BACKGROUND:Hepatic echinococcosis is a severe endemic disease in some underdeveloped rural areas worldwide. Qualified physicians are in short supply in such areas, resulting in low rates of accurate diagnosis of this condition. In this study, we aimed to develop and evaluate an artificial intelligence (AI) system for automated detection and subtyping of hepatic echinococcosis using plain CT images with the goal of providing interpretable assistance to radiologists and clinicians. METHODS:We developed EDAM, an echinococcosis diagnostic AI system, to provide accurate and generalisable CT analysis for distinguishing hepatic echinococcosis from hepatic cysts and normal controls (no liver lesions), as well as subtyping hepatic echinococcosis as alveolar or cystic echinococcosis. EDAM includes a slice-level prediction model for lesion classification and segmentation and a patient-level diagnostic model for patient classification. We collected a plain CT database (n=700: 395 cystic echinococcosis, 122 alveolar echinococcosis, 130 hepatic cysts, and 53 normal controls) for developing EDAM, and two additional independent cohorts (n=156) for external validation of its performance and generalisation ability. We compared the performance of EDAM with 52 experienced radiologists in diagnosing and subtyping hepatic echinococcosis. FINDINGS:EDAM showed reliable performance in patient-level diagnosis on both the internal testing data (overall area under the receiver operating characteristic curve [AUC]: 0·974 [95% CI 0·936-0·994]; accuracy: 0·952 [0·939-0·965] for cystic echinococcosis, 0·981 [0·973-0·989] for alveolar echinococcosis; sensitivity: 0·966 [0·951-0·979] for cystic echinococcosis, 0·944 [0·908-0·970] for alveolar echinococcosis) and the external testing set (overall AUC: 0·953 [95% CI 0·840-0·973]; accuracy: 0·929 [0·915-0·947] for cystic echinococcosis, 0·936 [0·919-0·950] for alveolar echinococcosis; sensitivity: 0·913 [0·879-0·944] for cystic echinococcosis, 0·868 [0·841-0·897] for alveolar echinococcosis). The sensitivity of EDAM was robust across images from different CT manufacturers. EDAM outperformed most of the enrolled radiologists in detecting both alveolar echinococcosis and cystic echinococcosis. INTERPRETATION:EDAM is a clinically applicable AI system that can provide patient-level diagnoses with interpretable results. The accuracy and generalisation ability of EDAM demonstrates its potential for clinical use, especially in underdeveloped areas. FUNDING:Project of Qinghai Provincial Department of Science and Technology of China, National Natural Science Foundation of China, and Tsinghua-Fuzhou Institute of Data Technology Project. TRANSLATION:For the Chinese translation of the abstract see Supplementary Materials section.

Hepatocellular carcinoma（HCC） has an insidious onset, and most patients are in the advanced stage when attending the hospital and thus lose the opportunity for radical surgical resection, which results in the poor prognosis of patients. With the development of clinical treatment, the treatment of advanced HCC has gradually transitioned from the relatively single and limited treatment options in the past to the new model of comprehensive treatment. In recent years, immunotherapy, represented by immune checkpoint inhibitors（ICIs）, has become widely used in clinical practice. At present, a number of clinical studies have been conducted for immunotherapy combined with local and targeted antitumor therapy, and in particular, ICIs combined with targeted therapy have become a research hotspot in the field of HCC treatment. This article reviews the research advances in immunotherapy for the treatment of HCC.

Hepatic alveolar echinococcosis (HAE) is a zoonotic parasitic disease caused by the larvae of Echinococcus multilocularis. Because of its characteristics of diffuse infiltration and growth similar to tumors, the disability rate and mortality rate are high among patients. Although surgery (including hepatectomy, liver transplantation, and autologous liver transplantation) is the first choice for the treatment of hepatic alveolar echinococcosis in clinic, drug treatment still plays an important and irreplaceable role in patients with end-stage echinococcosis, including patients with multiple organ metastasis, patients with inferior vena cava invasion, or patients with surgical contraindications, etc. However, Albendazole is the only recommended clinical drug which could exhibit a parasitostatic rather than a parasitocidal effect. Novel drugs are needed but few investment was made in the field because the rarity of the cases. Drug repurposing might be a solution. In this review, FDA-approved drugs that have a potential curative effect on hepatic alveolar echinococcosis in animal models are summarized. Further, nano drug delivery systems boosting the therapeutic effect on hepatic alveolar echinococcosis are also reviewed. Taken together, these might contribute to the development of novel strategy for advanced hepatic alveolar echinococcosis.

