Metastatic stomach lymphoepithelioma-like carcinoma and immune checkpoint inhibitor therapy:A case report

BACKGROUND Pulmonary lymphoepithelioma-like carcinoma(PLELC)is a rare type of nonsmall-cell lung cancer.Stomach lymphoepithelioma-like carcinoma(LELC)metastasis secondary to PLELC has not been reported recently.CASE SUMMARY A 64-year-old female was admitted to our hospital for a regular gastroscopy examination with a 6-year history of surgical resection for left PLELC.Positron emission tomography/computed tomography suggested high accumulation of 18F-fludeoxyglucose in the gastric cardia region.Upper gastrointestinal endoscopy confirmed a large mass at the stomach fundus.Immunohistochemistry(IHC)of the biopsy suggested metastatic stomach LELC.Proximal gastrectomy showed that this 6.5 cm×5.0 cm mass was located in the stomach fundus near the cardia.Histopathological examination showed a poorly differentiated carcinoma with prominent lymphoplasmacytic infiltration.IHC demonstrated that the tumor was positive for CK(AE1/AE3),p63,p40,p53,Ki-67(70%),and EGFR(3+)and negative for CK7,CK20,Her2,and CD10.In situ hybridization analysis showed positive staining Epstein-Barr virus-encoded RNA.Tumor programmed cell death ligand 1(PD-L1)expression score was 98%,and the combined positive score was 100,with no evidence of microsatellite instability.Thus,the patient was unequivocally diagnosed with metastatic stomach LELC secondary to pulmonary LELC.After discharge,this patient underwent PD-1 inhibitor treatment(toripalimab,240 mg)every 3 wk for ten cycles,and she has had no tumor recurrence.CONCLUSION For gastric LELC metastasis,PD-1 inhibitor therapy could become a new therapeutic approach,though there is still no evidence from large data sets to support this.

Efficacy of continuous gastric artery infusion chemotherapy in relieving digestive obstruction in advanced gastric cancer

BACKGROUND Obstruction or fullness after feeding is common in gastric cancer(GC)patients,affecting their nutritional status and quality of life.Patients with digestive obstruction are generally in a more advanced stage.Existing methods,including palliative gastrectomy,gastrojejunostomy,endoluminal stent,jejunal nutrition tube and intravenous chemotherapy,have limitations in treating these symptoms.AIM To analyze the efficacy of continuous gastric artery infusion chemotherapy(cGAIC)in relieving digestive obstruction in patients with advanced GC.METHODS This study was a retrospective study.Twenty-nine patients with digestive obstruction of advanced GC who underwent at least one cycle of treatment were reviewed at The Second Affiliated Hospital of Zhejiang University School of Medicine.The oxaliplatin-based intra-arterial infusion regimen was applied in all patients.Mild systemic chemotherapy was used in combination with local treatment.The clinical response was evaluated by contrast-enhanced computed tomography using Response Evaluation Criteria In Solid Tumors(RECIST)criteria.Digestive tract symptoms and toxic effects were analyzed regularly.A comparison of the Karnofsky Performance Status(KPS)score and Stooler’s Dysphagia Score before and after therapy was made.Univariate survival analysis and multivariate survival analysis were also performed to explore the key factors affecting patient survival.RESULTS All patients finished cGAIC successfully without microcatheter displacement,as confirmed by arteriography.The median follow-up time was 24 mo(95%CI:20.24-27.76 mo).The overall response rate was 89.7%after cGAIC according to the RECIST criteria.The postoperative Stooler’s Dysphagia Score was significantly improved.Twentytwo(75.9%)of the 29 patients experienced relief of digestive obstruction after the first two cycles,and 13(44.8%)initially unresectable patients were then considered radically resectable.The median overall survival time(mOS)was 16 mo(95%CI:9.32-22.68 mo).Patients who received radical surgery had a significantly longer mOS than other patients(P value<0.001).Multivariate Cox regression analysis indicated that radical resection after cGAIC,intravenous chemotherapy after cGAIC,and immunotherapy after cGAIC were independent predictors of mOS.None of the patients stopped treatment because of adverse events.CONCLUSION cGAIC was effective and safe in relieving digestive obstruction in advanced GC,and it could improve surgical conversion possibility and survival time.

