Adjuvant medial versus entire supraclavicular lymph node irradiation in high-risk early breast cancer (SUCLANODE): a protocol for a multicenter, randomized, open-label, phase 3 trial.

BACKGROUND: Supraclavicular nodal (SCL) irradiation is commonly used for patients with high-risk breast cancer after breast surgery. The Radiation Therapy Oncology Group (RTOG) and European Society for Radiotherapy and Oncology (ESTRO) breast contouring atlases delineate the medial part of the SCL region, while excluding the posterolateral part. However, recent studies have found that a substantial proportion of SCL failures are located in the posterolateral SCL region, outside of the RTOG/ESTRO-defined SCL target volumes. Consequently, many radiation oncologists advocate for enlarging the SCL irradiation target volume to include both the medial and posterolateral SCL regions. Nevertheless, it remains uncertain whether adding the posterolateral SCL irradiation improves survival outcomes for high-risk breast cancer patients. METHODS: The SUCLANODE trial is an open-label, multicenter, randomized, phase 3 trial comparing the efficacy and adverse events of medial SCL irradiation (M-SCLI group) and medial plus posterolateral SCL irradiation (entire SCL irradiation, E-SCLI group) in high-risk breast cancer patients who underwent breast conserving-surgery or mastectomy. Patients with pathological N2-3b disease following initial surgery, or clinical stage III or pathological N1-3b if receiving neoadjuvant systemic therapy, are eligible and randomly assigned (1:1) to M-SCLI group and E-SCLI group. Stratification is by chemotherapy sequence (neoadjuvant vs. adjuvant), T stage (T3-4 vs. T1-2), N stage (N1-2 vs. N3), and ER status (positive vs. negative). Other radiation volumes are identical in the two arms, including breast/chest wall, undissected axillary lymph node, and internal mammary node. Advanced intensity modulated radiation therapy (IMRT), volumetric modulated arc therapy (VMAT), or tomotherapy techniques are recommended. Both hypofractionated and conventional fractionation schedules are permitted. The primary end point is invasive disease-free survival, and secondary end points included overall survival, SCL recurrence, local-regional recurrence, distance recurrence, safety outcome, and patient-reported outcomes. The target sample size is 1650 participants. DISCUSSION: The results of the SUCLANODE trial will provide high-level evidence regarding whether adding posterolateral SCL irradiation to medial SCL target volume provides survival benefit in patients with high-risk breast cancer. TRIAL REGISTRATION: ClinicalTrials.gov Identifier: NCT05059379. Registered 28 September 2021, https://www. CLINICALTRIALS: gov/ct2/show/NCT05059379 .

RNA-sequencing-based detection of human viral pathogens in cerebrospinal fluid and serum samples from children with meningitis and encephalitis.

Encephalitis and meningitis are notable global public health concerns, especially among infants or children. Metagenomic next-generation sequencing (mNGS) has greatly advanced our understanding of the viruses responsible for these diseases. However, the detection rate of the aetiology remains low. We conducted RNA sequencing and virome analysis on cerebrospinal fluid (CSF) and serum samples commonly used in the clinical diagnosis to detect viral pathogens. In total, 226 paired CSF and serum samples from 113 children with encephalitis and meningitis were enrolled. The results showed that the diversity of viruses was higher in CSF, with a total of 12 viral taxa detected, including one case each of herpesvirus, coronavirus and enterovirus, and six cases of adenovirus related to human diseases. In contrast, the Anelloviridae was the most abundant viral family detected in serum, and only a few samples contained human viral pathogens, including one case of enterovirus and two cases of adenovirus. The detection rate for human viral pathogens increases to 10.6 %(12/113) when both types of samples are used simultaneously, compared to CSF along 7.9 % (9/113) or serum alone 2.6 % (3/113). However, we did not detect these viruses simultaneously in paired samples from the same case. These results suggest that CSF samples still have irreplaceable advantages for using mNGS to detect viruses in patients with meningitis and encephalitis, and serum can supplement to improve the detection rate of viral encephalitis and meningitis. The findings of this study could help improve the etiological diagnosis, clinical management and prognosis of patients with meningitis and encephalitis in children.

Etoposide, dexamethasone, and pegaspargase with sandwiched radiotherapy in early-stage natural killer/T-cell lymphoma: A randomized phase III study.

Methotrexate, etoposide, dexamethasone, and pegaspargase (MESA) with sandwiched radiotherapy is known to be effective for early-stage extranodal natural killer/T-cell lymphoma, nasal type (NKTCL). We explored the efficacy and safety of reduced-intensity, non-intravenous etoposide, dexamethasone, and pegaspargase (ESA) with sandwiched radiotherapy. This multicenter, randomized, phase III trial enrolled patients aged between 14 and 70 years with newly diagnosed early-stage nasal NKTCL from 27 centers in China. Patients were randomly assigned (1:1) to receive ESA (pegaspargase 2,500 IU/m2 intramuscularly on day 1, etoposide 200 mg orally, and dexamethasone 40 mg orally on days 2-4) or MESA (methotrexate 1 g/m2 intravenously on day 1, etoposide 200 mg orally, and dexamethasone 40 mg orally on days 2-4, and pegaspargase 2,500 IU/m2 intramuscularly on day 5) regimen (four cycles), combined with sandwiched radiotherapy. The primary endpoint was overall response rate (ORR). The non-inferiority margin was -10.0%. From March 16, 2016, to July 17, 2020, 256 patients underwent randomization, and 248 (ESA [n = 125] or MESA [n = 123]) made up the modified intention-to-treat population. The ORR was 88.8% (95% confidence interval [CI], 81.9-93.7) for ESA with sandwiched radiotherapy and 86.2% (95% CI, 78.8-91.7) for MESA with sandwiched radiotherapy, with an absolute rate difference of 2.6% (95% CI, -5.6-10.9), meeting the non-inferiority criteria. Per-protocol and sensitivity analysis supported this result. Adverse events of grade 3 or higher occurred in 42 (33.6%) patients in the ESA arm and 81 (65.9%) in the MESA arm. ESA with sandwiched radiotherapy is an effective, low toxicity, non-intravenous regimen with an outpatient design, and can be considered as a first-line treatment option in newly diagnosed early-stage nasal NKTCL.