BACKGROUND: Alveolar echinococcosis is an epidemic disease caused by the parasitism of Echinococcus multilocularis (Em) larvae in the intermediate or final host. OBJECTIVE: To identify and analyze B-cell and T-cell (Th1, Th2, and Th17) epitopes of the Em antigen protein thrombospondin 3 (TSP3). METHODS: The amino acid sequence of TSP3 was obtained, and the secondary structural characteristics of TSP3 were predicted using bioinformatics software to further predict its potential T-cell and B-cell epitopes. The spleen lymphocytes of BALB/c mice, which were immunized with the TSP3 protein, were collected for co-culture with B-cell and T-cell antigen small peptides. The B-cell epitopes and T-cell epitope subtypes Th1, Th2, and Th17 were identified as having good immunogenicity. RESULTS: After identification, it was found that the predominant epitopes of B cells existing in TSP3 were T18-33, T45-55, and T110-122. Furthermore, the predominant epitopes of T cells existing in TSP3 were T33-42, T45-55, T80-90, and T110-122 in the T1 subtype, T45-55, T68-77, and T92-104 in the Th2 subtype, and T53-63 and T80-90 in the Th17 subtype. CONCLUSIONS: Six T-cell and eight B-cell dominant epitopes of the TSP3 antigen were revealed; these results may be applied in the development of a dominant epitope vaccine. Keywords Echinococcus multilocularis , bioinformatics , epitopes

BACKGROUND: Alveolar echinococcosis is an epidemic disease caused by the parasitism of Echinococcus multilocularis (Em) larvae in the intermediate or final host. OBJECTIVE: To identify and analyze B-cell and T-cell (Th1, Th2, and Th17) epitopes of the Em antigen protein thrombospondin 3 (TSP3). METHODS: The amino acid sequence of TSP3 was obtained, and the secondary structural characteristics of TSP3 were predicted using bioinformatics software to further predict its potential T-cell and B-cell epitopes. The spleen lymphocytes of BALB/c mice, which were immunized with the TSP3 protein, were collected for co-culture with B-cell and T-cell antigen small peptides. The B-cell epitopes and T-cell epitope subtypes Th1, Th2, and Th17 were identified as having good immunogenicity. RESULTS: After identification, it was found that the predominant epitopes of B cells existing in TSP3 were T18-33, T45-55, and T110-122. Furthermore, the predominant epitopes of T cells existing in TSP3 were T33-42, T45-55, T80-90, and T110-122 in the T1 subtype, T45-55, T68-77, and T92-104 in the Th2 subtype, and T53-63 and T80-90 in the Th17 subtype. CONCLUSIONS: Six T-cell and eight B-cell dominant epitopes of the TSP3 antigen were revealed; these results may be applied in the development of a dominant epitope vaccine.

Echinococcosis is a human-animal parasitic disease caused by Echinococcosis tapeworm larvae in humans. From a global perspective, it is mainly prevalent in the mid-high latitudes of the Northern Hemisphere, and it is a widespread infectious disease. Its form, host and release areas are slightly different. In clinical practice, Echinococcus granulosus (hepatic cystic echinococcosis) is the most common. Its growth mode is swelling growth and its metastasis is more common in implanted metastasis; However, hepatic alveolar echinococcosis (HAE) is rare. It has been reported that HAE can metastasize through the blood or lymph nodes, and its invasive growth pattern is known as "carcinoma". At this time, it may be accompanied by invasion of the portal vein and inferior vena cava(IVC)or metastasis to distant organs outside the liver (such as lung, brain, lymph nodes). Most patients are in the middle or late stages, making treatment complicated. World Health Organization guidelines recommend radical resection of HAE; However, there is no consensus on lymph node dissection. To date, there have been no reports of cases of HAE accompanied by inferior vena cava-para-abdominal aortic suspected lymph node metastasis and infection. This article reports a clinical case of a complex HAE treated by the surgical method of "middle liver resection + abdominal enlarged lymph node resection + inferior vena cava repair", and histological examination was performed to illustrate the differences in microscopic pathology of alveolar echinococcosis invading the liver and lymph nodes at different magnifications. This article reviews the relevant literature on HAE and derives the latest treatment methods for HAE to provide a reference for future clinical cases of similar alveolar echinococcosis and maximize the benefits of patients.

