) as the carrier. By sortase A-mediated ligation, we obtained three NY-TCR-vcMMAEs and further studied their properties, antitumor activity, and toxicity in vitro and in vivo. We found that all the NY-TCR-vcMMAEs had high endocytosis efficiency and specifically killed HLA-A2/NY-ESO-1157-165 positive tumor cells. In xenograft models, one of the TDCs, NY-TCR-2M, was effectively and specifically distributed into tumor tissues and inhibited tumor growth without inducing obvious toxicity. Our results demonstrated that TCRs can be carriers of toxic payloads and that the TDCs thus formed can specifically inhibit tumor growth, neglecting the immune microenvironment.

The Antitumor Activity of TCR-Mimic Antibody-Drug Conjugates (TCRm-ADCs) Targeting the Intracellular Wilms Tumor 1 (WT1) Oncoprotein.

Wilms tumor 1 (WT1) oncoprotein is an intracellular oncogenic transcription factor which is barely expressed in normal adult tissues but over expressed in a variety of leukemias and solid cancers. WT1-derived HLA-A*02:01 T cell epitope, RMFPNAPYL (RMF), is a validated target for T cell-based immunotherapy. We generated two T cell receptor mimic antibody-drug conjugates (TCRm-ADCs), ESK-MMAE, and Q2L-MMAE, against WT1 RMF/HLA-A*02:01 complex with distinct affinities, which mediate specific antitumor activity. Although ESK-MMAE showed higher tumor growth inhibition ratio in vivo, the efficacy of them was not so promising, which might be due to low expression of peptide/HLA targets. Therefore, we explored a bispecific TCRm-ADC that exerted more potent tumor cytotoxicity compared with TCRm-ADCs. Hence, our findings validate the feasibility of the presenting intracellular peptides as the targets of ADCs, which broadens the antigen selection range of antibody-based drugs and provides new strategies for precision medicine in tumor therapy.

Effects of ethylene and NO on AsA-GSH in lotus under cadmium stress.

Under the background of Cd (50 µmol·L-1) stress, we added ethylene precursor ACC (100 µmol·L-1), ACC + nitric oxide synthase (NOS) inhibitor L-NNA (200 µmol·L-1), ACC + nitrate reductase (NR) inhibitor Tu (1 mmol·L-1), ACC + nitric oxide (NO) scavenger PTIO (200 µmol·L-1), NO donor SNP (500 µmol·L-1), SNP + ethylene signal inhibitor STS (100 µmol·L-1) to examine their effects on the damage degree of leaves and response mechanisms of AsA-GSH cycle in lotus 'Weishanhuhonglian'. Results showed toxic symptom of lotus leaves under Cd stress. The relative conductivity, malondialdehyde (MDA), as well as ascorbic acid (AsA) and glutathione (GSH) contents were significantly increased, but the activities of ascorbate peroxidase (APX), glutathione reductase (GR), monodehydroascorbate reductase (MDHAR) and dehydroascorbate reductase (DHAR) were obviously decreased. Compared with Cd stress, adding ACC significantly increased the damage area of lotus leaves, decreased activities of the above-mentioned four antioxidant enzymes and increased AsA and GSH contents. SNP aggravated the toxic symptom of lotus leaves and decreased GR and MDHAR activities. PTIO significantly relieved the toxic symptom of leaves, increased activities of APX, GR, MDHAR and DHAR, but decreased AsA and GSH contents compared with Cd and ACC treatment. However, the effects of L-NNA and Tu were not as obvious as PTIO's. In comparison with Cd and SNP treatment, STS relieved the toxic symptom of leaves, increased APX, GR, MDHAR and DHAR activities, and decreased AsA and GSH contents. Taken together, these results showed the synergistic effects of ethylene and NO in regulating lotus responses to Cd stress through AsA-GSH cycle.

18F-FDG PET/CT for the early prediction of the response rate and survival of patients with recurrent or metastatic breast cancer.

