Search

Sijin Wu,1 Zhongli Chen,1 Yuan Gao,1 Songren Shu,2 Feng Chen,1 Ying Wu,1 Yan Dai,1 Shu Zhang,1 Keping Chen1 1Arrhythmia Center, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, People’s Republic of China; 2Department of Cardiovascular Surgery, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, People’s Republic of ChinaCorrespondence: Keping Chen, Arrhythmia Center, Fuwai Hospital, National Center for Cardiovascular Diseases, Chinese Academy of Medical Sciences & Peking Union Medical College, 167 Beilishi Road, Xicheng District, Beijing, 100037, People’s Republic of China, Email chenkeping@263.netBackground: The diagnosis of cardiac syncope remains a challenge. This study sought to develop and validate a diagnostic model for the early identification of individuals likely to have a cardiac cause.Methods: 877 syncope patients with a determined cause were retrospectively enrolled at a tertiary heart center. They were randomly divided into the training set and validation set at a 7:3 ratio. We analyzed the demographic information, medical history, laboratory tests, electrocardiogram, and echocardiogram by the least absolute shrinkage and selection operator (LASSO) regression for selection of key features. Then a multivariable logistic regression analysis was performed to identify independent predictors and construct a diagnostic model. The receiver operating characteristic curves, area under the curve (AUC), calibration curves, and decision curve analysis were used to evaluate the predictive accuracy and clinical value of this nomogram.Results: Five independent predictors for cardiac syncope were selected: BMI (OR 1.088; 95% CI 1.022– 1.158; P =0.008), chest symptoms preceding syncope (OR 5.251; 95% CI 3.326– 8.288; P < 0.001), logarithmic NT-proBNP (OR 1.463; 95% CI 1.240– 1.727; P < 0.001), left ventricular ejection fraction (OR 0.940; 95% CI 0.908– 0.973; P < 0.001), and abnormal electrocardiogram (OR 6.171; 95% CI 3.966– 9.600; P < 0.001). Subsequently, a nomogram based on a multivariate logistic regression model was developed and validated, yielding AUC of 0.873 (95% CI 0.845– 0.902) and 0.856 (95% CI 0.809– 0.903), respectively. The calibration curves showcased the nomogram’s reasonable calibration, and the decision curve analysis demonstrated good clinical utility.Conclusion: A diagnostic tool providing individualized probability predictions for cardiac syncope was developed and validated, which may potentially serve as an effective tool to facilitate early identification of such patients.Keywords: cardiac syncope, syncope, diagnosis, nomogram, prediction model