Single-Cell RNA-Seq Analysis Reveals Ferroptosis in the Tumor Microenvironment of Clear Cell Renal Cell Carcinoma.

In this study, we investigated the role of ferroptosis in the tumor microenvironment (TME) of clear cell renal cell carcinoma (ccRCC), the leading cause of renal cancer-related death. We analyzed single-cell data from seven ccRCC cases to determine cell types most correlated with ferroptosis and performed pseudotime analysis on three myeloid subtypes. We identified 16 immune-related ferroptosis genes (IRFGs) by analyzing differentially expressed genes between cell subgroups and between high and low immune infiltration groups in the TCGA-KIRC dataset and the FerrDb V2 database. Using univariate and multivariate Cox regression, we identified two independent prognostic genes, AMN and PDK4, and constructed an IRFG score model immune-related ferroptosis genes risk score (IRFGRs) to evaluate its prognostic value in ccRCC. The IRFGRs demonstrated excellent and stable performance for predicting ccRCC patient survival in both the TCGA training set and the ArrayExpress validation set, with an AUC range of 0.690-0.754, outperforming other commonly used clinicopathological indicators. Our findings enhance the understanding of TME infiltration with ferroptosis and identify immune-mediated ferroptosis genes associated with prognosis in ccRCC.

A rapid and highly sensitive multiple detection of human adenovirus type 3, type 7 and respiratory syncytial virus by recombinase-aided reverse transcription PCR.

BACKGROUND: Polymerase chain reaction (PCR) has been widely used for many pathogen detection. However, PCR technology still suffers from long detection time and insufficient sensitivity. Recombinase-aided amplification (RAA) is a powerful nucleic acid detection tool with high sensitivity and amplification efficiency, but its complex probes and inability of multiplex detection hinder the further application of this technology. METHODS: In this study, we developed and validated the multiplex reverse transcription recombinase-aided PCR (multiplex RT-RAP) assay for human adenovirus 3 (HADV3), human adenovirus 7 (HADV7), and human respiratory syncytial virus (HRSV) within 1 h with Human RNaseP protein as a reference gene to monitor the whole process. RESULTS: Using recombinant plasmids, the sensitivity of multiplex RT-RAP for the detection of HADV3, HADV7, and HRSV was 18, 3, and 18 copies per reaction, respectively. The multiplex RT-RAP showed no cross-reactivity with other respiratory viruses, demonstrating its good specificity. A total of 252 clinical specimens were tested by multiplex RT-RAP and the results were found to be consistent with those of corresponding RT-qPCR assays. After testing serial dilutions of selected positive specimens, the detection sensitivity of multiplex RT-RAP was two to eightfold higher than that of corresponding RT-qPCR. CONCLUSION: We conclude the multiplex RT-RAP is a robust, rapid, highly sensitive, and specific assay with the potential to be used in the screening of clinical samples with low viral load.

Sports knee joint injury in ski training and its prevention / Lesão esportiva da articulação do joelho em treino de esqui e sua prevenção / Lesión de la articulación de la rodilla en el entrenamiento de esquí y su prevención

ABSTRACT Introduction Ski training presents the high ability of athletes and strong demand on the physical conditioning of the knee joint, which is frequently injured. Objective Study the sports injuries of the knee joint in ski training and explore its preventive measures. Methods The interview method was used with ski instructors, enthusiasts, and athletes by questionnaire with design and distribution of topics related to the occurrence of injuries. Sport injury factors are discussed internally and externally. Preventive measures and protective equipment for knee sports injuries are presented. Results Currently, the proportion of serious sports injuries to the knee joint is relatively high, and problems such as collision are the most common injury factors, and differences exist between male and female athletes. Among the causes of injury, the highest-scoring technical factor for injury was "deviation from technical movements", and the highest-scoring preventive measures were "strength training for vulnerable parts" and "sufficient preparation for activities". Conclusion The instructors should provide effective training programs according to the actual situation of the athletes, pay attention to monitoring the skiing environment, and recommend the appropriate protective equipment for the sport. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução O treino de esqui apresenta alta habilidade dos atletas e forte demanda no condicionamento físico da articulação do joelho, que é frequentemente lesionada. Objetivo Estudar as lesões esportivas da articulação do joelho no treino de esqui e explorar suas medidas preventivas. Métodos O método de entrevista foi usado com instrutores, entusiastas e esportistas de esqui por questionário com projetação e distribuição de tópicos relacionados à ocorrência das lesões. Os fatores de lesão esportiva são discutidos interna e externamente. Medidas preventivas e equipamentos de proteção para lesões esportivas no joelho são apresentados. Resultados Atualmente, a proporção de lesões esportivas graves na articulação do joelho é relativamente alta e problemas como colisão são os fatores de lesão mais comuns, existindo diferenças entre atletas do sexo masculino e feminino. Entre as causas de lesão, o fator técnico de maior pontuação para lesão foi "desvio de movimentos técnicos", e as medidas preventivas de maior pontuação foram "treinamento de força para partes vulneráveis" e "preparação suficiente para as atividades". Conclusão Os treinadores devem fornecer programas de treino eficazes de acordo com a situação real dos atletas, atentarem-se ao monitoramento do ambiente de esqui e indicar os equipamentos de proteção adequados ao esporte. Nível de evidência II; Estudos terapêuticos - investigação dos resultados do tratamento.