The present study aimed to explore the value of fludeoxyglucose F 18 positron emission tomography-computed tomography (PET/CT) for the early prediction of chemotherapy remission rates and survival in patients with recurrent and metastatic breast cancer. A total of 24 patients diagnosed with recurrent or metastatic breast cancer between 2009 and 2014 were enrolled. All patients underwent a PET/CT examination prior to (PET/CT1) and following (PET/CT2) chemotherapy. Differences of PET/CT1 maximal standardized uptake values (SUVmax), PET/CT2 SUVmax, ΔSUVmax and the ΔSUVmax% between objective remission (OR) and non-OR groups were measured. Survival differences between OR and non-OR groups and the overall survival (OS) between metabolic responsive and metabolic non-responsive groups were analyzed. In the present study, it was revealed that ΔSUVmax and ΔSUVmax% were significantly higher in the OR group compared with the non-OR group (P<0.001). Overall survival was significantly prolonged in the OR and metabolic responder groups compared with their respective control groups (P<0.001 and P<0.01, respectively). ΔSUVmax% were significantly positively associated with OS (r2=0.266; P<0.01). In conclusion, PET/CT may be valuable for the early prediction of the chemotherapy efficacy and survival of patients with recurrent or metastatic breast cancer.

Elimination of melanoma by sortase A-generated TCR-like antibody-drug conjugates (TL-ADCs) targeting intracellular melanoma antigen MART-1.

Most tumor-associated proteins are located inside tumor cells and thus are not accessible to current marketed therapeutic monoclonal antibodies or their cytotoxic conjugates. Human leukocyte antigen (HLA) class I can present peptides derived from intracellular tumor-associated proteins and somatically mutated proteins on the cell's surface, forming an HLA/peptide complex as tumor-specific antigens for T cell receptor (TCR) recognition. Therefore, HLA-mediated presentation of intracellular tumor antigen peptides provides a viable way to distinguish tumor cells from normal cells, which is important for broadening antigen selection, especially for antibody-drug conjugates (ADCs) regarding their highly cytotoxic payload. We applied sortase A-mediated conjugation to develop TCR-like ADCs (i.e., EA1 HL-vcMMAE) targeting intracellular MART-1 protein, a melanocyte-differentiating antigen specific for metastatic melanomas, via the cell surface HLA-A2/MART-126-35 peptide complex. Homogenous EA1 HL-vcMMAE (drug to antibody ratio of 4) efficiently eliminated melanoma cells in xenograft mouse models with no obvious toxicity at the therapeutic dosage. Trametinib, an MEK inhibitor serving as an HLA expression enhancing agent, augmented the TL-ADCs' efficacy both in vitro and in vivo by upregulating MART-126-35 peptide presentation, thus providing a strategy for overcoming the limitation of antigen presentation level for TL-ADCs. Hence, our findings validate the strategy of using sortase A-generated TL-ADCs to target tumor-specific intracellular proteins, with or without agents present, to increase presenting TCR epitope peptides.

In situ quantitative bioanalysis of monomethyl auristatin E-conjugated antibody-drug conjugates by flow cytometry.

Antibody-drug conjugates (ADCs) consist of cytotoxic agents covalently conjugated to monoclonal antibodies that substantially improve antitumour activity and reduce systemic toxicity. With the growing number of ADCs in clinical applications, more accurate bioanalysis data are urgently needed to facilitate the development and rational use of ADCs. Herein, we used antigen-positive cells as antigen carriers and ofatumumab (OFA-HL) and ofatumumab-based ADC (OFA-HL-MMAE) as examples to establish a new ligand-binding assay (LBA) method based on flow cytometry. We proved that the new method met the required analytical performance criteria and the lower limit of quantitation (LOQ) was 0.2 µg/mL. In addition, the LOQ of the quantitative OFA-HL flow cytometry method was reduced to 0.025 µg/mL by choosing an optimized fluorescent antibody, which indicated that the LOQ of the new method can be improved. What's more, the new method showed good stability and specificity when we used it to determine the concentrations of OFA-HL and OFA-HL-MMAE in mouse serum. During the bioanalysis of ADCs, various factors should be considered. Therefore, choosing optimal methods for ADC bioanalysis is necessary. This new method using in situ antigens not only extends the scope of application of the conventional LBA methods by avoiding the need for soluble antigens, but also improves the authenticity of ADC bioanalysis as a supplementary approach, which is valuable for developing accurate ADC assays.

