Ganonorsterone A, a norsteroid from the medicinal fungus Ganoderma lingzhi.

Phytochemical investigation on the fruiting bodies of the medicinal fungus Ganoderma lingzhi led to the isolation of a new norsteroid, namely ganonorsterone A (1), together with one known steroid, cyathisterol (2). The structure and absolute configuration of compound 1 were assigned by extensive analysis of MS, NMR data, and quantum-chemical calculations including electronic circular dichroism (ECD) and calculated 13C NMR-DP4+ analysis. Bioassay results showed that compound 1 displayed moderate inhibition on NO production in RAW 264.7 macrophages.

Mutation spectrum of FLT3 and significance of non-canonical FLT3 mutations in haematological malignancy.

Fms-like tyrosine kinase 3 (FLT3) is frequently mutated in haematological malignancies. Although canonical FLT3 mutations including internal tandem duplications (ITDs) and tyrosine kinase domains (TKDs) have been extensively studied, little is known about the clinical significance of non-canonical FLT3 mutations. Here, we first profiled the spectrum of FLT3 mutations in 869 consecutively newly diagnosed acute myeloid leukaemia (AML), myelodysplastic syndrome and acute lymphoblastic leukaemia patients. Our results showed four types of non-canonical FLT3 mutations depending on the affected protein structure: namely non-canonical point mutations (NCPMs) (19.2%), deletion (0.7%), frameshift (0.8%) and ITD outside the juxtamembrane domain (JMD) and TKD1 regions (0.5%). Furthermore, we found that the survival of patients with high-frequency (>1%) FLT3-NCPM in AML was comparable to those with canonical TKD. In vitro studies using seven representative FLT3-deletion or frameshift mutant constructs showed that the deletion mutants of TKD1 and the FLT3-ITD mutant of TKD2 had significantly higher kinase activity than wild-type FLT3, whereas the deletion mutants of JMD had phosphorylation levels comparable with wild-type FLT3. All tested deletion mutations and ITD were sensitive to AC220 and sorafenib. Collectively, these data enrich our understanding of FLT3 non-canonical mutations in haematological malignancies. Our results may also facilitate prognostic stratification and targeted therapy of AML with FLT3 non-canonical mutations.

Combination of Venetoclax and Midostaurin Efficiently Suppressed Relapsed t(8;21)Acute Myeloid Leukemia With Mutant KIT After Failure of Venetoclax Plus Azacitidine Treatment.

Acute myeloid leukemia (AML) with t(8;21) is categorized as favorable-risk AML, but KIT mutations show a significantly poor prognostic impact in such patients. Persistent vulnerability to relapse is a major challenge in the treatment of this subtype of patients. Venetoclax is a BCL-2 selective inhibitor. The venetoclax+HMA strategy is also a notable salvage regimen that achieves good clinical outcomes in the treatment of relapsed or refractory (R/R) AML. However, in our clinical practice, we found that disease progressed rapidly even after venetoclax+azacitidine (AZA) therapy in two relapsed t(8;21) AML patients with KIT mutations. We report for the first time the therapeutic potential of venetoclax+midostaurin as a new combination therapy for relapsed t(8;21) AMLs with KIT mutations showing resistance to venetoclax+AZA therapy. Our ex vivo study also showed that midostaurin alone could inhibit proliferation and induce apoptosis of Kasumi-1 cells (e.g. Midostaurin induced G2 phase cell arrest, down-regulated p-KIT and BCL-2, while Bax protein levels were up-regulated) and observed a synergistic anti effect when the two drugs were combined. Our study shows that the venetoclax+midostaurin regimen may be a promising treatment option for R/R t(8;21) AML with KIT mutations.

The Crucial Role of Wntless in Osteogenesis and Odontogenesis.

Wnt signalling pathways have been the focus of intense research activity for decades due to their fundamental role in skeletal and dental development. Wntless, an exclusive chaperone protein for the exocytotis of Wnt ligands, was identified in 2006. In the last decade, the molecular biological studies of Wntless and its genetic studies in human and mice have highlighted the importance of this protein in mineralised tissues, including bone, cartilage and teeth. This article reviews recent developments and discrepancies in the role of Wntless in skeletal and dental development based on mutant phenotypes, as well as the underlying mechanism involved in its molecular and physiological regulation. We conclude that, though some controversial phenotypes exist due to different Cre line resources, Cre recombinase activity and detection time points, Wntless undeniably exerts a context- and stage-dependent regulatory function during the development and homeostasis of both skeletal and dental tissue.