Caiyun Wang,1 Hongmei Xu,2 Gang Liu,3,4 Jing Liu,5 Hui Yu,6 Biquan Chen,7 Guo Zheng,8 Min Shu,9 Lijun Du,10 Zhiwei Xu,11 Lisu Huang,1,12 Haibo Li,13 Sainan Shu,14 Yinghu Chen,1 Corporate Authors of Chinese Pediatric Bacterial Meningitis Surveillance (CPBMS) Study Group:Dong Wang, Huiling Deng, Songting Bai, Qingwen Shan, Chunhui Zhu, Jianmei Tian, Jianhua Hao, Aiwei Lin, Daojiong Lin, Jinzhun Wu, Xinhua Zhang, Qing Cao, Zhongbin Tao, Yuan Chen, Guolong Zhu, Ping Xue, Zhengzhen Tang, Xuewen Su, Zhenghai Qu, Shiyong Zhao, Lin Pang On behalf of The CPBMS Study Group1Department of Infectious Disease, Children’s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, National Children’s Regional Medical Center, Hangzhou, Zhejiang, People’s Republic of China; 2Department of Infectious Disease, Children’s Hospital of Chongqing Medical University, Chongqing, People’s Republic of China; 3Department of Infectious Diseases, Key Laboratory of Major Diseases in Children, Ministry of Education, Beijing Children’s Hospital, Capital Medical University, National Center for Children’s Health, Beijing, People’s Republic of China; 4Research Unit of Critical Infection in Children, Chinese Academy of Medical Sciences, Beijing, People’s Republic of China; 5Department of Infectious Disease, Hunan Children’s Hospital, Changsha, Hunan, People’s Republic of China; 6Department of Infectious Disease, The Children’s Hospital of Fudan University, Shanghai, People’s Republic of China; 7Department of Infection, Anhui Province Children’s Hospital, Hefei, Anhui, People’s Republic of China; 8Department of Neurology, Children’s Hospital of Nanjing Medical University, Nanjing, Jiangsu, People’s Republic of China; 9Department of Pediatrics, West China Second University Hospital, Sichuan University/ West China Women’s and Children’s Hospital, Chengdu, Sichuang, People’s Republic of China; 10Department of Neurology, Children’s Hospital of Shanxi, Taiyuan, Shanxi, People’s Republic of China; 11Pediatric Inpatient Ward, The 2nd Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, People’s Republic of China; 12Department of Infectious Disease, Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai, People’s Republic of China; 13Outpatient Department of Pediatrics, The First Hospital of Jilin University, Changchun, Jilin, People’s Republic of China; 14Department of Pediatric Infection and Gastroenterology, Tongji Hospital, Tongji Medical College of Huazhong University of Science and Technology, Wuhan, Hubei, People’s Republic of China; The CPBMS team: the Children’s Hospital of Zhejiang University School of Medicine; National Clinical Research Center for Child Health; National Children’s Regional Medical Center; Children’s Hospital of Chongqing Medical University; Beijing Children’s Hospital; Capital Medical University; National Center for Children’s Health; Research Unit of Critical Infection in Children; Chinese Academy of Medical Sciences, 2019RU016; Hunan Children’s Hospital; the Children’s Hospital of Fudan University; Anhui Province Children’s Hospital; Children’s Hospital of Nanjing Medical University; West China Second University Hospital; Sichuan University/ West China Women’s and Children’s Hospital; Shanxi Children’s Hospital; the 2nd Affiliated Hospital and Yuying Children’s Hospital of Wenzhou Medical University; Xinhua Hospital Affiliated to Shanghai Jiao Tong University School of Medicine; the First Hospital of Jilin University; the Affiliated Children’s Hospital of Xi’an Jiaotong University; Xi’an Central Hospital; the First Affiliated Hospital of Zhengzhou University; the First Affiliated Hospital of Guangxi Medical University; Jiangxi Provincial Children’s Hospital; Children’s Hospital of Soochow University; Kaifeng Children’s Hospital; Children’s Hospital Affiliated to Shandong University; Hainan Women and Children’s medical center; Women and Children’s Hospital; School of Medicine; Xiamen University; Shanghai Children’s Medical Center; National Children’s Medical Center; Shanghai Jiaotong University School of Medicine; the First Hospital of Lanzhou University; the Second Hospital of Hebei Medical University; the Women’s and Children’s Hospital of Qinghai Province; Taiyuan Maternal and Child Health Care Hospital; the First People’s Hospital of Zunyi; Inner Mongolia People’s Hospital; the Affiliated Hospital of Qingdao University; Hangzhou Children’s Hospital; Beijing Ditan Hospital; Capital Medical University; Tongji Hospital; Tongji Medical College of Huazhong University of Science and TechnologyCorrespondence: Yinghu Chen, Department of Infectious Disease, Children’s Hospital, Zhejiang University School of Medicine, National Clinical Research Center for Child Health, National Children’s Regional Medical Center, 3333 Binsheng Road, Hangzhou, Zhejiang, 310052, People’s Republic of China, Tel +86 13857154891, Email cyh18@zju.edu.cnObjective: To explore the epidemiological and pathogenic characteristics of children with community-acquired bacterial meningitis.Methods: A multicenter, retrospective study was conducted among CABM patients under 15 years old from 33 hospitals in China from 2019 to 2020. The medical record, laboratory, and microbiological data were collected and analyzed.Results: A total of 1610 children with CABM were identified and presented at a median onset age of 45 days of whom 955 (59.3%) were males. CABM occurred mostly in infants < 1 year of age (84.0%, 1352/1610). In etiology-confirmed cases, the pathogens were isolated from CSF culture in 515 (32.0%), 400 (24.8%) in blood culture, and 186 (11.6%) both in CSF and blood culture. In total, 126 pathogens were identified through CSF mNGS in 330 CABM cases; 21 S. pneumoniae isolates were detected in 83 CABM cases by antigen detection method. Major pathogens were E. coli (195, 24.7%), GBS (170, 21.5%), and S. pneumoniae (157, 19.9%). GBS (29.3%, 22/75) was the first pathogen of CABM in neonates aged 0– 6 days old, while E. coli (44.7%, 76/170) in 7 to 28 days of age; S. pneumoniae (96.2%, 151/157) was the most common pathogen in > 3 months old cases. About 9.7% (19/195) strains of E. coli produced ultra‑broad‑spectrum β‑lactamases. The common intracranial imaging complications were subdural effusion and (or) empyema in 349 (21.7%), hydrocephalus in 233 (14.5%), and cerebral abscess in 178 (11.1%). A total of 389 (24.2%) cases were completely cured and 1088 (67.6%) cases improved. Among 166 patients (10.3%) with adverse outcomes, 32 cases (2.0%) died, and 37 cases (2.3%) relapsed.Conclusion: The onset age of CABM in children is usually within 1 year of age, especially < 3 months. The primary pathogens in infants less than 3 months old are E. coli and GBS, and the dominant pathogen in children older than 3 months old is S. pneumoniae. Subdural effusion and (or) empyema and hydrocephalus are common complications. CABM should not be excluded even if CSF leukocyte counts are within normal range. Due to the low detection rate of pathogens in children with CABM, standardized CSF bacteriological examination should be paid more attention to increase the pathogen detection rate. Non‑culture CSF detection methods may facilitate pathogenic diagnosis.Keywords: meningitis, bacterial, pathogen, childhood, diagnosis, outcome

Ling Yang &ast;, Qi Shen &ast;, Chao Hu, Ying Wang, Xiaohong Zhu, Shiqing Shu, Zhu Luo Clinical Trial Center, National Medical Products Administration Key Laboratory for Clinical Research and Evaluation of Innovative Drugs, West China Hospital Sichuan University, Chengdu, People’s Republic of China&ast;These authors contributed equally to this workCorrespondence: Zhu Luo, Clinical Trial Center, National Medical Products Administration Key Laboratory for Clinical Research and Evaluation of Innovative Drugs, West China Hospital Sichuan University, No. 37 Guoxue Lane, Chengdu, Sichuan, 610044, People’s Republic of China, Tel +86 28 85422707, Email luozhu720@163.comBackground: Imrecoxib is a novel and moderately selective cyclooxygenase-2 inhibitor with properties of anti-inflammation and alleviating pain, which is widely applied in osteoarthritis patients. The pharmacokinetic data supporting imrecoxib’s rational use in elderly population are not available.Purpose: The study aims to investigate the pharmacokinetics of imrecoxib and its main metabolites and explore the safety of imrecoxib in elderly healthy subjects.Methods: A total of 19 healthy subjects including 10 non-elderly and 9 elderly subjects received single dose of 100 mg imrecoxib under fasting condition. Pharmacokinetics, safety and tolerability profiles were assessed.Results: After oral administration of single dose of 100 mg imrecoxib, it was absorbed into plasma with median time to reach peak concentration (Tmax) around 2 hours. The concentration–time curves of imrecoxib (M0) showed higher interindividual variability in elderly subjects compared with non-elderly subjects. Peak concentration (Cmax) of M0, its hydroxyl metabolite M1 and carboxylated metabolite M2 in plasma increased by 39%, 21% and 17%, and area under concentration–time curve from time 0 to time t (AUC0-t) of M0, M1 and M2 in plasma increased by 34%, 13% and 27%, respectively, in elderly subjects compared with non-elderly subjects. The 90% CIs of geometric mean ratios of Cmax, AUC0-t and AUC0-∞ of M0, M1 and M2 between the two groups were not located within 80– 125%, indicating Cmax, AUC0-t and AUC0-∞ were not completely equivalent between non-elderly and elderly healthy subjects. However, comparison of pharmacokinetic data of M0, M1 and M2 between the two groups showed no significant difference (P> 0.05). Imrecoxib was well tolerated in both non-elderly and elderly healthy subjects, especially with favorable gastrointestinal and cardiovascular safety profiles.Conclusion: Pharmacokinetic and safety profiles of imrecoxib in elderly healthy subjects indicated that no dose adjustment should be required for elderly population.Keywords: cyclooxygenase-2 inhibitor, imrecoxib, pharmacokinetics, safety, elderly

