Diversity of transactivation regions of DMRT1 in vertebrates.

BACKGROUND: Doublesex and mab-3 related transcription factor (DMRT) 1, commonly found in all vertebrates, regulates the transcription of genes involved in the masculinization and maintenance of gonadal somatic cells and/or germline cell development. DMRT1 has a DNA-binding domain called the DM domain and a transcription regulatory region. Unlike the former, there is little knowledge about the latter transcription regulatory region. This study aimed to identify the transcription activation regions of DMRT1 from four species: humans and mice (mammals), leopard geckos (reptiles), and medaka (teleost fish), adding perspectives on evolutionary conservation and diversity. METHODS AND RESULTS: For each species, several expression plasmids of deletion mutants were constructed, and the resultant plasmid and a DMRT1-driven luciferase reporter were co-transfected into cultured cells to measure transactivation ability. The key point of this analysis is that the transactivation ability was normalized by quantifying the expression levels of DMRT1 variants using the HiBiT tag. As a result, two to three transactivation regions were suggested to exist in the C-terminal region of the DM domain in all four species. Among seven regions in DMRT1, the fourth region from the N-terminus contributed to transactivation common to the four species, and the sixth and seventh regions on the C-terminal side differed depending on the species. CONCLUSIONS: These findings indicated that the regions involved in the transactivation ability of DMRT1 could subtly change during evolution, indicating diversity in transactivation domains.

Correlation Between Subgenome-biased DNA Loss and DNA Transposon Activation Following Hybridization in the Allotetraploid Xenopus Frogs.

In certain tetraploid species resulting from interspecific hybridization, one parent's subgenome is known to selectively undergo DNA loss. The molecular mechanisms behind this remain unclear. In our study, we compared the genomes of a standard diploid species with two allotetraploid species from the Xenopus genus, both possessing L (longer) and S (shorter) homoeologous subgenomes. We observed substantial gene losses and intergenic DNA deletions in both the S and L subgenomes of the tetraploid species. Gene losses were around 1,000 to 3,000 for L and 4,000 to 6,000 for S, with especially prominent losses in the S subgenome. Many of these losses likely occurred shortly after interspecific hybridization in both L/S subgenomes. We also deduced frequent large inversions in the S subgenome. Upon reassessing transposon dynamics using updated genome databases, we reaffirmed heightened DNA transposon activity during the hybridization, as previously reported. We next investigated whether S subgenome-biased DNA loss could be correlated with the activation of DNA transposons following hybridization. Notably, distinct patterns were observed in the dynamics of DNA transposons between the L and S subgenomes. Several DNA transposon subfamilies correlated positively with DNA deletions in the S subgenome and negatively in the L subgenome. Based on these results, we propose a model that, upon and after hybridization between two related diploid Xenopus species, the mixture of their genomes resulted in the derepression of DNA transposons, especially in the S subgenome, leading to selective DNA loss in the S subgenome.

Duodenal tuberculosis with gastric outlet obstruction: a case report of successful diagnosis and treatment, with review of literature.

BACKGROUND: Duodenal tuberculosis (TB) is extremely rare, and its diagnosis is challenging owing to the lack of specific symptoms and radiological or endoscopic findings. When it leads to gastric outlet obstruction (GOO), diagnosing it accurately and providing appropriate treatment is crucial. However, this is often overlooked. CASE PRESENTATION: A 35-year-old man presented with abdominal pain, fullness, vomiting, and weight loss. Upper gastrointestinal endoscopy and radiography revealed nearly pinpoint stenosis with edematous and reddish mucosa in the D1/D2 portion of the duodenum. Computed tomography (CT) showed the duodenal wall thickening, luminal narrowing, multiple enlarged abdominal lymph nodes, and portal vein stenosis. Conventional mucosal biopsy during endoscopy revealed ulcer scars. We initially suspected stenosis due to peptic ulcers; however, chest CT revealed cavitary lesions in both lung apices, suggesting tuberculosis. Due to the suspicion of duodenal TB and the need to obtain deeper tissue samples, endoscopic ultrasound-guided fine-needle aspiration (EUS-FNA) was performed. The tissue sample showed caseating granulomas with multinucleated giant cells, and acid-fast bacilli were positive by Ziehl-Neelsen staining. The patient was diagnosed with duodenal TB and subsequent GOO. Because the patient had difficulty eating, surgical intervention was prioritized over antitubercular drugs, and laparoscopic gastrojejunostomy was performed. The patient started an oral diet on the 3rd postoperative day and began antitubercular treatment immediately after discharge on the 11th day. During the 6th month of treatment, endoscopic examination revealed residual duodenal stenosis; however, the bypass route functioned well, and the patient remained asymptomatic. CONCLUSIONS: An aggressive biopsy should be performed to diagnose duodenal TB. EUS-FNA has proven to be a useful tool in this regard. Both nutritional improvement and antitubercular treatment were achieved early and reliably by performing laparoscopic gastrojejunostomy for duodenal TB with GOO.

Neofunctionalization of a Noncoding Portion of a DNA Transposon in the Coding Region of the Chimerical Sex-Determining Gene dm-W in Xenopus Frogs.