目的 观察七氟烷吸入溶液剂用于高海拔地区藏族肝包虫手术患者的临床疗效及安全性.方法 将89例拟进行肝包虫手术的高海拔地区藏族患者随机分为对照组43例和试验组46例.对照组给予咪达唑仑2 mg+0.15 mg·kg-1顺式阿曲库铵+2 mg·kg-1丙泊酚+舒芬太尼20～30μg进行麻醉诱导,术中给予瑞芬太尼和丙泊酚分别以25～150μg·kg-1·min-1和0.01～0.10μg·kg-1·min-1的速度静脉麻醉维持,并视术中情况追加顺式阿曲库铵.试验组在麻醉诱导时联合吸入30％七氟烷30 min,其余治疗方法同对照组.比较2组患者的谷丙转氨酶(GPT)和谷草转氨酶(GOT)水平,以及药物不良反应的发生情况.结果 在阻断肝门后10 min、恢复肝血流20 min、恢复肝血流1 h和术后12 h时,试验组的GPT分别为(89.82±21.19),(120.01±30.02),(153.21±44.39)和(230.02±56.28)U·L-1,GOT分别为(78.82±19.20),(115.50±33.40),(162.20±50.03)和(204.03±60.38)U·L-1;对照组的GPT分别为(102.29±25.59),(167.28±29.89),(230.11±52.31)和(365.21±66.54)U·L-1,GOT分别为(101.10±23.30),(153.32±35.50),(210.15±49.82)和(290.88±59.82)U·L-1.2组患者相同时间点的上述指标比较,差异均有统计学意义(均P<0.05).试验组术后出现的药物不良反应有咳嗽和恶心呕吐,对照组术后出现的药物不良反应有恶心呕吐、咳嗽和头痛.试验组和对照组的总药物不良反应发生率分别为17.39％和16.28％,差异无统计学意义(P>0.05).结论 七氟烷吸入溶液剂用于高海拔地区藏族患者肝包虫手术患者具有肝保护作用,且不增加药物不良反应的发生率.

Objective:To evaluate the application value of laparoscopic hydatid cyst punture drainage with Trocar for hepatic cystic echinococcosis.Methods:Clinical data of 20 patients with hepatic cystic echinococcosis treated with laparoscopic cyst punture drainage using disposable Trocar in Qinghai University Affiliated Hospital from January 2018 to January 2020 were retrospectively analyzed. The informed consents of all patients were obtained and the local ethical committee approval was received. Among them, 12 patients were male and 8 female, aged from 10 to 68 years, with a median age of 43 years. The hydatid cysts of 7 cases were located in the left lobe and 13 in the right lobe. 16 cases were with single lesion and 4 with multiple lesions. The median diameter of cysts was 10.3(1.0-18.2) cm. Under laparoscope, a 5-mm disposable Trocar was used to puncture the hydatid cysts through abdominal wall, and a common aspirator was inserted to discharge the cyst fluid for decompression. Hypertonic saline (20%) was injected through the Trocar inlet and soaked for 10 min. Different operations were performed on the patients according to different lesions. The operation results were observed.Results:The operations were completed successfully in all the patients without conversion to open surgery. Excision of internal capsule was performed in 9 cases, excision of internal capsule + dissection of partial external capsule in 6 cases, excision of internal capsule + dissection of partial external capsule + suture of bile leakage in 4 cases, multiple cyst resection in 1 case. Bile leakage occurred in 2 cases and residual cavity effusion in 2 cases after operation, and they were cured after drainage and anti-infection treatments. The remaining patients were properly recovered, and the drainage tube was removed after 3-7 d. The length of postoperative hospital stay was 7(3-12) d. During the postoperative follow-up for 3 months to 1 year, no recurrence or implantation metastasis was reported.Conclusions:Laparoscopic hydatid cyst punture drainage with Trocar for hepatic cystic echinococcosis is an effective, safe and convenient method, which can prevent the fluid leakage and reduce the recurrent risk.

