Highly expressed carbohydrate sulfotransferase 11 correlates with unfavorable prognosis and immune evasion of hepatocellular carcinoma.

Despite great advance has been made in multi-modality treatments for HCC patients, the effectiveness is far from satisfactory with worse survival outcome, which may be partly explainable by the anti-tumor deficiency of the immune system. It is necessary to clarify the molecular mechanism of HCC immunodeficiency. Here, we demonstrated that carbohydrate sulfotransferase 11 (CHST11) was upregulated in HCC and related to advanced TNM stage. HCC patients with TP53 mutation showed higher CHST11 expression. Survival analysis revealed that CHST11 was an independent prognostic biomarker in HCC. Cellular functional experiments indicated that knockdown of CHST11 in HCC inhibited cell proliferation and metastasis. Gene functional enrichment analyses indicated that CHST11 modulated pathways related to tumor growth, metastasis and immune regulation. Continuative immune-related analyses revealed that CHST11 expression facilitated Tregs infiltration in HCC and promoted the expression of checkpoints PD-L1/PD-1, resulting in the immunosuppression of HCC. Targeting CHST11 may inhibit Tregs infiltration and enhance the antineoplastic effect of immune checkpoint inhibitors, which provides a novel insight into the combination immunotherapy with Treg-modulating agents and PD-L1/PD-1 inhibitors.

Five circRNAs Serve as Potential Diagnostic and Prognostic Biomarkers in Lung Adenocarcinoma.

BACKGROUND: The mortality rate of lung adenocarcinoma ranks first worldwide, higher than gastric, colorectal, breast, and other cancers. The lack of effective and accurate diagnosis contributes to the patient's unfavorable prognosis with lung adenocarcinoma since most patients are diagnosed at a late stage. In the present study, we aimed to investigate five circRNAs correlated with lung adenocarcinoma and their clinical roles. METHODS: We collected 68 unpaired serum and tissue samples from patients with lung adenocarcinoma and healthy volunteers. At the next stage, quantitative real-time polymerase chain reaction (qRT-PCR) assays were conducted. Furthermore, we uncovered the correlation of their expressions with clinicopathological features and the diagnostic values. Finally, the 5-year survival rate and disease-free rate were analyzed using Kaplan-Meier methods. RESULTS: The results revealed that expression levels of hsa_circ_001010, hsa_circ-ZNF609 were significantly elevated while hsa_circ-CRIMI1, hsa_circ-EPB41L2, and hsa_circ_0072309 were lower in lung adenocarcinoma serum samples and tissues than those in healthy controls (p < 0.05). Meanwhile, hsa_circ_001010 and hsa_circ-ZNF609 were downregulated hsa_circ-CRIMI1, hsa_circ-EPB41L2, and hsa_circ_0072309 were elevated in A549 cells compared with BEAS-2B cells. Knockdown of hsa_circ-CRIMI1, hsa_circ-EPB41L2, or hsa_circ_0072309 and overexpressing of hsa_circ_001010 and hsa_circ-ZNF609 could promote A549 cell apoptosis but inhibits proliferation as well. Furthermore, receiver operating characteristic (ROC) assays demonstrated that the area under the curve (AUC) were as follows: hsa_circ_001010 (0.8512, 95% CI, 0.7872 - 0.9152; p < 0.0001), hsa_circ-ZNF609 (0.7876, 95% CI, 0.7101 - 0.8651; p < 0.0001), hsa_circ-CRIMI1 (0.6614, 95% CI, 0.5708 - 0.7521; p < 0.0001), hsa_circ-EPB41L2 (0.6851, 95% CI, 0.5960 - 0.7742; p = 0.0002), and hsa_circ_0072309 (0.7359, 95% CI, 0.6250 - 0.8199; p < 0.0001). Notably, higher expressions of hsa_circ_001010, hsa_circ-ZNF609, and lower expressions of hsa_circ-CRIMI1, hsa_circ-BGT2, hsa_circ-EPB41L2, and hsa_circ_0072309 were positively correlated with clin-ical stage, lymph node metastasis, and smoking. Last but not least, patients with higher expressions of hsa_ circ_001010, hsa_circ-ZNF609, and lower expressions of hsa_circ-CRIMI1, hsa_circ-EPB41L2, and hsa_circ_ 0072309 had significantly lower overall survival rates and disease-free rates as well. CONCLUSIONS: We concluded that the five circRNAs might have diagnostic and prognosis significance in lung ade-nocarcinoma. However, further functional studies are warranted to ascertain the biological mechanisms of these circRNAs in the occurrence and development of lung adenocarcinoma.