RESUMEN Introducción El entrenamiento de esquí presenta una alta capacidad de los deportistas y una fuerte exigencia en el acondicionamiento físico de la articulación de la rodilla, que se lesiona con frecuencia. Objetivo Estudiar las lesiones deportivas de la articulación de la rodilla en el entrenamiento de esquí y explorar sus medidas preventivas. Métodos Se utilizó el método de la entrevista con instructores de esquí, aficionados y deportistas mediante un cuestionario con diseño y distribución de temas relacionados con la ocurrencia de las lesiones. Los factores de las lesiones deportivas se discuten interna y externamente. Se presentan las medidas preventivas y los equipos de protección para las lesiones deportivas de rodilla. Resultados En la actualidad, la proporción de lesiones deportivas graves en la articulación de la rodilla es relativamente alta y problemas como la colisión son los factores de lesión más comunes, y existen diferencias entre los deportistas masculinos y femeninos. Entre las causas de las lesiones, el factor técnico con mayor puntuación fue la "desviación de los movimientos técnicos", y las medidas preventivas con mayor puntuación fueron el "entrenamiento de fuerza de las partes vulnerables" y la "preparación suficiente para las actividades". Conclusión Los entrenadores deben proporcionar programas de entrenamiento eficaces en función de la situación real de los deportistas, prestar atención al seguimiento del entorno de esquí e indicar el equipo de protección adecuado para el deporte. Nivel de evidencia II; Estudios terapéuticos - investigación de los resultados del tratamiento.

Analysis of the effect of high intensity intermittent training for young athletes / Análise do efeito do treinamento intermitente de alta intensidade para jovens atletas / Análisis del efecto del entrenamiento intermitente de alta intensidad en atletas jóvenes

ABSTRACT Introduction The CPC Central Committee and the State Council have pointed out that currently, the basic indicators of young students' physical health have completely declined. This circumstance has made adolescent physical health and improving the quality of young athletes a key issue in physical education targeting adolescents. Objective Investigate the effect of high-intensity intermittent training on young athletes. Methods 24 adolescents performed 10 intermittent high-intensity training for one month, and all athletes' pre and post-training indexes were collected and compared. Results The physical examination results of male and female athletes improved to some extent (athletes' blood pressure fluctuated during the 10 training sessions, increasing slightly, and heart rate variability increased considerably (P < 0.05). Conclusion High-intensity intermittent training has a good effect on health promotion in young athletes, and can effectively improve their physical function and competitive performance. Physical education teachers and student-athletes should conduct relevant training. In addition, this training method also fits the current learning life situation of high school students. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução O Comitê Central do CPC e o Conselho de Estado destacaram que, atualmente, os indicadores básicos da saúde física dos jovens estudantes têm diminuído de forma completa. Essa circunstância tornou a saúde física dos adolescentes e a melhoria da qualidade dos jovens atletas um ponto-chave na educação física direcionada aos adolescentes. Objetivo Investigar o efeito do treinamento intermitente de alta intensidade em jovens atletas. Métodos 24 adolescentes realizaram 10 treinos intermitentes de alta intensidade, com duração de um mês, todos os índices pré e pós-treino dos atletas foram coletados e comparados. Resultados Os resultados do exame físico de atletas do sexo masculino e feminino melhoraram em certa medida (a pressão arterial dos atletas flutuou durante as 10 sessões de treinamento, aumentando ligeiramente, e a variabilidade da frequência cardíaca aumentou consideravelmente (P < 0,05). Conclusão O treinamento intermitente de alta intensidade tem um bom efeito na promoção da saúde nos jovens atletas, podendo melhorar efetivamente sua função física e desempenho competitivo. Professores de educação física e atletas estudantes devem realizar treinamentos relevantes. Além disso, esse método de treinamento também se encaixa na situação atual da vida de aprendizagem dos alunos do ensino médio. Nível de evidência II; Estudos terapêuticos - investigação dos desfechos do tratamento.