Hydatidosis is a zoonotic parasitic disease caused byEchinococcus, which is highly prevalent in pastoral areas. In China, this disease is mainly caused byEchinococcus granulosusandEchinococcus multilocularis. Cystic echinococcosis, which is one of the most common types of echinococcosis, is described as swelling and growth of cystic lesions. Alveolar echinococcosis, which is less common, is invasive. Cases of mixed echinococcosis infection accompanied by extrahepatic organ metastasis are extremely rare. Treatment of these cases is complicated and the prognosis is extremely poor. We report a case of hepatic echinococcosis in a 40-year-old Tibetan man who was treated with the hepatic right tricuspidectomy + left hepatic duct jejunostomy (Roux-en-Y) surgical procedure. This procedure provides a reference for treatment of similar cases of echinococcosis. For patients with multiorgan metastasis, chemotherapy is the first choice. This should be followed by possible surgical treatment for life-threatening lesions of alveolar echinococcosis and subsequent chemotherapy. Individualized treatment accompanied by multidisciplinary treatment and damage control surgery could optimally benefit patients with advanced hepatic echinococcosis.

目的 探讨肝囊型包虫病手术患者胆漏发生的危险因素,并建立相应的回归预测模型.方法 回顾性分析2014年6月至2018年12月于青海省福利慈善医院行手术治疗的183例肝囊型包虫病患者临床资料.患者均签署知情同意书,符合医学伦理学规定.其中男67例,女116例;年龄12～78岁,中位年龄41岁.采用Logistic回归筛选出胆漏危险因素,森林图显示多因素Logistic回归的各个指标效应大小,建立肝囊型包虫病手术患者胆漏Logistic回归预测模型.通过Hosmer-Lemeshow检验评价预测模型的拟合度,采用受试者工作特征(ROC)曲线评价模型的预测价值.结果 本组患者胆漏发生率为22.4％(41/183).多因素Logistic回归结果显示,ALP>135 U/L(OR=10.797,95％CI:4.860～23.927,P<0.05)、囊肿直径>10 cm(OR=3.576,95％CI:1.911～6.268,P<0.05)是肝囊型包虫病手术患者胆漏发生的独立危险因素.将这两个因素纳入Logistic回归方程:Logit(P)=-2.580+2.412XALP+1.215X囊肿直径.概率模型为:P=1/[1+Exp(-2.580+2.412XALP+1.215X囊肿直径)].ROC曲线下面积为0.792(95％CI:0.719～0.864,P<0.05),敏感度为0.74,特异度为0.70.结论 ALP、囊肿直径是肝囊型包虫病手术患者胆漏发生的危险因素,根据回归分析建立的预测模型能比较有效地预测肝囊型包虫病手术患者胆漏的发生.

目的 探讨七氟醚预处理对藏区肝包虫手术患者肝缺血再灌注损伤(LIRI)和血浆中性粒细胞核转录因子κB(NF-κB)活性的影响.方法 选择就诊的80例肝包虫手术患者,采用随机数字表法将其随机分为藏族世居包虫患者对照组(ZC组)、藏族世居包虫患者七氟醚预处理组(ZS组)、汉族移居包虫患者对照组(HC组)和汉族移居包虫患者七氟醚预处理组(HS组)各20例.ZC组和HC组不用吸入性麻醉药,ZS组和HS组麻醉诱导插管采用七氟醚吸入麻醉.于麻醉诱导前(T0)、肝门阻断即刻(T1)、肝门开放即刻(T2)、缺血/再灌注1 h(T3)、6 h(T4)、24 h(T5),检测各时刻血清丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)、乳酸、肿瘤坏死因子α(TNF-α)、白细胞介素(IL)-6水平和中性粒细胞NF-κB活性;于T3时刻,检测正常肝组织丙二醛(MDA)和超氧化物歧化酶(SOD)水平;HE染色光镜下观察肝组织病理学形态.结果 与ZC组比,ZS组患者在T1～L时间点的ALT、AST、LDH、乳酸、TNF-α、IL-6水平和NF-κB活性均降低(P＜0.05),在T3时间点肝组织的SOD活性升高(P＜0.05),肝组织的MDA水平降低(P＜0.05);与HC组比,HS组患者在T1～T5时间点的ALT、AST、LDH、乳酸、TNF-α、IL-6水平和NF-κB活性均降低(P＜0.05),在T3时间点肝组织的SOD活性升高(P＜0.05),肝组织的MDA水平降低(P＜0.05);HE染色显示ZS组和HS组肝组织病理变化减轻.结论 七氟醚可能通过抑制中性粒细胞NF-κB活性,抑制炎症因子的释放,减轻藏区肝包虫手术患者肝缺血再灌注损伤.