Profiling of PDR1 and MSH2 in Candida glabrata Bloodstream Isolates from a Multicenter Study in China.

Among 158 Candida glabrata bloodstream isolates collected from numerous centers in China, a resistance to fluconazole was seen in 8.9%. Three isolates (1.9%) were resistant to all echinocandins. Multilocus sequence typing (MLST) revealed that sequence type 7 ([ST7] 65.8%) was the most common type, followed by ST3 (7.6%). PDR1 polymorphisms were associated with the acquisition of fluconazole resistance in C. glabrata isolates, while MSH2 polymorphisms were associated with the STs and microsatellite genotypes, irrespective of fluconazole resistance.

Clinical characteristics of the first cases of invasive candidiasis in China due to pan-echinocandin-resistant Candida tropicalis and Candida glabrata isolates with delineation of their resistance mechanisms.

Echinocandin antifungal agents have become the first-line therapy for invasive candidiasis (IC) in many countries. Despite their increasing use, resistance to this class of drug is, overall, still uncommon. Here, we report two patients from the People's Republic of China with IC, one with infection caused by pan-echinocandin-resistant Candida tropicalis and the other by pan-echinocandin-resistant Candida glabrata. We also describe the mechanisms of drug resistance of these isolates. The echinocandin-resistant C. glabrata isolate was cultured from ascitic fluid of a 46-year-old male patient with intra-abdominal IC developing after surgery in 2012. This patient had had no prior antifungal exposure. The echinocandin-resistant C. tropicalis isolate was cultured from chest drainage fluid of a 60-year-old female patient with severe coronary disease and lung infection. Prior to culture and identification of the isolate, the patient had received micafungin treatment for 19 days. Both isolates were cross-resistant to micafungin, anidulafungin, and caspofungin, with minimum inhibitory concentration values of ≥2 µg/mL. The amino acid substitution E655K was found adjacent to the FKS2 HS1 region of the C. glabrata isolate, while the substitution S80P were found in the FKS1 HS1 region of the C. tropicalis isolate. This report highlights the emergence of echinocandin resistance in two important non-albicans Candida species. Although the overall prevalence of echinocandin resistance is low in the People's Republic of China, monitoring of antifungal susceptibility trends in all Candida species is warranted.

[Infection Status of Human Papilloma Virus,Ureaplasma Urealyticum, Chlamydia Trachomatis,and Neisseria Gonorrhoeae].

Objective To analyze the infection status of human papilloma virus (HPV),Ureaplasma urealyticum (UU),Chlamydia trachomatis (CT),and Neisseria gonorrhoeae (NG) in clinical patients.Methods The laboratory specimens including urine,urethral swabs,and cervical swabs from 870 patients from January 1st 2014 to December 31st 2017 were retrospectively analyzed. HPV-DNA was detected by multiplex fluorescent PCR,and the UU-RNA,CT-RNA,and NG-RNA were determined by isothermal nucleic acid amplification. The positive rate of each pathogen and the distribution of positive rate between male and female patients were calculated. The samples were further divided into HPV-positive group and HPV-negative group,and the positive rates of UU-RNA,CT-RNA,and NG-RNA in these two groups were compared.Results The highest positive rate was 53.68%(467/870) for UU-RNA,followed by HPV-DNA [32.41%(282/870) ]and NG-RNA [2.18%(19/870)]. The total positive rate of high-risk (HR)-HPV(subtypes:16,18,31,33,35,39,45,51,52,56,58,59,and 68) [31.52%(209/663)]and UU in female patients [60.93%(404/663)] was significantly higher than that in male patients [17.39%(36/207),30.34%(63/207)](both P<0.001). The male patients had significantly higher CT positive rate in HR-HPV-positive group than in HR-HPV-negative group [22.58%(7/31) vs. 4.54%(8/176)](P<0.001). The female patients had significantly higher CT positive rate in HR-HPV-positive group than in HR-HPV-negative group [10.5%(21/200) vs. 5.61%(26/463)](P=0.024). The UU-RNA positive rate of females in the low-risk (LR)-HPV (subtypes:6 and 11) positive group was significantly higher than that in LR-HPV negative group [70.83%(34/48) vs.2.11%(13/615)](P<0.001).Conclusions Women are more susceptible to HR-HPV and UU infections. HR-HPV-positive patients are more likely to experience CT infection. In contrast,co-infection with UU is more common in LR-HPV-positive females.