RESUMEN Introducción El Comité Central del CPC y el Consejo de Estado han puesto de relieve que en la actualidad los indicadores básicos de salud física de los jóvenes estudiantes han disminuido completamente. Esta circunstancia hizo que la salud física de los adolescentes y la mejora de la calidad de los jóvenes deportistas fuera un punto clave en la educación física dirigida a los adolescentes. Objetivo Investigar el efecto del entrenamiento intermitente de alta intensidad en atletas jóvenes. Métodos 24 adolescentes realizaron 10 sesiones de entrenamiento intermitente de alta intensidad de un mes de duración, se recogieron y compararon los índices de todos los atletas antes y después del entrenamiento. Resultados Los resultados de la exploración física de los atletas masculinos y femeninos mejoraron en cierta medida (la presión arterial de los atletas fluctuó durante las 10 sesiones de entrenamiento, aumentando ligeramente, y la variabilidad de la frecuencia cardíaca aumentó considerablemente (P < 0,05). Conclusión El entrenamiento intermitente de alta intensidad tiene un buen efecto en la promoción de la salud en los atletas jóvenes, y puede mejorar eficazmente su función física y su rendimiento competitivo. Los profesores de educación física y los alumnos deportistas deben realizar la formación pertinente. Además, este método de formación también se ajusta a la situación actual de la vida de aprendizaje de los estudiantes de secundaria. Nivel de evidencia II; Estudios terapêuticos - investigación de los resultados del tratamiento.

Development of field-applicable endogenous internally controlled recombinase-aided amplification (EIC-RAA) assays for the detection of human papillomavirus genotypes 6 and 11 using sample releasing agent.

Objective: Human papillomavirus (HPV) 6 and 11 are the two most common low-risk HPV subtypes, accounting for more than 90% of condyloma acuminatum. A simple, accurate and rapid screening method to be applied in community-level hospitals is in high demand. Methods: Endogenous internally controlled recombinase-assisted amplification (EIC-RAA) assays for HPV6 and 11 were performed in a single closed-tube at 39 °C within 30 min. The sensitivity and specificity of EIC-RAA were examined using recombinant plasmids and pre-tested HPV DNA. A total of 233 clinical samples were collected, and the DNA was extracted by traditional multi-step extraction, or sample releasing agent, before analysis by EIC-RAA. For comparison, HPV detection via Quantitative real-time PCR (qPCR) was also performed. Results: The sensitivity of EIC-RAA analysis was 10 copies/reaction for HPV6, 100 copies/reaction for HPV11, and 100 copies/reaction for the human ß-globin gene. No cross-reaction was observed with other HPV subtypes. Clinical performance of the EIC-RAA assay achieved a 100% of concordance rate with the commercial HPV qPCR kit. Further, the EIC-RAA assay achieved a 100% of concordance rate when using multi-step extracted DNA and sample releasing agent-processed DNA. Summary: The EIC-RAA assay for HPV6 and 11 detection possesses the advantages of accuracy, simplicity and rapidity, and demonstrates great potential to be used in community-level hospitals for field investigation.

Combination of Androgen Deprivation Therapy with Radical Local Therapy Versus Androgen Deprivation Therapy Alone for Newly Diagnosed Oligometastatic Prostate Cancer: A Phase II Randomized Controlled Trial.

BACKGROUND: Previous studies suggested that men with metastatic prostate cancer might benefit from local treatment of the primary tumor. OBJECTIVE: To determine whether radical local therapy (RLT) improves survival for men with oligometastatic prostate cancer (OMPCa). DESIGN, SETTING, AND PARTICIPANTS: This open-label randomized controlled trial included patients with newly diagnosed OMPCa defined as five or fewer bone or extrapelvic lymph node metastases and no visceral metastases. INTERVENTION: Patients were randomly allocated to androgen deprivation therapy (ADT) or ADT and RLT. Men allocated RLT received either cytoreductive radical prostatectomy (RP) or prostate radiation therapy (RT) with a radical dose schedule. OUTCOME MEASUREMENTS AND STATISTICAL ANALYSIS: The primary outcome was radiographic progression-free survival (rPFS). Secondary outcomes were overall survival (OS) and prostate-specific antigen (PSA) progression-free survival. RESULTS AND LIMITATIONS: Between September 2015 and March 2019, 200 patients were randomized, with 100 men allocated to each group. The median age was 68 yr and the median PSA at diagnosis was 99 ng/ml. In the study group, 96 patients underwent RLT (85 RP and 11 RT). In the control group, 17 patients eventually received RLT (15 RP and two RT). All patients were included for an intention-to-treat analysis. After a median follow-up of -48 mo, the median rPFS was not reached in the study group and was 40 mo in the control group (hazard ratio [HR] 0.43, 95% confidence interval [CI] 0.27-0.70; p = 0.001). The 3-yr OS rate was 88% for the study group and 70% for the control group (HR 0.44, 95% CI 0.24-0.81; p = 0.008). CONCLUSIONS: Men with newly diagnosed OMPCa who received ADT plus RLT (mainly prostatectomy) had significantly higher rates of rPFS and OS than those who received ADT alone. PATIENT SUMMARY: This study investigated the effect of radical local therapy (RLT) of the primary tumor on survival in patents with oligometastatic prostate cancer. In our group, RLT improved radiographic progression-free and overall survival.

Aetiological Characteristics of Infectious Diarrhea in Yantai City, Shandong Province, China in 2017.