Learning Curve for Uniportal Thoracoscopic Anatomical Pulmonary Segmentectomy.

Background. Video-assisted thoracoscopic segmentectomy has been considered to be an effective management in properly selected patients. Given the substantial technical difficulties, uniportal thoracoscopic segmentectomy is a challenging surgical procedure. The purpose of this study was to investigate the learning curve for uniportal thoracoscopic segmentectomy procedures. Methods. Prospectively collected data were retrospectively analyzed for a series of 124 consecutive patients undergoing uniportal thoracoscopic segmentectomy from August 2016 to September 2018. The learning curve for uniportal thoracoscopic segmentectomy was studied using cumulative sum (CUSUM) combined with risk-adjusted cumulative sum (RA-CUSUM) methods. Results. According to the CUSUM combined with the RA-CUSUM analyses, the learning curve for uniportal thoracoscopic segmentectomy was composed of three well-differentiated phases: cases 1 to 24, cases 25 to 57, and cases 58 to 124. With the procedures proceeded orderly, the operation time tends to progressively decrease. Conclusions. The uniportal thoracoscopic segmentectomy is considered to be a feasible and safe surgical procedure that can be taught to surgeons through a proctored learning curve. The data of the present study indicate that the inflection point for completion the learning curve was reached after a considerable number of 57 cases.

FLCN-regulated miRNAs suppressed reparative response in cells and pulmonary lesions of Birt-Hogg-Dubé syndrome.

BACKGROUND: Birt-Hogg-Dubé Syndrome (BHDS) characterised by skin fibrofolliculomas, kidney tumour and pulmonary cysts/pneumothorax is caused by folliculin (FLCN) germline mutations. The pathology of both neoplasia and focused tissue loss of BHDS strongly features tissue-specific behaviour of the gene. Isolated cysts/pneumothorax is the most frequent atypical presentation of BHDS and often misdiagnosed as primary spontaneous pneumothorax (PSP). Deferential diagnosis of BHDS with isolated pulmonary presentation (PSP-BHD) from PSP is essential in lifelong surveillance for developing renal cell carcinoma. METHODS: The expression profiles of microRNAs (miRNAs) in cystic lesions of PSP-BHD and PSP were determined via microarray. The selected upregulated miRNAs were further confirmed in the plasma of an expanded cohort of PSP-BHD patients by reverse transcription quantitative PCR (RT-qPCR). Their diagnostic accuracy was evaluated. Moreover, the cellular functions and targeted signalling pathways of FLCN-regulated miRNAs were assessed in various cell lines and in the lesion tissue contexts. RESULTS: Cystic lesions of PSP-BHD and PSP showed different miRNAs profiles with a significant upregulation of miR-424-5p and let-7d-5p in PSP-BHD. The combination of the two effectively predicted BHDS patients. In vitro studies revealed a suppressive effect of FLCN on miR-424-5p and let-7d-5p expressions specifically in lung epithelial cells. The ectopic miRNAs triggered epithelial apoptosis and epithelial transition of mesenchymal cells and suppressed the reparative responses in cells and tissues with FLCN deficiency. CONCLUSION: The upregulation of miR-424-5p and let-7d-5p by FLCN deficiency occurred in epithelial cells and marked the PSP-BHD condition, which contributed to a focused degenerative pathology in the lung of PSP-BHD patients.