FEN1 inhibitor increases sensitivity of radiotherapy in cervical cancer cells.

BACKGROUND: Cervical cancer is one of the most common causes of cancer-associated mortality among affected women in the world. At present, treatment with weekly cisplatin plus ionizing radiation (IR) therapy is the standard regimen for cervical cancer, especially for locally advanced cervical cancer. The purpose of this study is to determine whether FEN1 inhibitors could enhance the therapeutic effect of IR therapy. METHODS: Western blot was applied to determine the expression of FEN1- and apoptosis-related proteins. Cell growth inhibition assay and colony formation assay were used to determine the effects of FEN1 inhibitor and IR exposure for Hela cells in vitro. CRISPR technology was used to knockdown FEN1 expression level of 293T cells, and tumor xenograft in nude mice was employed to determine the effects of FEN1 inhibitor and IR exposure on tumor growth in vivo. RESULTS: Our data revealed that FEN1 is overexpressed in HeLa cell and can be upregulated further by IR. We also demonstrated that FEN1 inhibitor enhances IR sensitivity of cervical cancer in vitro and in vivo. CONCLUSION: FEN1 inhibitor SC13 could sensitize radiotherapy of cervical cancer cell.

Transcriptome Analysis of FEN1 Knockdown HEK293T Cell Strain Reveals Alteration in Nucleic Acid Metabolism, Virus Infection, Cell Morphogenesis and Cancer Development.

AIM AND OBJECTIVE: Flap endonuclease-1 (FEN1) plays a central role in DNA replication and DNA damage repair process. In mammals, FEN1 functional sites variation is related to cancer and chronic inflammation, and supports the role of FEN1 as a tumor suppressor. However, FEN1 is overexpressed in multiple types of cancer cells and is associated with drug resistance, supporting its role as an oncogene. Hence, it is vital to explore the multi-functions of FEN1 in normal cell metabolic process. This study was undertaken to examine how the gene expression profile changes when FEN1 is downregulated in 293T cells. MATERIALS AND METHODS: Using the RNA sequencing and real-time PCR approaches, the transcript expression profile of FEN1 knockdown HEK293T cells have been detected for the next step evaluation, analyzation, and validation. RESULTS: Our results confirmed that FEN1 is important for cell viability. We showed that when FEN1 downregulation led to the interruption of nucleic acids related metabolisms, cell cycle related metabolisms are significantly interrupted. FEN1 may also participate in non-coding RNA processing, ribosome RNA processing, transfer RNA processing, ribosome biogenesis, virus infection and cell morphogenesis. CONCLUSION: These findings provide insight into how FEN1 nuclease might regulate a wide variety of biological processes, and laid the foundation for understanding the role of other RAD2 family nucleases in cell growth and metabolism.

[Mutation Spectrum of FANCJ Gene in Adult Acute Myeloid Leukemia].

OBJECTIVE: To detect and analyze the mutation status of FANCJ gene in adult AML patients, so as to provide the basis for studying the mechanism of FANCJ driven AML and guiding the preventim and treatment of deseese. METHODS: The cDNAs were extracted and transeripted from bone marrow cells and normal skin cells in 222 newly diagnosed AML patients. The primers were designed for FANCJ gene coding region, the mutations of FANCJ gene coding region in AML patients as well as the mutations of FANCJ gene in mucous membrane epethelia in patients were detected by PCR and sanger seguencing; the evolutionary conservation of FANCJ mutation in different organisms was analyzed by NCBI Blast online bioinformaties software. RESULTS: The sequencing analysis showed that the mutations of FANCJ gene happened in 11 sites of FANCJ gene coding region, which were as followed: exon5:c.G430A:p.A144T, exon6:c.A587G:pN196S, exon9:c.C1255T:p.R419W, exon10:c.G1442A:p.G481D, exon11:c.C1609G:p.L537V, exon16:c.C2360T:p.P787L, exon17:c.C2440T:p.R814C, exon19:c.C2608T:pH870Y, exon19:c.A2686G:p.I896V, exon19:c.C2830G:p.Q944E, exon20:c.G3412A:p.D1138N. Among them, the repeatability existed in mutations of A144T, N196S, R814C, I896V and Q944E. Beside, the mutation sites of A144, R419, G381, L537, P787, H870, Q944 and D1138 were highly conserved in different organisms. CONCLUSION: Among 222 adult AML patients, the mutations of FANCJ gene have been found in 26 patients, moreover, the mutation sites are relatively conserved in different organisms, and possess important fanction. The results of this study provide the basis for exploring the mexhanism of FANCJ gene driven AML and for guiding the prevantion and treatment of AML.