Yan Meng,1,2 Sheng Wang,1 Wei Zhang,1 Chunlin Xie,1 Xiaoqing Chai,1 Shuhua Shu,1 Yu Zong1,2 1Department of Anesthesiology, The First Affiliated Hospital of University of Science and Technology of China (USTC)/Anhui Provincial Hospital, Hefei, People’s Republic of China; 2Core Facility Center, The First Affiliated Hospital of USTC/Anhui Provincial Hospital, Hefei, People’s Republic of ChinaCorrespondence: Yu Zong, Department of Anesthesiology, The First Affiliated Hospital of USTC, 17 Lujiang Road, Hefei, Anhui, People’s Republic of China, Email zongyu3540@163.comObjective: Continuous interscalene brachial plexus block (cIBPB) is an effective perioperative analgesic therapy for shoulder arthroscopic surgery (SAS) patients. This trial aimed to compare the effect of different cIBPB infusion methods on postoperative analgesia and respiratory function in patients undergoing SAS.Methods: After SAS, 88 patients were randomly assigned to four groups. Through interscalene catheter, all the patients received an initial dose of 10 mL 0.2% ropivacaine. The CI group received 0.2% ropivacaine 4 mL/h, and the PIBI1, PIBI2, and PIBI3 groups received intermittent 0.2% ropivacaine boluses at 4 mL/h, 8 mL/2 h, and 12 mL/3 h, respectively. The patients could also use a patient-controlled analgesia (PCA) pump to self-inject a tramadol bolus each time he/she felt pain. The primary outcome was the cumulative tramadol consumption over the 48 h after surgery. Secondary outcome measures included PCA frequency, pain (visual analogue scale, VAS) score, patient satisfaction, diaphragmatic excursion, pulmonary function, and adverse events.Results: The cumulative tramadol consumption and PCA frequency over the 48 h after surgery in groups PIBI2 and PIBI3 were lower than in both the CI and PIBI1 groups (p< 0.001). The VAS scores (at rest and on movement) in groups PIBI2 and PIBI3 were lower than those in the CI and PIBI1 groups at 8 and 12 h after surgery (all p< 0.001). Patient satisfaction scores were significantly higher in the PIBI2 group than in the other three groups (all p< 0.001). Diaphragmatic excursion was significantly decreased in the PIBI3 group compared to the other three groups (p< 0.05). The incidence of adverse events over the 48 h after surgery was significantly higher in the PIBI3 group compared to the other three groups (p< 0.001).Conclusion: Programmed intermittent bolus infusion with 0.2% ropivacaine 8 mL/2 h for cIBPB can achieve lower tramadol consumption, along with better analgesia after surgery, lower reduction in diaphragmatic excursion, lower incidence of adverse events, and higher patient satisfaction.Keywords: continuous interscalene brachial plexus block, programmed intermittent bolus infusion, shoulder arthroscopic surgery, perioperative analgesic therapy, respiratory function

Jianchang Qian,1,* Sihui Yin,1,* Lin Ye,1 Zhe Wang,2 Sheng Shu,1,3 Zhenxin Mou,1,3 Mingjiang Xu,1 Nipon Chattipakorn,4 Zhiguo Liu,1 Guang Liang1,3,5 1Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, People’s Republic of China; 2Department of Pharmacy, The Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang, People’s Republic of China; 3School of Pharmacy, Hangzhou Medical College, Hangzhou, Zhejiang, People’s Republic of China; 4Cardiac Electrophysiology Research and Training Center, Faculty of Medicine, Chiang Mai University, Chiang Mai, Thailand; 5Wenzhou Institute, University of Chinese Academy of Sciences, Wenzhou, Zhejiang, People’s Republic of China*These authors contributed equally to this workCorrespondence: Guang Liang; Zhiguo LiuChemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, 325035, People’s Republic of ChinaTel/Fax +86-577-86699396Email wzmcliangguang@163.com; lzgcnu@163.comAim: Elevated inflammatory signaling has been shown to play an important role in diabetic kidney disease (DKD). We previously developed a new anti-inflammatory compound LG4. In the present study, we have tested the hypothesis that LG4 could prevent DKD by suppressing inflammation and identified the underlying mechanism.Methods: Streptozotocin-induced type 1 diabetic mice were used to develop DKD and evaluate the effects of LG4 against DKD. To identify the potential targets of LG4, biotin-linked LG4 was synthesized and subjected to proteome microarray screening. The cellular mechanism of LG4 was investigated in HG-challenged SV40MES13 cells.Results: Although LG4 treatment had no effect on the body weight and blood glucose levels, it remarkably reversed the hyperglycemia-induced pathological changes and fibrosis in the kidneys of T1DM mice. Importantly, hyperglycemia-induced renal inflammation evidenced by NF-κB activation and TNFα and IL-6 overexpression was greatly ameliorated with LG4 treatment. Proteosome microarray screening revealed that JNK and ERK were the direct binding proteins of LG4. LG4 significantly reduced HG-induced JNK and ERK phosphorylation and subsequent NF-κB activation in vivo and in vitro. In addition, LG4 did not show further anti-inflammatory effect in HG-challenged mesangial cells with the presence of JNK or ERK inhibitor.Conclusion: LG4 showed renoprotective activity through inhibiting ERK/JNK-mediated inflammation in diabetic mice, indicating that LG4 may be a therapeutic agent for DKD.Keywords: indole-2-carboxamide derivative, diabetic kidney disease, inflammation, MAPK, NF-κB