Modified procedure of uniportal video-assisted thoracoscopic lobectomy with muscle sparing incision.

BACKGROUND: To describe modified procedure for uniportal video-assisted thoracoscopic surgery (UVATS) lobectomy with a small, total muscle-sparing incision. METHODS: Forty-eight UVATS leucotomy were attempted and successfully completed. A single incision of approximately 3 cm was made in an intercostal space along the anterior axillary line. Muscle-sparing technique was applied with this single-incision approach using muscle sparing technique. RESULTS: Incision size was kept to a minimum, with a median of 3 cm. Mediastinal lymph node dissection was performed in all patients with malignancy. Overall median operative time was 1.3 hours. Median hospitalization was 13.5 days (range, 6-21 days). Morbidity rate was low at 3%. There were no other postoperative complications, mortality, or re-admissions. CONCLUSIONS: Modified procedure of lobectomy with UVATS might be easy to operate with less surgical time and morbidity rate, muscle sparing technique might reduce post-operation pain.

A rapid NGS strategy for comprehensive molecular diagnosis of Birt-Hogg-Dubé syndrome in patients with primary spontaneous pneumothorax.

BACKGROUND: Primary spontaneous pneumothorax (PSP) or pulmonary cysts is one of the manifestations of Birt-Hogg-Dube syndrome (BHDS) that is caused by heterozygous mutations in FLCN gene. Most of the mutations are SNVs and small indels, and there are also approximately 10 % large intragenic deletions and duplications of the mutations. These molecular findings are generally obtained by disparate methods including Sanger sequencing and Multiple Ligation-dependent Probe Amplification in the clinical laboratory. In addition, as a genetically heterogeneous disorder, PSP may be caused by mutations in multiple genes include FBN1, COL3A1, CBS, SERPINA1 and TSC1/TSC2 genes. For differential diagnosis, these genes should also be screened which makes the diagnostic procedure more time-consuming and labor-intensive. METHODS: Forty PSP patients were divided into 2 groups. Nineteen patients with different pathogenic mutations of FLCN previously identified by conventional Sanger sequencing and MLPA were included in test group, 21 random PSP patients without any genetic screening were included in blinded sample group. 7 PSP genes including FLCN, FBN1, COL3A1, CBS, SERPINA1 and TSC1/TSC2 were designed and enriched by Haloplex system, sequenced on a Miseq platform and analyzed in the 40 patients to evaluate the performance of the targeted-NGS method. RESULTS: We demonstrated that the full spectrum of genes associated with pneumothorax including FLCN gene mutations can be identified simultaneously in multiplexed sequence data. Noteworthy, by our in-house copy number analysis of the sequence data, we could not only detect intragenic deletions, but also determine approximate deletion junctions simultaneously. CONCLUSIONS: NGS based Haloplex target enrichment technology is proved to be a rapid and cost-effective screening strategy for the comprehensive molecular diagnosis of BHDS in PSP patients, as it can replace Sanger sequencing and MLPA by simultaneously detecting exonic and intronic SNVs, small indels, large intragenic deletions and determining deletion junctions in PSP-related genes.

Bronchoplasty using continuous suture in complete monitor view: a suitable method of thoracoscopic sleeve lobectomy for non-small cell lung cancer.

BACKGROUND: Our study aims to determine the value of bronchial anastomosis using complete continuous suture. METHODS: Six patients diagnosed with central lung carcinoma who were candidates for right-sided sleeve lobectomy and underwent sleeve resection of the right upper lobe by thoracoscopic surgical procedure. RESULTS: The mean surgical time was 182 min (range, 110 to 260 min). The mean time of bronchial anastomosis was 49 min (range, 18 to 76 min). The mean bleeding was 110 mL (range, 50 to 260 mL). Median chest tube drainage was 305 mL (range, 200 to 600 mL). No perioperative deaths or major complications occurred. The postoperative bronchoscopy confirmed no stenosis. The mean follow-up time was 19.2 months (range, 7 to 34 months), and six patients were alive. CONCLUSIONS: Bronchial anastomosis using complete continuous suture may be a suitable method in thoracoscopic sleeve lobectomy.