Early and midterm results of thoracic endovascular aortic repair of chronic type B aortic dissection.

OBJECTIVE: The optimal treatment for chronic type B dissection remains controversial. This study reports early and midterm results of thoracic endovascular aortic repair for chronic type B aortic dissection. METHODS: From June 2001 to September 2007, a total of 84 patients with chronic type B aortic dissection underwent thoracic endovascular aortic repair. The time between onset of dissection and thoracic endovascular aortic repair was 13.9 +/- 22.0 months (range, 1-120 months). All patients were followed up from 6 to 86 months (mean, 33.2 +/- 19.2 months). RESULTS: The entry tear was completely sealed in 77 cases (91.7%) during thoracic endovascular aortic repair. The incidence of incomplete seal was 8.3%. The 1-month mortality was 1.2%. One patient had retrograde type A dissection 1 month after the operation. Four patients underwent a second thoracic endovascular aortic repair during follow-up, for endoleak in 3 patients and for newly formed intimal tear in 1 patient. Seven patients (8.3%) died during follow-up. Three died of rupture of the thoracic aorta because of endoleak. The Kaplan-Meier actuarial survival curve showed a 5-year survival of 84.4%. At 5 years, 75.2% of patients were alive with neither endoleak nor reintervention. CONCLUSIONS: Early and midterm results show that thoracic endovascular aortic repair was effective in the treatment of chronic type B aortic dissection. Endoleak was the main cause of death during follow-up. With increased surgical experience and refinement of the stent graft, results are likely to improve in the future.

A study of aortic dimension in type B aortic dissection.

Difference between arch diameter and true lumen diameter in the descending aorta was studied in patients with type B aortic dissection. The diameters of the aortic arch (Proximal ) and mid-descending aorta (Distal ) were measured on computer tomography angiography (CTA) in 20 healthy adults. Forty-two patients with type B aortic dissection who underwent endovascular repair were divided into two groups: an acute group (23 patients) and a chronic group (19 patients). The diameters of the arch (Proximal ) and the true lumen of the mid-descending aorta (Distal ) were measured on digital subtraction angiography (DSA) and CTA. The taper ratio was defined as (Proximal -Distal )/(Proximal )x100%. In the control group, the taper ratio was 13.0+/-4.7% on CTA. In the acute patients group, the taper ratio was 23.6+/-11.3% on DSA and 21.9+/-12.1% on CTA. In the chronic patients group, the taper ratio was 31.5+/-13.6% on DSA and 30.1+/-11.4% on CTA. In both acute and chronic type B aortic dissection, the aorta tapers significantly from arch to true lumen in the descending aorta. Stent-graft with tapered design may be a viable treatment option for endovascular repair of type B aortic dissection.

Endovascular repair of acute type B aortic dissection: early and mid-term results.