Tumor tissue katanin P60 expression correlates with lymph node metastasis and worse prognosis in patients with breast cancer: A cohort study.

OBJECTIVE: To evaluate the correlation of katanin P60 expression with clinicopathological grade and overall survival (OS) in patients with breast cancer (BC). METHODS: Three hundred and four operative BC patients were retrospectively reviewed in this cohort study. Tumor tissue sample was acquired from each patient during surgery. Immunofluorescent staining was used to assess katanin P60 expression. RESULTS: There were 265 BC patients with katanin P60 negative expression and 75 patients with katanin P60 positive expression. Higher N stage (p< 0.001) and TNM stage (p< 0.001) were observed in katanin P60 positive expression group compared to katanin P60 negative expression group. Kaplan-Meier (K-M) curves showed association of katanin P60 positive expression status with shorter OS. Multivariate Cox analysis illustrated katanin P60 positive expression (p< 0.001) could independently predict worse OS. Subgroups analysis indicated that katanin P60 positive expression correlated with worse OS in patients of TNM stage II (p= 0.001) and stage III (p=0.001), while no correlation was found between katanin P60 expression and OS in BC patients with stage I (p= 0.538). According to molecular subtypes, katanin P60 positive expression was correlated with shorter OS in patients with HER2+HR+ (p< 0.001), HER2+HR- (p< 0.001) and HER2-HR- (p= 0.001) status compared to katanin P60 negative expression, while no correlation between katanin P60 expression and OS was observed in patients with HER2-HR+ (p= 0.073). CONCLUSION: Katanin P60 positive expression was correlated with higher lymph node metastasis and worse OS, and it could be regarded as a novel and convincing biomarker to independently predict the prognosis of BC patients.

[Effects of Traffic-related Air Pollution Exposure on DNA Methylation].

The goal of the present study was to explore the effects of traffic-related air pollution exposure on DNA methylation. Into five groups of 6, 30 healthy Wistar rats were randomly divided. Three groups of rats were then exposed to traffic-related air pollution at high (tunnel), moderate (crossroad), and low (control) pollution levels for 7 d, whereas the two other groups were exposed in the tunnel for 14 d/28 d. The levels of PM10 and NO2 were measured during the exposure. The study was performed in spring and autumn, and lung tissue and blood were collected after the exposure. Promoter methylation levels of p 53 , MGMT, and MAGE-A 4 were quantified via pyrosequencing. The levels of PM10 and NO2 in the crossroad and tunnel groups were significantly higher than those in the control group. After 7 d exposure in autumn, promoter methylation levels of p 53 and MGMT in lung tissue significantly decreased, and the methylation status continued to decrease with increasing exposure time; MAGE-A 4 was highly methylated and showed no difference among the three groups. DNA methylation in lung tissue was more likely to be changed compared with that in blood during 7 d exposure. As the exposure time increased, DNA methylation changes between blood and lung tissue started to coincide. In lung tissue, PM10 exposure was significantly associated with decreased p 53 promoter methylation (r=-0.347, P=0.038) and NO2 exposure was significantly associated with decreased promoter methylation of p 53, MGMT, and MAGE-A 4 (r=-0.482, -0.444, and -0.346, respectively; P< 0.05). In blood, PM10 and NO2 were significantly and positively associated with MAGE-A 4 promoter methylation (r=0.395 and 0.431, respectively; P< 0.05). Traffic-related air pollution exposure may induce promoter hypomethylation of p 53 and MGMT.