Xin-lu Yang,1,* Hai Gu,1,* Ji-cheng Hu,1 Sheng Wang,1 Xin Wei,1 Shu-hua Shu,1 Wei-de Zhou,1 Chun-rong Tao,2 Di Wang,1 Xiao-qing Chai1 1Department of Anesthesiology, Pain Clinic, First Affiliated Hospital, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, People’s Republic of China; 2Department of Neurology, First Affiliated Hospital, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, People’s Republic of China*These authors contributed equally to this workCorrespondence: Di Wang; Xiao-qing ChaiDepartment of Anesthesiology, First Affiliated Hospital, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei 230001, People’s Republic of ChinaTel/Fax +86-551-62283963; +86-551-62283912Email di.wang@ustc.edu.cn; xiaoqingchai@ustc.edu.cnPurpose: This randomized, double-blind study evaluated the effectiveness and limitations of continuous serratus anterior plane block (cSAPB) by comparing the effect of cSAPB to patient-controlled intravenous analgesia (PCIA) on postoperative acute pain after thoracoscopic surgery in adults.Patients and Methods: Sixty-six patients who underwent elective video-assisted thoracoscopic surgery (VATS) were randomly allocated to cSAPB or PCIA groups (n=33 per group) after surgery. For the cSAPB group, patients were treated by an initial does of 20 mL ropivacaine (0.375%), followed by continuous infusion at a rate of 5 mL/h of ropivacaine (0.2%) and a patient-controlled bolus of 5 mL ropivacaine (0.2%). PCIA started with an initial does of 0.03 μg/kg sufentanil, followed by a basal infusion of 0.03 μg/kg/h sufentanil and a patient-controlled bolus of 0.03 μg/kg sufentanil. Visual analog scale (VAS) and other items were examined postoperatively. The area under the curve of VAS-time (AUCVAS-time) at rest and on coughing in the first 24 hours postoperatively were primary outcomes.Results: At the first 24 hours postoperatively, patients in the cSAPB group exhibited a smaller AUCVAS-time at rest (44.0± 17.1 vs 68.9± 11.8 cm·h, P< 0.001) and AUCVAS-time on coughing (67.1± 8.8 vs 78.0± 12.5 cm·h, P< 0.001) compared with those in the PCIA group. The differences in means of VAS score at rest were more than 1.0 cm between the two groups, however, on coughing they were less than 1.0 cm at each observation point. Additionally, patients in the cSAPB group had a longer time to first patient-controlled bolus (15.8± 7.6 vs 10.6± 8.6 hours, P=0.011). Furthermore, a higher rank of satisfaction was recorded with patients in the cSAPB group.Conclusion: cSAPB using PCA devices might be superior to traditional intravenous continuous analgesia, particularly with an advantage of pain relief at rest following VATS operation. Meanwhile, cSAPB lacks a satisfactory analgesic effect on cough.Keywords: patient-controlled regional analgesia, continuous regional analgesia, continuous peripheral nerve blocks, postoperative acute pain, video-assisted thoracoscopic surgery

Hui Fei,1,* Xuefeng Jiang,2,* Tian Li,1 Ying Pan,2 Hongling Guo,1 Xiaoyu Xu,1 Shanrong Shu2 1Department of Gynecology and Obstetrics, The Seventh Affiliated Hospital of Sun Yat-Sen University, Shenzhen 518107, People’s Republic of China; 2Department of Gynecology and Obstetrics, The First Affiliated Hospital of JiNan University, Guangzhou 510630, People’s Republic of China*These authors contributed equally to this workCorrespondence: Shanrong ShuDepartment of Gynecology and Obstetrics, The First Affiliated Hospital of JiNan University, 613 Huangpu Road West, Guangzhou 510630, People’s Republic of ChinaTel +86-20-38688769Fax +86-20-38688769Email butterfly1705@126.comBackground/Aim: Cesarean scar pregnancy is a long term complication of cesarean section. There is a lot of controversy about the best treatment methods. We retrospectively summarized the clinical characteristics of patients with cesarean scar pregnancy and explored the advantages and disadvantages of fertility-preservation treatment method.Methods: From January 2008 to September 2017, a total of 204 cases of cesarean scar pregnancy were retrospectively reviewed. 145 patients underwent transvaginal clearance, 33 patients underwent endoscopic surgery, and 26 patients underwent uterine artery embolism. The clinical characteristics, diagnosis, various treatment methods, and clinical outcomes were analyzed.Results: There were no significant differences among the three groups in terms of patient age, number of previous cesarean sections, serum human chorionic gonadotropin, and clinical symptoms. The difference in mean gestational sac diameter (23.5±2.1mm vs 31.3±2.4mm vs 30.8± 1.9mm), surgical time (31.4±2.5min vs. 45.8±2.2min vs. 51.4±1.9min), blood loss (53.3± 5.5mL vs. 105.2±3.2mL vs. 75.6 ±3.5mL), blood transfusion (1/145 case vs.3/33 case vs. 0/26 case), discomfort (1/145 case vs.9/33 case vs. 16/26 case), hospital stay (6.1±1.1 day vs. 7.4±0.9 day vs.18.6±1.5 day), fever duration (1.0±0.5 day vs. 2.1±2.8 day vs. 5.7±3.5 day), and hospital expense (¥ 7825.9±234.9 vs. ¥ 10248.3± 312.9 vs. ¥ 18774.9±243.6) in transvaginal pregnancy tissue clearance, endoscopic surgery, and uterine artery embolism groups were significantly different.Conclusion: Transvaginal clearance is an effective and relatively safe treatment option for patients with cesarean scar pregnancy.Keywords: cesarean scar pregnancy, transvaginal clearance, endoscopy, uterine artery embolism

Xiaohong Xu,1 Vinothkumar Rajamanickam,1 Sheng Shu,1 Zhoudi Liu,1 Tao Yan,1 Jinxin He,1 Zhiguo Liu,1 Guilong Guo,2 Guang Liang,1 Yi Wang1 1Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, People’s Republic of China; 2Department of Surgical Oncology, The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, Zhejiang 325000, People’s Republic of ChinaCorrespondence: Yi WangChemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang 325035, People’s Republic of ChinaTel/Fax +8657785773060Email yi.wang1122@wmu.edu.cnBackground: Triple-negative breast cancer (TNBC) is an aggressive subtype of breast cancer which is associated with poor patient outcome and lack of targeted therapy. Our laboratory has synthesized a series of indole-2-carboxamide derivatives. Among this series, compound LG25 showed a favorable pharmacological profile against sepsis and inflammatory diseases. In the present study, we investigated the chemotherapeutic potential of LG25 against TNBC utilizing in vitro and in vivo models.Methods: Changes in cell viability, cell cycle phases and apoptosis were analyzed using MTT, clonogenic assay, FACS and Western blotting assays. The anti-cancer effects of LG25 were further determined in a xenograft mouse model.Results: Our findings reveal that LG25 reduced TNBC viability in a dose-dependent manner. Flow cytometric and Western blot analyses showed that LG25 enhances G2/M cell cycle arrest and induced cell apoptosis. In addition, LG25 treatment significantly inhibited Akt/mTOR phosphorylation and nuclear translocation of nuclear factor-κB (NF-κB). These inhibitory activities of LG25 were confirmed in mice implanted MDA-MB-231 TNBC cells.Conclusion: Our studies provide experimental evidence that indole-2-carboxamide derivative LG25 effectively targeted TNBC in preclinical models by inducing cell cycle arrest and apoptosis, through suppressing Akt/mTOR/NF-κB signaling pathway.Keywords: indole analog, triple-negative breast cancer, cell cycle arrest, apoptosis, Akt, mTOR