OBJECTIVE: To evaluate early and mid-term results of endovascular repair of acute type B aortic dissection by stent graft. METHODS: From June 2001 to May 2005, 63 patients with acute type B aortic dissection underwent stent graft implantation. The study included 59 men and 4 women with an average age of 50.4 +/- 11.4 years (range, 31-80 years). Four patients underwent stent-graft implantation in the acute phase. Fifty-nine patients with acute type B dissection underwent stent-graft implantation 2 weeks after the onset of dissection. All patients were followed up from 1 to 47 months (average, 11.7 +/- 10.6 months). The clinical data of the patients were analyzed. RESULTS: The primary tear was incompletely sealed in three cases. The incidence of incomplete seal was 4.8%. Ascending aortic dissection occurred in three cases. One occurred during operation. The other two occurred at 1 day and 10 months after stent-graft implantation. Two patients died within 30 days after operation. One died of rupture of the ascending aortic dissection. The other one died of acute renal failure. The 1-month mortality was 3.2%. Four patients underwent a second stent-graft implantation before discharge. One year after stent-graft implantation, complete thrombosis of the false lumen in the thoracic aorta was achieved in 98.4% of patients, and the maximum diameter of the descending aorta decreased 11.2% +/- 7.3%. Three patients died within the follow-up time. Mortality during the follow-up period was 4.8%. One patient died of peptic ulcer hemorrhage. Another one died of rupture of the ascending aortic dissection. The third one died of unknown reasons. The actuarial survival curve by the Kaplan-Meier method showed a 4-year survival rate of 89.4%. CONCLUSIONS: Early and mid-term results showed that endovascular repair was effective in treatment of acute type B aortic dissection. With the enrichment of doctors' experience and refinement of the device, better results are expected in the future.

Treating aortic dissection and penetrating aortic ulcer with stent graft: thirty cases.

BACKGROUND: The purpose of this study is to evaluate the feasibility and safety of stent graft in the treatment of aortic dissection and penetrating aortic ulcer. METHODS: From June 2001 to April 2004, 25 patients with aortic dissection and 5 patients with penetrating aortic ulcer received stent-graft implantation. Within this group were 24 male and 6 female patients, with an average age of 52.3 +/- 11.9 years. One patient was diagnosed as type A dissection, 23 patients as chronic type B dissection, 1 patient as acute type B dissection, and the remaining 5 patients with penetrating aortic ulcer. Among the 25 patients with aortic dissection, 3 had contained rupture. All patients were followed up in 1 to 32 months. RESULTS: All patients received stent-graft implantation. There were 5 type I endoleaks. Retrograde ascending aortic dissection occurred during the operation in 1 patient. Two other retrograde ascending aortic dissections occurred in 2 patients 1 day and 7 days, respectively, after stent-graft implantation. Two patients died 1 day and 18 days, respectively, after operation because of rupture of ascending aortic dissection. One-month mortality rate was 6.7%. No death occurred during follow-up time. One patient received a second stent-graft implantation 20 months after the first procedure. CONCLUSIONS: Serious complications may develop after stent-graft implantation, but the early results of treatment of aortic dissection and penetrating aortic ulcer with stent graft were satisfactory. Long-term follow-up was needed.

[Interventional treatment of thoracic aortic dissection with stent-graft placement guided by transesophageal echocardiography].

OBJECTIVE: To evaluate the effect and safety of transfemoral stent-graft placement in combination with transesophageal echocardiography (TEE) in treating DeBakey IIIb aortic dissecting aneurysm. METHODS: Eight male patients [mean age (55.8 +/- 10.2) years, range 28-71 years] with DeBakey IIIb aortic dissection were diagnosed and confirmed by magnetic resonance angiography (MRA). With TEE and fluoroscopy monitoring, transfemoral stent-grafts placement was performed with resulting close of the proximal or both proximal and distal ruptures of aortic dissection. The immediate and follow-up results were assessed. RESULTS: Eleven stent-grafts were placed successfully in the 8 patients, the mean diameter of stents was 29.4 (22-38) mm. Three patients had a mild endoleak and 2 patients had a tiny shunt immediately after operation. Ascending aortic dissection occurred in 1 patient after balloon dilation. The mean stay in hospital was 9 days. All the eight patients were followed up and the mean time of follow-up was 10.5 (6-18) months, 4 residual shunts disappeared. No stents displacement or new endoleak occurred. CONCLUSIONS: Transfemoral stent-graft placement in combination with TEE for the treatment of DeBakey IIIb aortic dissection is a promising method, which is safe, effective and with little injury. Its recent and middle term result is satisfactory, but further observation is needed for long term result.