Background: Echinococcosis is a chronic consumptive liver disease. Little research has been carried out on the nutritional status of infected patients, though liver diseases are often associated with malnutrition. Our study investigated four different nutrition screening tools, to assess nutritional risks of hospitalized patients with echinococcosis. Methods: Nutritional Risk Screening 2002 (NRS 2002), Short Form of Mini Nutritional Assessment (MNA-SF), Malnutrition Universal Screening Tool (MUST), and the Nutrition Risk Index (NRI) were used to assess 164 patients with alveolar echinococcosis (AE) and 232 with cystic echinococcosis (CE). Results were then compared with European Society for Clinical Nutrition and Metabolism (ESPEN) criteria for malnutrition diagnosis. Results: According to ESPEN standards for malnutrition diagnosis, 29.2% of CE patients and 31.1% of AE patients were malnourished. The malnutrition risk rates for CE and AE patients were as follows: NRS 2002 – 40.3% and 30.7%; MUST – 51.5% and 50.9%; MNA-SF – 46.8% and 44.1%; and NRI – 51.1% and 67.4%. In patients with CE, MNA-SF and NRS 2002 results correlated well with ESPEN results (k = 0.515, 0.496). Area-under-the-curve (AUC) values of MNA-SF and NRS 2002 were 0.803 and 0.776, respectively. For patients with AE, NRS 2002 and MNA-SF results correlated well with ESPEN (k = 0.555, 0.493). AUC values of NRS 2002 and MNA-SF were 0.776 and 0.792, respectively. Conclusion: This study is the first to analyze hospitalized echinococcosis patients based on these nutritional screening tools. Our results suggest that NRS 2002 and MNA-SF are suitable tools for nutritional screening of inpatients with echinococcosis.

肝囊性病变分为寄生性和非寄生性.寄生性病变主要指肝囊性包虫病,非寄生性病变可再分为良性、交界性和恶性病变.目前对肝囊性占位性病变的规范治疗尚不健全,查阅了国内外相关文献资料,结合文献对肝囊性相关疾病的病因、临床表现、诊断及治疗相关问题进行综述,以提高对本病的认识.

目的 构建适合融合基因pCzn1-TSP3的原核表达系统,纯化获得多房棘球蚴蛋白TSP3.方法 TSP3的基因序列在GenBank中获取,采用基于PAS(PCR-based Accurate Synthesis)的方法,构建原核表达质粒pCzn1-TSP3,并转化于大肠杆菌Arctic Express (DE3)中,经IPTG诱导,获得目的蛋白,利用Ni-IDA-Sepharose CL-6B亲和层析柱纯化获得目的蛋白.结果 经生物公司测序及酶切鉴定结果证明重组质粒pCzn1-TSP3构建成功.进一步实验结果表明TSP3在原核表达系统中主要以可溶蛋白形式存在.继而经Ni-IDA-Sepharose CL-6B亲和层析柱纯化获得TSP3纯蛋白,并经过Western Blot证实.结论 大肠杆菌Arctic Express(DE3)中能够成功表达蛋白TSP3,且Ni-IDA-Sepharose CL-6B亲和层析柱能够纯化目的蛋白TSP3.