This study aimed to analyse the pathogenic spectrum and epidemiological characteristics of infectious diarrhea in Yantai City, Shandong Province, China and provide a reference for its prevention and control. A total of 713 stool specimens collected within 3 days of diarrhea onset from January to December 2017 at secondary or higher hospitals in Yantai City were tested for 10 causative pathogens, using real-time polymerase chain reaction (RT-PCR). The top two rotaviruses and norovirus were analysed for typing and geographical distribution. The total positive rate was 46.56% (332/713), and 268 of 713 specimens contained at least one pathogen; 64 had at least two pathogens, accounting for 19.28% of the positive specimens (64/332). The positivity rates of rotavirus (RV), norovirus (NoVs) GI, norovirus (NoVs) GII, enterovirus universal (EV), enteric adenoviruses (EAdV), sapovirus (SaV), astrovirus (Astv), Salmonella (SE), Listeria monocytogenes (LiMo), and Vibrio parahaemolyticus (VP) were 20.06% (143/713), 1.82% (13/713), 12.84% (89/713), 10.66% (76/713), 4.07% (29/713), 0.42% (3/713), 2.38% (17/713), 1.54% (11/713), 1.82% (13/713), and 1.54% (11/713), respectively. Infectious diarrhea showed a high prevalence in young children aged 1-5 years, accounting for 48.6% of the total number of cases. Bacterial diarrhea was predominant in summer, and viral diarrhea was distributed throughout the year, without a significant seasonal pattern. Rotavirus is dominated by G9P, accounting for 81.82%, while norovirus is dominated by the GII type and has diverse characteristics. The aetiology of infectious diarrhea in Yantai is mainly viral, with RV, NoVs, EV, EAdV, and Astv being the most frequent pathogens. Continuous surveillance of infectious diarrhea diseases can help us understand its epidemiological and pathogenic characteristics, thereby taking targeted preventive and control measures in different seasons.

Does the primary tumor site in stage I extranodal natural killer/T-cell lymphoma matter?

The purpose of this study was to investigate whether the primary tumor site in stage I extranodal natural killer/T-cell lymphoma (ENKTCL) had a prognostic value. Between January 2009 and December 2015, 152 stage I ENKTCL patients with primary disease in the nasal cavity and Waldeyer's ring were enrolled for this retrospective study. All patients received extended field intensity-modulated radiotherapy alone without prophylactic cervical node irradiation at a total dose of 50 Gy. In this study, there were 122 patients whose primary tumors were localized in the nasal cavity (NC group), and no adjacent structures were involved. A total of 18 patients had a primary disease involving the nasal cavity and Waldeyer's ring (NC-WR group), and the remaining 12 patients had primary tumors confined to Waldeyer's ring (WR group). We found that there was no significant difference in cervical lymph node failure rates among the NC, NC-WR, and WR groups. In terms of the 5-year overall survival (OS) rates, there was a significant difference among the NC, NC-WR, and WR groups (p=0.004), with the WR group having the worst OS. Multivariate analyses showed that the primary site (p=0.011) and ECOG (Eastern Cooperative Oncology Group) score (p=0.013) were independent prognostic factors for OS. In summary, patients with stage I ENKTCL had a good local control rate with radiotherapy alone and without prophylactic cervical node irradiation (PCNI), regardless of the site of the primary tumor. So, we think PCNI for stage I ENKTCL patients is not necessary. Patients with a primary tumor site located in Waldeyer's ring had the worst prognosis. And combined treatment with radiotherapy and chemotherapy should be considered in patients with primary tumors located outside the nasal cavity.

Enhancement of a recombinase-aided amplification assay using betaine and pullulan.

Background: Nucleic acid amplification enhancers suitable for use in a recombinase-aided amplification (RAA) assay were studied for the first time, and amplification of a long-fragment (509 bp) was initially explored. Methods: Using recombinant plasmids and clinical samples, RAA fluorescence and basic methods were used to evaluate the efficacy. The fluorescence method was evaluated by threshold time and fluorescence value, and the basic method was characterized by 2% agarose gel electrophoresis. Results: Taking a previously established RAA assay for HPV18 as an example, we demonstrated that the addition of 0.2 M, 0.4 M, and 0.6 M betaine and 10% pullulan could enhance the RAA. The new RAA assays with betaine and pullulan were named B-RAA and P-RAA, respectively. Using the B-RAA and P-RAA fluorescence methods, the threshold time values could be shortened by 1.72-2.32 minutes and 2.60 minutes, respectively, and the fluorescence values could be enhanced by 8847.25-9094.37 mv and 5250 mv, respectively. Using the basic method, the sensitivity could be increased 10-fold. We successfully amplified a long-fragment of 509 bp using a P-RAA assay with a sensitivity of 102 copies/µL (compared with 103 copies/µL in the RAA assay). Conclusions: Thus, we concluded that betaine and pullulan are effective additives to enhance the sensitivity of RAA assays.

RAP: A Novel Approach to the Rapid and Highly Sensitive Detection of Respiratory Viruses.

Recombinase aided amplification (RAA) is an emerging isothermal amplification method used for detecting various pathogens. However, RAA requires a complex and long probe to ensure high sensitivity during fluorescence assay. TaqMan probe used for quantitative PCR (qPCR) is simple and universal. Herein, we developed a new approach for detecting nucleic acids of pathogens, known as RAP (Recombinase aided PCR). The method combines RAA and qPCR to ensure a rapid and highly sensitive detection using a conventional qPCR device. RAP is a two-stage amplification process performed in a single tube within 1 hour. The method involves an RAA reaction for 10 min at 39°C (first stage) followed by 15 cycles of qPCR (second stage). Using human adenovirus 3 (HADV3) and human adenovirus 7 (HADV7) plasmids, the sensitivities of RAP assays for detecting HADV3 and HADV7 were 6 and 17 copies per reaction, respectively. The limit of RAP detection was at least 16-fold lower than the corresponding qPCR, and no-cross reaction with other respiratory viruses was observed. The results of RAP analysis revealed 100% consistency with qPCR assay. This study shows that RAP assay is a rapid, specific, and highly sensitive detection method with a potential for clinical and laboratory application.