Siyang Xiao,1,* Wenxin Zhang,1,* Hongjin Chen,1 Bo Fang,1 Yinda Qiu,2 Xianxin Chen,1 Lingfeng Chen,1 Sheng Shu,1 Yunjie Zhao,1 Zhiguo Liu,1 Guang Liang1 1Chemical Biology Research Center, School of Pharmaceutical Sciences, Wenzhou Medical University, Wenzhou, Zhejiang, China; 2College of Life and Environmental Science, Wenzhou University, Wenzhou, Zhejiang, China *These authors contributed equally to this work Purpose: The purpose of this study was to design and synthesize novel 2-benzylidene-1-indanone derivatives for treatment of acute lung injury. Methods: A series of 39 novel 2-benzylidene-indanone structural derivatives were synthesized and evaluated for anti-inflammatory activity in lipopolysaccharide (LPS)-stimulated murine primary macrophages. Results: Most of the obtained compounds effectively inhibited the LPS-induced expression of IL-6 and TNF-α. The most active compound, 8f, was found to significantly reduce LPS-induced pulmonary inflammation, as reflected by reductions in the concentration of total protein, inflammatory cell count, as well as the lung wet/dry ratio in bronchoalveolar lavage (BAL) fluid. Furthermore, 8f effectively inhibited mRNA expression of several inflammatory cytokines after LPS challenge in vitro and in vivo. Administration of 8f also blocked LPS-induced activation of the proinflammatory NF-κB/MAPK signaling pathway. Conclusion: The simple synthetic preparation and biological properties of these derivatives make these 2-benzylidene-indanone scaffolds promising new entities for the development of anti-inflammatory therapeutics for the treatment of acute lung injury. Keywords: indanone, acute lung injury, drug design, anti-inflammation, synthesis

PSZ2 G091.83+26.11 is a massive galaxy cluster with M500 = 7:43 × 1014M⊙ at z = 0:822. This object exhibits a complex morphology with a clear bimodality observed in X-rays. However, it was detected and analysed in the Planck sample as a single, spherical cluster following a universal profile [1]. This model can lead to miscalculations of thermodynamical quantities, like the pressure profile. As future multiwavelength cluster experiments will detect more and more objects at higher redshifts (where we expect the fraction of merging objects to be higher), it is crucial to quantify this systematic effect. In this work, we use high-resolution observations of PSZ2 G091.83+26.11 by the NIKA2 camera to integrate the morphological characteristics of the cluster in our modelling. This is achieved by fitting a two-halo model to the SZ image and then by reconstruction of the resulting projected pressure profile. We then compare these results with the spherical assumption.

High-mass stars (m* ≳ 8 M⊙) play a crucial role in the evolution of galaxies, and so it is imperative that we understand how they are formed. We have used the New IRAM KIDs Array 2 (NIKA2) camera on the Institut de Radio Astronomie Millimétrique (IRAM) 30-m telescope to conduct high-sensitivity continuum mapping of ~ 2 deg2 of the Galactic plane (GP) as part of the Galactic Star Formation with NIKA2 (GASTON) large program. We have identified a total of 1467 clumps within our deep 1.15 mm continuum maps and, by using overlapping continuum, molecular line, and maser parallax data, we have determined their distances and physical properties. By placing them upon an approximate evolutionary sequence based upon 8 μm Spitzer imaging, we find evidence that the most massive dense clumps accrete material from their surrounding environment during their early evolution, before dispersing as star formation advances, supporting clump-fed models of high-mass star formation.

We present preliminary results from an on-going program that aims at mapping the intracluster medium (ICM) temperature of high redshift galaxy clusters from the MaDCoWS sample using a joint analysis of shallow X-ray data obtained by Chandra and high angular resolution Sunyaev-Zel’dovich (SZ) observations realized with the NIKA2 and MUSTANG-2 cameras. We also present preliminary results from an on-going Open Time program within the NIKA2 collaboration that aims at mapping the ICM temperature of a galaxy cluster at z = 0.45 from the resolved detection of the relativistic corrections to the SZ spectrum. These studies demonstrate how high angular resolution SZ observations will play a major role in the coming decade to push the investigation of ICM dynamics and non-gravitational processes to high redshift before the next generation X-ray observatories come into play.

The NIKA2 Guaranteed-Time SZ Large Program (LPSZ) is dedicated to the high-angular resolution SZ mapping of a representative sample of 45 SZ-selected galaxy clusters drawn from the catalogues of the Planck satellite, or of the Atacama Cosmology Telescope. The LPSZ sample spans a mass range from 3 to 11 × 1014M⊙ and a redshift range from 0:5 to 0:9, extending to higher redshift and lower mass the previous samples dedicated to the cluster mass calibration and universal properties estimation. The main goals of the LPSZ are the measurement of the average radial profile of the ICM pressure up to R500 by combining NIKA2 with Planck or ACT data, and the estimation of the scaling law between the SZ observable and the mass using NIKA2, XMM-Newton and Planck/ACT data. Furthermore, combining LPSZ data with existing or forthcoming public data in lensing, optical/NIR or radio domains, we will build a consistent picture of the cluster physics and further gain knowledge on the mass estimate as a function of the cluster morphology and dynamical state. We give an overview of the LPSZ, present recent results and discuss the future implication for cosmology with galaxy clusters.