Localization of Small Peripheral Pulmonary Lesion by Methylene Blue Injection With Radial Endobronchial Ultrasonography in Sublobar Resection.

Ultrasonography can be performed as an adjunct to aid in the localization of small nodules. We describe 25 patients with computed tomographic evidence of peripheral pulmonary lesions (PPLs) who underwent bronchoscopy with endobronchial ultrasonography (EBUS) for localization. The results demonstrate that methylene blue injection with radial EBUS is a simple and easy location technique for PLLs in Sublobar resection.

Right lung cancer with partial anomalous pulmonary venous return and absent right upper lobe.

When a vascular shunt is present, major lung resection may cause acute right heart failure. We report a case of right lung cancer with ipsilateral partial anomalous pulmonary venous return (PAPVR) and absent right upper lobe managed by pneumonectomy. A 48-year-old former smoker was diagnosed as right lung cancer; chest computed tomography (CT) revealed an anomalous right pulmonary vein draining into inferior vena cava. Bronchus of right upper lobe was not found in fiberoptic bronchoscopy. Right pneumonectomy was performed. Pathological examination revealed a T2aN2M0 squamous lung cancer. The patient tolerated well without notable descending of arterial oxygen pressure.

Promoter methylation is not associated with FLCN irregulation in lung cyst lesions of primary spontaneous pneumothorax.

Germline mutations in FLCN are responsible for ~10% of patients with primary spontaneous pneumothorax (PSP), characterized by multiple lung cysts in the middle/lower lobes and recurrent pneumothorax. These clinical features are also observed in a substantial portion of patients with sporadic PSP exhibiting no FLCN coding mutations. To assess the potential underlying mechanisms, 71 patients with PSP were selected, including 69 sporadic and 2 familial cases, who bared FLCN mutation­like lung cysts, however, harbored no FLCN protein­altering mutations. Notably, in a significant proportion of the patients, FLCN irregulation was observed at the transcript and protein levels. Genetic analyses of the cis­regulatory region of FLCN were performed by sequencing and multiplex ligation­dependent probe amplification assay. No inheritable DNA defect was detected, with the exception of a heterozygous deletion spanning the FLCN promoter, which was identified in a family with PSP. This mutation caused a reduction in the expression of FLCN in the lung cysts. Pedigree analysis demonstrated that haploinsufficiency of FLCN was pathogenic. To determine whether epigenetic mechanisms may be involved in the irregulation of FLCN, the promoter methylation status was measured in the remainder of the patients. No evidence of FLCN promoter methylation was demonstrated. The present study suggested that FLCN irregulation in lung cysts of PSP is not associated with promoter methylation.

Controlled Trachea Suspension for Tracheomalacia After Resection of Large Anterior Mediastinal Mass.

Tracheomalacia is a disorder of the large airways that is often caused by a large anterior mediastinal mass. This study describes 7 patients who underwent controlled trachea suspension as a surgical intervention to prevent severe tracheomalacia and provide potent relief of airway symptoms. All patients recovered well. The results demonstrate this procedure may be safe and effective for resection of a large mediastinal mass compressing the trachea with collapsed segments.

Ipsilateral partial anomalous pulmonary venous connection in right lung cancer with absent right upper lobe.