Phase II study of sequential chemoradiotherapy with L-asparaginase, dexamethasone, ifosfamide, cisplatin, and etoposide (DICE-L) in the early stage of extranodal natural killer (NK)/T-cell lymphoma.

BACKGROUND: To explore a more effective treatment strategy for newly diagnosed stage I and II extranodal natural killer/T-cell lymphoma (ENKTL), nasal type, we conducted a prospective phase II study of sequential chemoradiotherapy with the L-asparaginase, dexamethasone, ifosfamide, cisplatin, and etoposide (DICE-L) regimen. METHODS: Patients with newly diagnosed stage I and II ENKTL in the upper-aerodigestive tract were enrolled. Treatment was comprised of up to 4 cycles of DICE-L followed by 50 Gy of intensity modulated radiation therapy (IMRT) to the involved field. The primary endpoint was the complete response (CR) rate. The secondary endpoints were the objective response rate (ORR), the 5-year overall survival (OS) rate, the 5-year progression-free survival (PFS) rate, and safety. RESULTS: A total of 81 patients were enrolled from June 2009 to May 2012 in Shanghai Cancer Hospital. Among these patients, 68 patients achieved CR and 1 patient achieved partial response (PR). The CR rate was 84%, and the ORR was 85.2%. With a median follow up of 88.1 months, the 5-year OS and 5-year PFS rates were 82.4% and 63.4%, respectively. The most common adverse events were grade 3 to 4 neutropenia (73.5%) and febrile neutropenia (21%). CONCLUSIONS: Sequential chemoradiotherapy using DICE-L followed by radiotherapy is an effective treatment modality for stage I to IIE ENKTL and is safe with acceptable toxicity.

Sports mechanics rehabilitation effects on knee and muscle / Os efeitos da reabilitação mecânica do esporte sobre o joelho e o músculo / Los efectos de la rehabilitación mecánica del deporte sobre la rodilla y el músculo

ABSTRACT Introduction: The knee joint is a complex and important joint in the human body. It plays an active role in maintaining posture balance of the human body in sports, allowing one to bear a larger load and complete various complex technical movements. Objective: To investigate the stability and influence of kinemechanics in knee joint rehabilitation training. Methods: A total of 80 patients with anterior cruciate ligament injury who received treatment in Henan Provincial People's Hospital were selected and divided into control group and observation group according to the random number table method, with 40 cases in each group. Results: Six months after surgery, the peak torque (PT), peak torque to body weight ratio (PT/BWT), total work (TWT), Lysholm score and Tegner score, and knee circumference of the two groups were significantly improved compared with those before surgery (P <0.01). The degree of improvement was greater in the observation group:compared with the control group, the difference was highly statistically significant (P<0.01). Conclusions: Professional exercise rehabilitation has a definite effect on patients after anterior cruciate ligament reconstruction. It can significantly improve the muscle strength of quadriceps and hamstring, improve the function of knee joint and promote its recovery. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução: A junta do joelho é uma junta complexa e importante para o corpo humano. Tem um papel ativo na manutenção do equilíbrio postural do corpo humano nos esportes, permitindo que se suporte cargas maiores e que se realize vários movimentos técnicos complexos. Objetivo: Investigar a estabilidade e influência da cinemática no treino para reabilitação da junta do joelho. Métodos: Um total de 80 pacientes com lesão de ligamento cruzado anterior que receberam tratamento no Hospital do Povo da Província de Henan foram selecionados e divididos em grupo de controle e grupo de observação, de acordo com o método de tabela numérica aleatória, com 40 casos em cada grupo. Resultados: Seis meses após a cirurgia, o torque de pico (TP), a proporção torque de pico-peso corporal (TP-PC), trabalho total (TT), escore Lysholm e escore Tegner, e a circunferência do joelho dos dois grupos melhoraram significativamente em relação à antes da cirurgia (P <0.01). O grau de recuperação foi maior no grupo de observação: comparado com o grupo de controle, a diferença é estatisticamente significativa (P <0.01). Conclusões: A reabilitação com exercícios profissionais definitivamente tem efeito em pacientes após a reconstrução do ligamento cruzado anterior, pois pode aumentar a força muscular de quadríceps e tendões, melhorar a função da junta do joelho e levar a sua recuperação. Nível de evidência II; estudos terapêuticos - investigação de resultados de tratamento.

RESUMEN Introducción: La articulación de la rodilla es compleja e importante para el cuerpo humano. Tiene un papel activo en la manutención del equilibrio postural del cuerpo humano en los deportes, permitiendo que se soporte cargas mayores y que se realice varios movimientos técnicos complejos. Objetivo: Investigar la estabilidad e influencia de la cinemática en el entrenamiento para rehabilitación de la articulación de la rodilla. Métodos: Se seleccionó un total de 80 pacientes con lesión de ligamento cruzado anterior que recibieron tratamiento en el Hospital del Pueblo de la Provincia de Henan y se los dividió en grupo de control y grupo de observación, de acuerdo con el método de tabla de números aleatorios, con 40 casos en cada grupo. Resultados: Seis meses tras la cirugía, el torque de pico (TP), la proporción torque de pico-peso corporal (TP-PC), trabajo total (TT), puntuación Lysholm y puntuación Tegner, y la circunferencia de la rodilla de los dos grupos mejoraron significativamente en relación a antes de la cirugía (P<0.01). El grado de recuperación fue mayor en el grupo de observación: comparado con el grupo de control, la diferencia es estadísticamente significativa (P<0.01). Conclusiones: La rehabilitación con ejercicios profesionales definitivamente tiene efecto en pacientes tras la reconstrucción del ligamento cruzado anterior, pues puede aumentar la fuerza muscular de cuádriceps y tendones, mejorar la función de la articulación de la rodilla y llevar a su recuperación. Nivel de evidencia II; Estudios terapéuticos - investigación de resultados de tratamiento.