内科胸腔镜治疗结核性胸膜炎的意义

目的分析内科胸腔镜治疗结核性胸膜炎的临床意义。方法收集该院40例诊断为结核性胸膜炎的患者,其中由内科胸腔镜下确诊结核性胸膜炎患者为20例,纳为治疗组;由常规胸腔穿刺后通过胸水化验、结核菌素试验而临床诊断为结核性胸膜炎患者20例,纳为对照组;两组诊断后均给予了相同抗结核方案治疗。计算两组患者住院天数,统计治疗组、对照组住院期间共行的胸腔穿刺数;测量两组患者治疗12个月后的胸部CT上胸膜肥厚度。结果治疗组和对照组的平均住院天数分别为(9.55±1.63)和(13.60±3.59)d,治疗组明显短于对照组,两组比较差异有统计学意义(P<0.05);治疗组与对照组平均行胸腔穿刺术次数为(1.50±0.76)和(2.80±0.99)次,治疗组明显少于对照组,两组比较差异有统计学意义(P<0.05)。治疗组与对照组治疗12个月后的胸膜CT上平均胸膜肥厚度分别为(0.15±0.09)和(2.30±0.13)mm,治疗组明显薄于对照组,两组比较差异有统计学意义(P<0.05)。结论使用内科胸腔镜下确诊结核性胸膜炎的患者,可减少住院日,减少胸腔穿刺频次,减少胸膜肥厚度。临床上若疑诊结核性胸膜炎者,建议优先考虑行内科胸腔镜诊治。

Study of transactivating effect of pre-S2 protein of hepatitis B virus and cloning of genes transactivated by pre-S2 protein with suppression subtractive hybridization

AIM: To investigate the transactivating effect of pre-S2 protein of hepatitis B virus (HBV) and construct a subtractive cDNA library of genes transactivated by pre-S2 protein with suppression subtractive hybridization (SSH) technique, and to pave the way for elucidating the pathogenesis of HBV infection. METHODS: pcDNA3.1(-)-pre-S2 containing pre-S2 region of HBV genome was constructed by routine molecular methods. HepG2 cells were cotransfected with pcDNA3.1 (-)-pre-S2/pSV-lacZ and empty pcDNA3.1(-)/pSV-lacZ. After 48 h, cells were collected and detected for the expression of β-galactosidase (β-gal). SSH and bioinformatics techniques were used, the mRNA of HepG2 cells transfected with pcDNA3.1(-)-pre-S2 and pcDNA3.1(-) empty vector was isolated, respectively, cDNA was synthesized. After digestion with restriction enzyme RsaI, cDNA fragments were obtained. Tester cDNA was then divided into two groups and ligated to the specific adaptor 1 and adaptor 2, respectively. After tester cDNA was hybridized with driver cDNA twice and underwent two times of nested PCR, amplified cDNA fragments were subcloned into pGEM-Teasy vectors to set up the subtractive library. Amplification of the library was carried out with E.coli strain DH5oα. The cDNA was sequenced and analyzed in GenBank with Blast search after PCR. RESULTS: The pre-S2 mRNA could be detected in HepG2 cells transfected with pcDNA3.1(-)-pre-S2 plasmid. The activity of β-gal in HepG2 cells transfected with pcDNA3.1 (-)-pre-S2/pSV-lacZ was 7.0 times higher than that of control plasmid (P<0.01). The subtractive library of genes transactivated by HBV pre-S2 protein was constructed successfully. The amplified library contains 96 positive clones. Colony PCR showed that 86 clones contained 200-1 000 bp inserts. Sequence analysis was performed in 50 clones randomly, and the full length sequences were obtained with bioinformatics method and searched for homologous DNA sequence from GenBank, altogether 25 coding sequences were obtained, these cDNA sequences might be the target genes transactivated by pre-S2 protein. CONCLUSION: The pre-S2 protein of HBV has transactivating effect on SV40 early promoter. The obtained sequences may be target genes transactivated by pre-S2 protein among which some genes coding proteins involved in cell cycle regulation, metabolism, immunity, signal transduction and cell apoptosis.This finding brings some new clues for studying the biological functions of pre-S2 protein and further understanding of HBV hepatocarcinogesis.