A partial anomalous pulmonary venous connection (PAPVC) is an uncommon congenital anomaly. This report documents the case of a 48-year-old man with PAPVC which was incidentally discovered with right lung cancer and absence of right upper lobe. Right pneumonectomy was successfully performed, and the patient had an uneventful postoperative course. Asymptomatic PAPVC without septal defect is extremely rare. If the PAPVC is located in a different lobe, a pulmonary resection for lung cancer would precipitate an adverse outcome without a correction of the PAPVC. Surgeons should therefore be cautious regarding the potential existence of a PAPVC when a patient undergoes surgical procedures.

FLCN intragenic deletions in Chinese familial primary spontaneous pneumothorax.

Primary spontaneous pneumothorax (PSP) is a significant clinical problem, affecting tens of thousands patients annually. Germline mutations in the FLCN gene have been implicated in etiology of familial PSP (FPSP). Most of the currently identified FLCN mutations are small indels or point mutations that detected by Sanger sequencing. The aim of this study was to determine large FLCN deletions in PSP families that having no FLCN sequence-mutations. Multiplex ligation-dependent probe amplification (MLPA) assays and breakpoint analyses were used to detect and characterize the deletions. Three heterozygous FLCN intragenic deletions were identified in nine unrelated Chinese families including the exons 1-3 deletion in two families, the exons 9-14 deletion in five families and the exon 14 deletion in two families. All deletion breakpoints are located in Alu repeats. A 5.5 Mb disease haplotype shared in the five families with exons 9-14 deletion may date the appearance of this deletion back to approximately 16 generations ago. Evidences for founder effects of the other two deletions were also observed. This report documents the first identification of founder mutations in FLCN, as well as expands mutation spectrum of the gene. Our findings strengthen the view that MLPA analysis for intragenic deletions/duplications, as an important genetic testing complementary to DNA sequencing, should be used for clinical molecular diagnosis in FPSP.

Emergency operation of a patient with spontaneous rupture and massive hemorrhage of pleural solitary fibrous tumor.

Solitary fibrous tumor (SFT) is a rare pleural disease with asymptomatic clinical course. We report a case of a patient with spontaneous rupture and massive hemorrhage of pleural SFT, which was treated by emergency operation.

Primitive chest wall neuroectodermal tumor in a pediatric patient.

A 13-year-old boy with a primitive neuroectodermal tumor of the chest wall is presented. After four cycles of chemotherapy, a computed tomography scan of his chest showed a larger mass invading the left upper lobe of the lung. He underwent resection of the left chest wall from the left fourth to sixth ribs, including the tumor, combined with left upper lobectomy and lymph node dissection. A diagnosis of primitive neuroectodermal tumor was confirmed histopathologically and immunohistochemically. After surgery, four cycles of chemotherapy with ifosfamide and etoposide were given. One year after treatment, the patient is currently doing well without evidence of recurrence.

[Bronchial sleeve lobectomy and carinal resection in the treatment of central lung cancer: a report of 92 cases].

BACKGROUND: Surgery is the best treatment for early and middle stage non-small cell lung cancer. The aim of this study is to summarize the experience of bronchial sleeve mortality lobectomy and carinal resection in the treatment of 92 patients with central lung cancer from January, 1996 to May, 2010. METHODS: A total of 92 patients with central lung cancer underwent pulmonary resection. Carinal resection and reconstruction were performed in 14 patients, bronchial sleeve resection in 70 patients, and bronchial sleeve combined with pulmonary artery sleeve lobectomy in 8 patients. RESULTS: There was no operative mortality. The average operation time was 2 hours and 43 minutes. Postoperative complications such as pulmonary atelectasis occurred in 6.94% (7/92) of total group, and hoarseness in 4.35% (4/92). The 1, 3 and 5 year survival rates were 80.7%, 59.6% and 31.5%. CONCLUSIONS: Bronchial sleeve lobectomy and double sleeve lobectomy are capable of excising pulmonary tumor as much as possible while remaining healthy lung tissues. Carinal resection and reconstruction is helpful to extend the surgical indication, and increase the chance of successful resection.