Objective Constructed a prokaryotic expression system for CTB_Emy162 and obtained Purified fu-sion proteinCTB_Emy162 in Echinococcus multilocularis.Method According to the sequence of Echinococcus multi-locularis Emy162 antigen in GenBank,Bioinformatics software OptimumTM Codon was used to obtain a suitable se-quence of gene expressed in E.coli BL_21 by optimizing of codon adaptation index(CAI).The optimal codon fre_quency(FOP)and the content of GC.and the optimization sequence were cloned into prokaryotic expression vector and pET_28a.IPTG induction,Ni_NTA affinity chromatography and ion exchange chromatogra_phy were utilized to obtain CTB_Emy162 protein.Results Emy162 sequencewe procured was suitable for prokaryotic expression system by OptimumTM Codon software.Recombinant plasmids pET28a_CTB_Emy162 were constructed successfully and i-dentified by double_enzyme digestion and sequencing.Further,our results were indicated that CTB_Emy162 was ex-pressed in inclusion bodies part of pET28a_CTB_Emy162.Finally,we obtained highly purified recombinant protein CTB_Emy162 by Ni_chelating affinity chromatography.Conclusions In this study,we successfully establish the pro-karyotic expression system of CTB_Emy162,and obtain highly purified CTB_Emy162.

Objective The present study aims to predict the secondary structure and the T_and B_cell epitopes for the Echinococcus multilocularis TSP3 antigen,in order to reveal the dominant epitopes of the antigen.Methods The secondary structure of the protein was analyzed using SOPMA server.The T_cell and B_cell epitopes of TSP3 were predicted using IEDB,Syfpeithi,Bcepred and ABCpred online software.The characteristics of hydrophilicity, flexibility,antigenic propensity and exposed surface area were predicted.Results The random coils and β sheets ac-counted for 25.68% and 4.05% of the secondary structure of the TSP3 protein,respectively.This was indicative of the presence of potential dominant antigenic epitopes in TSP3.Following bioinformatic analysis,numerous distinct antigenic epitopes of TSP3 were identified.The high_scoring T_cell epitopes were located at positions T33_42, T45_55,T53_63,T68_77,T80_90,T92_104,T110_122 and T134_144;whilst the likely B_cell epitopes were lo-cated at positions T18_33,T45_55,T53_63,T64_75,T80_90,T92_104 and T110_122.Conclusions Eight T cell and seven B cell dominant epitopes of the TSP3 antigen were revealed by the bioinformatic methods,which may be useful for the development of a dominant epitope vaccine.

目的 回顾性研究我国肝外包虫病的分布、临床症状特点、诊断及治疗概况.方法 利用中国知网和万方数据库检索2000 ～ 2015年中国(不包括港澳台地区)肝外包虫病的相关病例报道并进行回顾性分析总结.结果 纳入2000 ～ 2015年发表的肝外包虫病相关文献66篇共884例病例,其中分布以新疆最多(68.73％),其次为西藏(8.77％)、宁夏(6.75％)、青海(6.41％)、甘肃(5.62％)几个地区;器官分布以肺最多(61.09％),其次为骨(9.95％)、脑(9.73％)、脾(8.60％)、肾(6.22％)等居多;患者临床症状主要以一般占位症状及包虫所在器官功能异常为主;行影像学及免疫学检查者最多,X线(30.77％)、B超(7.47％)、CT(17.43％)、MR(1.59％)、实验室检查(36.31％);临床治疗大部分患者采取外科手术(73.30％),部分患者辅以药物化疗.结论 我国肝外包虫病的分布与包虫病整体分布大致相同,其肝外分布多在血供丰富的器官,临床诊断以影像学及免疫学检查为主,手术仍为其治疗的主要手段.

The effects of different concentration treatments on botanical and yield characters of Salvia splendens were studied by spraying PP333at the stage of early blooming.The results showed,under concentration of 150 mg/L and 200 mg/L,the height were restrained remarkably,crown were decreased,number of shoot were increased.Under concentration of 100 mg/L and 150 mg/L,the Seed-setting rate and seed yield were remarkably increased,but had no effects on 1000-grain weight(TGW).