New observations of the edge-on galaxy NGC 891, at 1.15 and 2mm obtained with the IRAM 30-m telescope and the NIKA2 camera, within the framework of the IMEGIN (Interpreting the Millimetre Emission of Galaxies with IRAM and NIKA2) Large Program, are presented in this work. By using multiwavelength maps (from the mid-IR to the cm wavelengths) we perform SED fitting in order to extract the physical properties of the galaxy on both global and local (~kpc) scales. For the interpretation of the observations we make use of a state-of-the-art SED fitting code, HerBIE (HiERarchical Bayesian Inference for dust Emission). The observations indicate a galaxy morphology, at mm wavelengths, similar to that of the cold dust emission traced by submm observations and to that of the molecular gas. The contribution of the radio emission at the NIKA2 bands is very small (negligible at 1.15 mm and ~ 10% at 2 mm) while it dominates the total energy budget at longer wavelengths (beyond 5 mm). On local scales, the distribution of the free-free emission resembles that of the dust thermal emission while the distribution of the synchrotron emission shows a deficiency along the major axis of the disc of the galaxy.

Interstellar dust plays an important role in the formation of molecular gas and the heating and cooling of the interstellar medium. The spatial distribution of the mm-wavelength dust emission from galaxies is largely unexplored. The NIKA2 Guaranteed Time Project IMEGIN (Interpreting the Millimeter Emission of Galaxies with IRAM and NIKA2) has recently mapped the mm emission in the grand design spiral galaxy NGC6946. By subtracting the contributions from the free-free, synchrotron, and CO line emission, we map the distribution of the pure dust emission at 1:15mm and 2mm. Separating the arm/interarm regions, we find a dominant 2mm emission from interarms indicating the significant role of the general interstellar radiation field in heating the cold dust. Finally, we present maps of the dust mass, temperature, and emissivity index using the Bayesian MCMC modeling of the spectral energy distribution in NGC6946.

The quest for primordial gravitational waves enclosed in the Cosmic Microwave Background (CMB) polarization B-modes signal motivates the development of a new generation of high sensitive experiments (e.g. CMBS4, LiteBIRD), thus allowing to probe the inflationary epoch in the early Universe. However, this will be only possible by ensuring a high control of the instrumental systematic effects and an accurate absolute calibration of the polarization angle. The Crab nebula is known to be a polarization calibrator on the sky for CMB experiments. Already used for the Planck satellite it exhibits a high polarized signal at microwave wavelengths. In this work we present Crab polarization observations obtained, in the 260 GHz frequency band, with the NIKA2 instrument. Furthermore, we discuss the accuracy needed on such a measurement to improve the constraints on the absolute angle calibration for CMB experiments.

Starting from the clusters included in the NIKA sample and in the NIKA2 Sunyaev-Zel’dovich Large Program (LPSZ) we have selected a sample of six common objects with the Cluster Lensing And Supernova survey with Hubble (CLASH) lensing data. For the LPSZ clusters we have at our disposal both high-angular resolution observations of the thermal SZ with NIKA and NIKA2 and X-ray observations with XMM-Newton from which hydrostatic mass estimates can be derived. In addition, the CLASH dataset includes lensing convergence maps that can be converted into lensing estimates of the total mass of the cluster. One-dimensional mass profiles are used to derive integrated mass estimates accounting for systematic effects (data processing, modeling, etc.). Two-dimensional analysis of the maps can reveal substructures in the cluster and, therefore, inform us about the dynamical state of each system. Moreover, we are able to study the hydrostatic mass to lensing mass bias, across different morphology and a range of redshift clusters to give more insight on the hydrostatic mass bias. The analysis presented in this proceeding follows the study discussed in [20].

As the possible progenitors of passive galaxies at z=2-3, dusty starforming galaxies (DSFGs) at z>4 provide a unique perspective to study the formation, assembly, and early quenching of massive galaxies in the early Universe. The extreme obscuration in optical-IR makes (sub)mm spectral scans the most universal and unbiased way to confirm/exclude the high-z nature of candidate dusty star-forming galaxies. We present here the status of the NIKA2 Cosmological Legacy Survey (N2CLS), which is the deepest wide-area singledish survey in the millimeter searching for high-z DSFGs. We also introduce a joint-analysis method to effciently search for the spectroscopic redshift of high-z DSFGs with noisy spectra and photometric data and present its success in identifying the redshift of DSFGs found in NIKA2 science verification data.

Dust polarization observations are a powerful, practical tool to probe the geometry (and to some extent, the strength) of magnetic fields in starforming regions. In particular, Planck polarization data have revealed the importance of magnetic fields on large scales in molecular clouds. However, due to insufficient resolution, Planck observations are unable to constrain the B-field geometry on prestellar and protostellar scales. The high angular resolution of 11.7 arcsec provided by NIKA2-Pol 1.15 mm polarimetric imaging, corresponding to 0.02 pc at the distance of the Orion molecular cloud (OMC), makes it possible to advance our understanding of the B-field morphology in star-forming filaments and dense cores (IRAM 30m large program B-FUN). The commissioning of the NIKA2-Pol instrument has led to several challenging issues, in particular, the instrumental polarization or intensity-to-polarization “leakage” effect. In the present paper, we illustrate how this effect can be corrected for, leading to reliable exploitable data in a structured, extended source such as OMC-1. We present a statistical comparison between NIKA2-Pol and SCUBA2-Pol2 results in the OMC-1 region. We also present tentative evidence of local pinching of the B-field lines near Orion-KL, in the form of a new small-scale hourglass pattern, in addition to the larger-scale hourglass already seen by other instruments such as Pol2.

We present a multi-probe analysis of the well-known galaxy cluster CL J1226.9+3332 as a proof of concept for multi-wavelength studies within the framework of the NIKA2 Sunyaev-Zel’dovich Large Program (LPSZ). CL J1226.9+3332 is a massive and high redshift (z = 0.888) cluster that has already been observed at several wavelengths. A joint analysis of the thermal SZ (tSZ) effect at millimeter wavelength with the NIKA2 camera and in X-ray with the XMM-Newton satellite permits the reconstruction of the cluster’s thermodynamical properties and mass assuming hydrostatic equilibrium. We test the robustness of our mass estimates against different definitions of the data analysis transfer function. Using convergence maps reconstructed from the data of the CLASH program we obtain estimates of the lensing mass, which we compare to the estimated hydrostatic mass. This allows us to measure the hydrostatic-to-lensing mass bias and the associated systematic effects related to the NIKA2 measurement. We obtain M500HSE = (7:65 ± 1:03) × 1014M⊙ and M500lens = (7:35 ± 0:65) × 1014M⊙, which implies a HSE-to-lensing bias consistent with 0 within 20%.