The prevention method of sports medicine imaging modeling / O método preventivo da medicina do esporte no modelo de imagens / El método preventivo de la medicina del deporte en el modelo de imágenes

ABSTRACT Introduction: Sports injury is an important reason to interfere with sports training and physical exercise since it will not only bring unnecessary pain to the injured body, but also bring out negative emotions in athletes. Objective: To study the sports injury characteristics of high level competitive aerobics athletes in China. Methods: Literature data, questionnaire survey, expert interviews, field observation, mathematical statistics and physiotherapy tracking were used. Results: Wrists, ankles, waist and knees were the most frequently injured parts in high level competitive aerobics athletes in China. The main type of injury was acute injury, but chronic injury (48.18%) could not be ignored. Ninety percent of active high-level competitive aerobics athletes train with injuries, and 10 percent of them completely stop training because of injuries. Conclusions: Athletes should pay attention to their physical reserve and strengthen physical training. They must also be attentive to self-supervision, their own subjective feelings, and control the amount of exercise done. Level of evidence II; Therapeutic studies - investigation of treatment results.

RESUMO Introdução: As lesões no esporte são uma razão importante para interferir no treinamento esportivo e no exercício físico, já que não apenas levam a dor desnecessária do corpo lesado, mas também levam a emoções negativas em atletas. Objetivo: Estudar as características de lesões esportivas de atletas de ginástica aeróbica de alto nível competitivo na China. Métodos: Dados da literatura, questionário de sondagem, entrevistas especializadas, observação de campo, estatísticas matemáticas e acompanhamento fisioterapêutico foram usados. Resultados: Pulsos, tornozelos, cintura e joelhos são as partes lesadas com maior frequência nas atletas de ginástica aeróbica de alto nível competitivo na China. O tipo mais comum é a lesão aguda, porém a lesão crônica (48,18%) não pode ser ignorada. Noventa por cento de atletas de ginástica aeróbica de alto nível competitivo na ativa treinam com lesões, e 10% param de treinar completamente devido às lesões. Conclusões: Atletas devem ficar atentos a suas reservas fiscais e fortalecer seu treino físico. Devem também ficar atentos a auto supervisão, ao seu estado emocional e controlar a quantidade exercícios que praticam. Nível de evidência II; Estudos terapêuticos - investigação de resultados de tratamento.

RESUMEN Introducción: Las lesiones en el deporte son una razón importante para interferir en el entrenamiento deportivo y en el ejercicio físico, ya que no solo llevan al dolor desnecesario del cuerpo lesionado, sino que llevan a emociones negativas en atletas. Objetivo: Estudiar las características de lesiones deportivas de atletas de gimnasia aeróbica de alto nivel competitivo en China. Métodos: Se utilizaron datos de la literatura, encuestas de sondeo, entrevistas especializadas, observación de campo, estadísticas matemáticas y acompañamiento fisioterapéutico. Resultados: Muñecas, tobillos, cintura y rodillas son las partes lesionadas con mayor frecuencia en las atletas de gimnasia aeróbica de alto nivel competitivo en China. El tipo más común es la lesión aguda, pero la lesión crónica (48,18%) no puede ignorarse. Noventa por ciento de atletas de gimnasia aeróbica de alto nivel competitivo en China entrenan con lesiones, y 10% paran de entrenar completamiento debido a las lesiones. Conclusiones: Atletas deben atentarse a sus reservas fiscales y fortalecer su entrenamiento físico. Deben también atentarse a la autosupervisión, a su estado emocional y controlar la cantidad de ejercicios que practican. Nivel de evidencia II; Estudios terapéuticos - investigación de resultados de tratamiento.

A duplex probe-directed recombinase amplification assay for detection of single nucleotide polymorphisms on 8q24 associated with prostate cancer

Single nucleotide polymorphisms (SNPs) have important application value in the research of population genetics, hereditary diseases, tumors, and drug development. Conventional methods for detecting SNPs are typically based on PCR or DNA sequencing, which is time-consuming, costly, and requires complex instrumentation. In this study, we present a duplex probe-directed recombinase amplification (duplex-PDRA) assay that can perform real-time detection of two SNPs (rs6983267 and rs1447295) in four reactions in two tubes at 39°C within 30 min. The sensitivity of duplex-PDRA was 2×103-104 copies per reaction and no cross-reactivity was observed. A total of 382 clinical samples (179 prostate cancer patients and 203 controls) from northern China were collected and tested by duplex-PDRA assay and direct sequencing. The genotyping results were completely identical. In addition, the association analysis of two SNPs with prostate cancer risk and bone metastasis was conducted. We found that the TT genotype of rs6983267 (OR: 0.42; 95%CI: 0.23-0.78; P=0.005) decreased the risk of prostate cancer, while the CA genotype of rs1447295 (OR: 1.89; 95%CI: 1.20-2.96; P=0.005) increased the risk of prostate cancer. However, no association between the two SNPs (rs6983267 and rs1447295) and bone metastasis in prostate cancer was found in this study (P>0.05). In conclusion, the duplex-PDRA assay is an effective method for the simultaneous detection of two SNPs and shows great potential for widespread use in research and clinical settings.