Hepatocellular Carcinoma with Hypersplenic Thrombocytopenia and Situs Inversus Totalis:A Case Report

SITUS inversus totalis （SIT） is a complete mirrorimage of the thoracic and abdominal viscera,occurring at an incidence of 1 in 5000-20 000live births.1, 2 It is supposed to originate from anabnormal rotation of the cardiac tube during embryogenesis.Although SIT is a congenital anomaly, most of patients aredetected accidentally at the time of radiological investigation.Hepatocellular carcinoma （HCC） is usually associated withliver cirrhosis and portal hypertension in Chinese. Presenceof hypersplenic thrombocytopenia is a common consequenceof long-term portal hypertension in cirrhoticpatients.

A young male with abdominal distention and fever after cardiopulmonary resuscitation treatment

Abdominal distention after cardiopulmonary resuscitation(CPR)is a common phenomenon that presents in the emergency department.It is caused by a long period of bag-valve mask ventilation or esophageal intubation.However,a rare but life-threating diagnosis should be considered when distention progresses rapidly.Here,we reported a patient who developed a fever and abdominal pain that couldn’t be relieved by gastric decompression.

Association of the Cellular Apoptosis Susceptibility Protein with HBV Infection in Hepatocellular Carcinoma

Objective The cellular apoptosis susceptibility(CAS) protein plays a regulatory role in the induction of cell death in tumor cells. The objective of this study was to investigate the association of the expression of CAS protein with HBV infection in the development of HCC. Methods The expression level of CAS was measured with immunohistochemistry. The occurrence of HBs Ag, HBe Ag and HBV DNA in HCC were concurrently examined with immunohistochemistry and in situ hybridization, respectively. Results The results showed that the CAS protein was detected in 86%(43/50), 70%(7/10), 15%(3/20) and none(0/20) of livers from patients with HCC, cholangiocarcinoma, cirrhosis and hepatitis, respectively. Furthermore, the level of CAS protein was higher in poorly differentiated tumors than moderately or well differentiated HCC. Interestingly, the CAS was stained significantly stronger in HBV-infected HCC than in non-HBV infected tissues(P < 0.01). Conclusions The expression of CAS is facilitated by HBV infection in HCC, suggesting that CAS might be a prognostic marker and a putative therapeutic target for HCC.

Epigenetic Silencing of Eyes Absent 4 Gene by Acute Myeloid Leukemia 1-Eight-twenty-one Oncoprotein Contributes to Leukemogenesis in t（8;21） Acute Myeloid Leukemia

Background： The acute myeloid leukemia 1 （AML1）-eight-twenty-one （ETO） fusion protein generated by the t（8;2 1）（q22;q22） translocation is considered to display a crucial role in leukemogenesis in AML. By focusing on the anti-leukemia effects of eyes absent 4 （EYA4） gene on AML cells, we investigated the biologic and molecular mechanism associated with AML 1 -ETO expressed in t（8;21） AML. Methods： Qualitative polymerase chain reaction （PCR）, quantitative reverse transcription PCR （RT-PCR）, and Western blotting analysis were used to observe the mRNA and protein expression levels of EYA4 in cell lines. Different plasmids （including mutant plasmids） of dual luciferase reporter vector were built to study the binding status of AML1-ETO to the promoter region of EYA4. Chromatin immunoprecipitation assay was used to study the epigenetic silencing mechanism of EYA4. Bisulfite sequencing was applied to detect the methylation status in EYA4 promoter region. The influence ofEYA4 gene in the cell proliferation, apoptosis, and cell clone-forming ability was detected by the technique of Cell Counting Kit-8, flow cytometry, and clonogenic assay. Results： EYA4 gene was hyperrnethylated in AMLI-ETO＋ patients and its expression was down-regulated by 6-fold in Kasumi-1 and SKNO-1 cells, compared to HL-60 and SKNO-1-siA/E cells, respectively. We demonstrated that AML1-ETO triggered the epigenetic silencing of EYA 4 gene by binding at AML 1-binding sites and recruiting histone deacetylase 1 and DNA methyltransferases. Enhanced EYA4 expression levels inhibited cellular proliferation and suppressed cell colony formation in AMLI-ETO cell lines. We also found EYA4 transfection increased apoptosis of Kasumi- 1 and SKNO-1 cells by 1.6-fold and 1.4-fold compared to negative control, respectively. Conclusions： Our study identified EYA4 gene as targets for AML1-ETO and indicated it as a novel tumor suppressor gene. In addition, we provided evidence that EYA4 gene might be a novel therapeutic target and a potential candidate for treating AML 1-ETO＋ t （8;21 ） AML.