Using the NIKA2 dual band millimeter camera installed on the IRAM30m telescope, we have mapped a relatively large field (~ 70 arcmin2) in the direction of the star GJ526 to investigate the nature of the sources found with the MAMBO camera at 1.2 mm ten years earlier. We have found that they must be dust-obscured galaxies (SMGs) in the background beyond the star. The new NIKA2 map at 1.15 mm reveals additional sources and, in fact, an overdensity of SMGs predominantly distributed along a filament-like structure in projection on the sky across the whole observed field. We speculate this might be a cosmic filament at high redshift as revealed in cosmological hydrodynamical simulations. Measurement of spectroscopic redshifts of the SMGs in the candidate filament is required now for a definitive confirmation of the nature of the structure.

In the past decade filaments have been recognised as a major structural element of the interstellar medium, the densest of these filaments hosting the formation of most stars. In some star-forming molecular clouds converging networks of filaments, also known as hub filament systems, can be found. These hubs are believed to be preferentially associated to massive star formation. As of today, there are no metrics that allow the systematic quantification of a filament network convergence. Here, we used the IRAM 30m NIKA2 observations of the Galactic plane from the GASTON large programme to systematically identify filaments and produce a filament convergence parameter map. We use such a map to show that: i. hub filaments represent a small fraction of the global filament population; ii. hubs host, in proportion, more massive and more luminous compact sources that non-hubs; iii. hub-hosting clumps are more evolved that non-hubs; iv. no discontinuities are observed in the properties of compact sources as a function of convergence parameter. We propose that the rapid global collapse of clumps is responsible for (re)organising filament networks into hubs and, in parallel, enhancing the mass growth of compact sources.

We present here the polarization system of the NIKA camera and give a summary of the main results obtained and performed studies on Orion and the Crab nebula. The polarization system was equipped with a room temperature continuously rotating multi-mesh half wave plate and a grid polarizer facing the NIKA cryostat window. NIKA even though less sensitive than NIKA2 had polarization capability in both 1 and 2 millimiter bands. NIKA polarization observations demonstrated the ability of such a technology in detecting the polarization of different targets, compact and extended sources like the Crab nebula and Orion Molecular Cloud region OMC-1. These measurements together with the developed techniques to deal with systematics, opened the way to the current observations of NIKA2 in polarization that will provide important advances in the studies of galactic and extra-galactic emission and magnetic fields.

The main limiting factor of cosmological analyses based on thermal Sunyaev-Zel’dovich (SZ) cluster statistics comes from the bias and systematic uncertainties that affect the estimates of the mass of galaxy clusters. High-angular resolution SZ observations at high redshift are needed to study a potential redshift or morphology dependence of both the mean pressure profile and of the mass-observable scaling relation used in SZ cosmological analyses. The NIKA2 camera is a new generation continuum instrument installed at the IRAM 30-m telescope. With a large field of view, a high angular resolution and a high-sensitivity, the NIKA2 camera has unique SZ mapping capabilities. In this paper, we present the NIKA2 SZ large program, aiming at observing a large sample of clusters at redshifts between 0.5 and 0.9, and the characterization of the first cluster oberved with NIKA2.

The NIKA2 polarization channel at 260 GHz (1.15 mm) has been proposed primarily to observe galactic star-forming regions and probe the critical scales between 0.01-0.05 pc at which magnetic field lines may channel the matter of interstellar filaments into growing dense cores. The NIKA2 polarime-ter consists of a room temperature continuously rotating multi-mesh HWP and a cold polarizer that separates the two orthogonal polarizations onto two 260 GHz KIDs arrays. We describe in this paper the preliminary results obtained during the most recent commissioning campaign performed in December 2018. We concentrate here on the analysis of the extended sources, while the observation of compact sources is presented in a companion paper [12]. We present preliminary NIKA2 polarization maps of the Crab nebula. We find that the integrated polarization intensity flux measured by NIKA2 is consistent with expectations. In terms of polarization angle, we are still limited by systematic uncertainties that will be further investigated in the forthcoming commissioning campaigns.

Clarifying the role of magnetic fields in the star formation process is crucial. Observations have already shown that magnetic fields play an important role in the early stages of star formation. The high spatial resolution (∼0.01 to 0.05 pc) provided by NIKA2-Pol 1.2 mm imaging polarimetry of nearby clouds will help us clarify the geometry of the B-field within dense cores and molecular filaments as part of the IRAM 30m large program B-FUN. There are numerous challenging issues in the validation of NIKA2-Pol such as the calibration of instrumental polarization. The commissioning phase of NIKA2-Pol is underway and is helping us characterize the intensity-to-polarization “leakage” pattern of the instrument. We present a preliminary analysis of the leakage pattern and its dependence with elevation. We also present the current leakage correction made possible by the NIKA2 pipeline in polarization mode based on the NIKA2-Pol commissioning data taken in December 2018. Based on reduced Stokes I, Q, U data we find that the leakage pattern of NIKA2-Pol depends on elevation and is sensitive to the focus of the telescope.

Understanding where and when the mass of stars is determined is one of the fundamental, mostly unsolved, questions in astronomy. Here, we present the first results of GASTON, the Galactic Star Formation with NIKA2 large programme on the IRAM 30m telescope, that aims to identify new populations of low-brightness sources to tackle the question of stellar mass determination across all masses. In this paper, we focus on the high-mass star formation part of the project, for which we map a ~ 2 deg2 region of the Galactic plane around l = 24° in both 1.2 mm and 2.0 mm continuum. Half-way through the project, we reach a sensitivity of 3.7 mJy/beam at 1.2mm. Even though larger than our target sensitivity of 2 mJy, the current sensitivity already allows the identification of a new population of cold, compact sources that remained undetected in any (sub-)mm Galactic plane survey so far. In fact, about 25% of the ~ 1600 compact sources identified in the 1.2mm GASTON image are new detections. We present a preliminary analysis of the physical properties of the GASTON sources as a function of their evolutionary stage, arguing for a potential evolution of the mass distribution of these sources with time.