A duplex probe-directed recombinase amplification assay for detection of single nucleotide polymorphisms on 8q24 associated with prostate cancer.

Single nucleotide polymorphisms (SNPs) have important application value in the research of population genetics, hereditary diseases, tumors, and drug development. Conventional methods for detecting SNPs are typically based on PCR or DNA sequencing, which is time-consuming, costly, and requires complex instrumentation. In this study, we present a duplex probe-directed recombinase amplification (duplex-PDRA) assay that can perform real-time detection of two SNPs (rs6983267 and rs1447295) in four reactions in two tubes at 39°C within 30 min. The sensitivity of duplex-PDRA was 2×103-104 copies per reaction and no cross-reactivity was observed. A total of 382 clinical samples (179 prostate cancer patients and 203 controls) from northern China were collected and tested by duplex-PDRA assay and direct sequencing. The genotyping results were completely identical. In addition, the association analysis of two SNPs with prostate cancer risk and bone metastasis was conducted. We found that the TT genotype of rs6983267 (OR: 0.42; 95%CI: 0.23-0.78; P=0.005) decreased the risk of prostate cancer, while the CA genotype of rs1447295 (OR: 1.89; 95%CI: 1.20-2.96; P=0.005) increased the risk of prostate cancer. However, no association between the two SNPs (rs6983267 and rs1447295) and bone metastasis in prostate cancer was found in this study (P>0.05). In conclusion, the duplex-PDRA assay is an effective method for the simultaneous detection of two SNPs and shows great potential for widespread use in research and clinical settings.

Baseline SUVmax of 18F-FDG PET-CT indicates prognosis of extranodal natural killer/T-cell lymphoma.

To evaluate the prognostic value of the baseline SUVmax of F-FDG PET-CT in extranodal natural killer/T-cell lymphoma (NKTCL) patients.From January 2010 to December 2015, 141 extranodal NKTCL patients with staging F-FDG PET-CT scan were divided into two group based on SUVmax cutoff value obtained from operating characteristic (ROC) curves. All the patients received radiotherapy, chemotherapy or chemoradiation. Survival analysis was performed on the basis of SUVmax.The median baseline SUVmax of the tumors was 11.67 (range 2.6-34.6). The ROC curves showed that the optimal cutoff of the baseline SUVmax was 9.65. The patients were divided into two groups: low SUV group (SUVmax < 9.65) and high SUV group (SUVmax ≥ 9.65). Patients in high SUV group were more likely to have invasive disease outside the nasal cavity (P < .001), poorer ECOG scores (P = .012) and higher LDH levels (P = .034). The univariate survival analyses indicated that high SUVmax was a poor prognostic factor for overall survival (OS, P = .038), progression free survival (PFS, P = .006) and distant relapse free survival (DRFS, P = .001), but not for local recurrence free survival (LRFS, P > .05). These results were consistent with that of the survival analyses using the Kaplan-Meier method. The multivariate survival analyses showed that the baseline SUVmax was no longer a prognostic factor for OS (HR 1.99, 95% CI 0.81-4.88, P = .135), but it still indicated worse PFS (HR 2.6, 95% CI 1.24-5.46, P = .012) and DRFS (HR 4.58, 95% CI 1.83-11.46, P = .001) independent of other variables.For extranodal NKTCL patients, a higher baseline SUVmax of F-FDG PET-CT was associated with more aggressive clinical features. An SUVmax ≥ 9.65 was an independent poor prognostic factor for DRFS and PFS. Thus, the baseline SUVmax may be a valuable tool to help identify patients with a high risk of disease progression.

Internally controlled recombinase-aided amplification (IC-RAA) assays for the detection of human papillomavirus genotypes 16 and 18 using extracted DNA and samples treated with nucleic acid releasing agent.

Cervical cancer is primarily caused by persistent infection with high-risk human papillomavirus (HPV), and 70% of cases are associated with HPV16 and 18 infections. The objective of this study was to establish rapid, simple, and sensitive internally controlled recombinase-aided amplification (IC-RAA) assays for the detection of HPV16 and 18. The assays were performed at 39 ℃ and were completed within 30 min. A total of 277 clinical samples of exfoliated cervical cells were tested by IC-RAA assays and commercial HPV real-time fluorescent PCR kits using extracted DNA and samples treated with nucleic acid releasing agent. The analytical sensitivity of the IC-RAA assay was found to be 10 copies/µL for the detection of HPV16 and 18 when using recombinant plasmids as targets. The optimal concentration of the internal control (IC) plasmid and 18 was 1000 copies/µL for HPV16 and 100 copies/µL for HPV18. The clinical sensitivity of the IC-RAA assays for HPV16 using extracted DNA and samples treated with nucleic acid releasing agent was 98.73% and 97.47%, respectively, with kappa values of 0.977 (P < 0.01) and 0.955 (P < 0.01), respectively, and 100% The specificity in both cases. For HPV18, the sensitivity and specificity were 100%, and the kappa value was 1 for both samples (P < 0.01). The IC-RAA assay is a promising tool for the detection of HPV16 and HPV18, especially in resource-constrained settings.