High-resolution mapping of the hot gas in galaxy clusters is a key tool for cluster-based cosmological analyses. Taking advantage of the NIKA2 millimeter camera operated at the IRAM 30-m telescope, the NIKA2 SZ Large Program seeks to get a high-resolution follow-up of 45 galaxy clusters covering a wide mass range at high redshift in order to re-calibrate some of the tools needed for the cosmological exploitation of SZ surveys. We present the second cluster analysis of this program, targeting one of the faintest sources of the sample in order to tackle the difficulties in data reduction for such faint, low-SNR clusters. In this study, the main challenge is the precise estimation of the contamination by sub-millimetric point sources, which greatly affects the tSZ map of the cluster. We account for this contamination by performing a joint fit of the SZ signal and of the flux density of the compact sources. A prior knowledge of these fluxes is given by the adjustment of the SED of each source using data from both NIKA2 and the Herschel satellite. The first results are very promising and demonstrate the possibility to estimate thermodynamic properties with NIKA2, even in a compact cluster heavily contaminated by point sources.

Clusters of galaxies, the largest bound objects in the Universe, constitute a cosmological probe of choice, which is sensitive to both dark matter and dark energy. Within this framework, the Sunyaev-Zel’dovich (SZ) effect has opened a new window for the detection of clusters of galaxies and for the characterization of their physical properties such as mass, pressure and temperature. NIKA, a KID-based dual band camera installed at the IRAM 30-m telescope, was particularly well adapted in terms of frequency, angular resolution, field-of-view and sensitivity, for the mapping of the thermal and kinetic SZ effect in high-redshift clusters. In this paper, we present the NIKA cluster sample and a review of the main results obtained via the measurement of the SZ effect on those clusters: reconstruction of the cluster radial pressure profile, mass, temperature and velocity.

The new NIKA2 camera at the IRAM 30m radiotelescope was used to observe three known debris disks in order to constrain the SED of their dust emission in the millimeter wavelength domain. We have found that the spectral index between the two NIKA2 bands (1mm and 2mm) is consistent with the Rayleigh-Jeans regime (λ-2), unlike the steeper spectra (λ-3) measured in the submillimeter-wavelength domain for two of the three disks - around the stars Vega and HD107146. We provide a succesful proof of concept to model this spectral inversion in using two populations of dust grains, those smaller and those larger than a grain radius a0 of 0.5mm. This is obtained in breaking the slope of the size distribution and the functional form of the absorption coefficient of the standard model. The third disk - around the star HR8799 - does not exhibit this spectral inversion but is also the youngest.

We present the results of the analysis of the very massive cluster MOO J1142+1527 at a redshift z = 1.2 based on high angular resolution NIKA2 Sunyaev-Zel’dovich (SZ) and Chandra X-ray data. This multi-wavelength analysis enables us to estimate the shape of the temperature profile with unprecedented precision at this redshift and to obtain a map of the gas entropy distribution averaged along the line of sight. The comparison between the cluster morphological properties observed in the NIKA2 and Chandra maps together with the analysis of the entropy map allows us to conclude that MOOJ1142+1527 is an on-going merger hosting a cool-core at the position of the X-ray peak. This work demonstrates how the addition of spatially-resolved SZ observations to low signal-to-noise X-ray data can bring valuable insights on the intracluster medium thermodynamic properties at z > 1.

The steps taken to tailor to NIKA2 observations the Scanamorphos algorithm (initially developed to subtract low-frequency noise from Herschel on-the-fly observations) are described, focussing on the consequences of the different instrument architecture and observation strategy. The method, making the most extensive use of the redundancy built in the multi-scan coverage with large arrays of a given region of the sky, is applicable to extended sources, while the pipeline is so far optimized for compact sources. An example of application is given. A related tool to build consistent broadband SEDs from 60 microns to 2 mm, combining Herschel and NIKA2 data, has also been developed. Its main task is to process the data least affected by low-frequency noise and coverage limitations (i.e. the Herschel data) through the same transfer function as the NIKA2 data, simulating the same scan geometry and applying the same noise and atmospheric signal as extracted from the 1mm and 2mm data.

Shan-rong Shu, Xin Luo, Zhi-xin Wang, Yu-hong Yao Department of Obstetrics and Gynecology, The First Affiliated Hospital of JiNan University, HuangPu Road West, Guangzhou, People’s Republic of China Abstract: Pregnancy in a cesarean scar is the rarest form of an ectopic pregnancy. The treatment for cesarean scar pregnancy mainly includes systemic methotrexate and uterine artery embolization. Here, we reported a case of cesarean scar pregnancy treated by curettage and aspiration guided by laparoscopy. The treatment plan included two phases. Three days after a combination of methotrexate and mifepristone was administered, the gestational sac was removed under laparoscopy, which enabled a successful treatment for the unruptured ectopic pregnancy in a previous cesarean scar and made it possible to preserve the reproductive capability of the patient. Keywords: cesarean scar pregnancy, laparoscopy, curettage and aspiration

Previous studies have compared the immune systems of wild and of laboratory rodents in an effort to determine how laboratory rodents differ from their naturally occurring relatives. This comparison serves as an indicator of what sorts of changes might exist between modern humans living in Western culture compared to our hunter-gatherer ancestors. However, immunological experiments on wild-caught animals are difficult and potentially confounded by increased levels of stress in the captive animals. In this study, the humoral immune responses of laboratory rats in a traditional laboratory environment and in an environment with enriched biodiversity were examined following immunization with a panel of antigens. Biodiversity enrichment included colonization of the laboratory animals with helminths and co-housing the laboratory animals with wild-caught rats. Increased biodiversity did not apparently affect the IgE response to peanut antigens following immunization with those antigens. However, animals housed in the enriched biodiversity setting demonstrated an increased mean humoral response to T-independent and T-dependent antigens and increased levels of "natural" antibodies directed at a xenogeneic protein and at an autologous tissue extract that were not used as immunogens.