Most vertebrate sex-determining genes (SDGs) emerge as neofunctionalized genes through duplication and/or mutation of ancestral genes that are involved with sexual differentiation. We previously demonstrated dm-W to be the SDG in the African clawed frog Xenopus laevis and found that a portion of this gene emerged from the masculinization gene dmrt1 after allotetraploidization by interspecific hybridization between two ancestral species around 17-18 Ma. dm-W has four exons consisting of a noncoding exon 1, dmrt1-derived exons 2 and 3, and an orphan exon 4 (Ex4) of unknown origin that includes coding sequence (CDS). In this study, we searched for the origin of Ex4 and investigated the function of the CDS of this exon. We found that the Ex4-CDS is derived from a noncoding portion of the hAT-10 family of DNA transposon. Evolutionary analysis of transposons and determination of the Ex4 sequences from three other species indicated that Ex4 was generated before the diversification of most or all extant allotetraploid species in subgenus Xenopus, during which time we hypothesize that transposase activity of this hAT superfamily was active. Using DNA-protein binding and transfection assays, we further demonstrate that the Ex4-encoded amino acid sequence increases the DNA-binding ability and transrepression activity of DM-W. These findings suggest that the conversion of the noncoding transposon sequence to the CDS of dm-W contributed to neofunctionalization of a new chimeric SDG in the ancestor of the allotetraploid Xenopus species, offering new insights into de novo origin and functional evolution of chimerical genes.

Activation of DNA Transposons and Evolution of piRNA Genes Through Interspecific Hybridization in Xenopus Frogs.

Interspecific hybridization between two closely related species sometimes resulted in a new species with allotetraploid genomes. Many clawed frog species belonging to the Xenopus genus have diverged from the allotetraploid ancestor created by the hybridization of two closely related species with the predicted L and S genomes. There are species-specific repeated sequences including transposable elements in each genome of organisms that reproduce sexually. To understand what happened on and after the hybridization of the two distinct systems consisting of repeated sequences and their corresponding piRNAs, we isolated small RNAs from ovaries and testes of three Xenopus species consisting of allotetraploid X. laevis and X. borealis and diploid X. tropicalis as controls. After a comprehensive sequencing and selection of piRNAs, comparison of their sequences showed that most piRNA sequences were different between the ovaries and testes in all three species. We compared piRNA and genome sequences and specified gene clusters for piRNA expression in each genome. The synteny and homology analyses showed many distinct piRNA clusters among the three species and even between the two L and/or S subgenomes, indicating that most clusters of the two allotetraploid species changed after hybridization. Moreover, evolutionary analysis showed that DNA transposons including Kolobok superfamily might get activated just after hybridization and then gradually inactivated. These findings suggest that some DNA transposons and their piRNAs might greatly influence allotetraploid genome evolution after hybridization.

A new method to optimize resection area using a radiation treatment planning system and deformable image registration for breast-conserving surgery after neoadjuvant chemotherapy.

BACKGROUND: We devised a breast-conserving surgery (BCS) utilizing a new image-processing and projection technique using a radiation treatment planning system (RTPS) and deformable image registration (DIR) for patients with breast cancer after neoadjuvant chemotherapy (NAC). RTPSs and DIR are commonly used in planning radiation treatment. The purpose of this pilot study was to evaluate the feasibility of our procedure. PATIENTS AND METHODS: Twenty-six patients diagnosed with breast cancer underwent NAC and BCS between November 2014 and May 2020. Multidetector-row computed tomography was performed in the same position used for surgery before and after NAC. In the DIR, CT before NAC was fused to CT after NAC. The RTPS simulated the design of tumor excision, and excision area was projected onto the breast skin utilizing an irradiation device. RESULTS: In 26 patients with breast cancer after NAC, BCS was performed using the processing and projection technique of the RTPS with DIR. Only 1 of 26 patients showed carcinoma present in the surgical margins, and subsequently developed ipsilateral breast tumor recurrence. Mean excised volume was 33.5 cm3 (range, 12.8-62.8 cm3), and percent breast volume excised was 6.8% (range, 2.5-15.7%). CONCLUSIONS: This pilot study confirmed the simplicity and utility of our procedure for minimally invasive BCS in patients with breast cancer after NAC. We will keep evaluating the safety and efficacy of our procedure in more patients.

[Inflammatory Breast Cancer Treated with Multidisciplinary Treatment-A Report of Two Cases].

Case 1: A 77-year-old woman presented with redness and swelling with a peau d'orange appearance in the whole left breast. She was diagnosed with inflammatory breast cancer(T4dN1M0, stage ⅢB of the basal-like subtype). Four courses of FEC followed by 4 courses of docetaxel as the primary systemic therapy were effective. She underwent mastectomy and axillary dissection, and pathological examination revealed a partial response. She received radiation therapy after surgery. At present, 5 years after surgery, the patient is alive without recurrence. Case 2: A 46-year-old woman presented with redness and swelling with a peau d'orange appearance in the whole left breast and edema of the left arm. She was diagnosed with inflammatory breast cancer(T4dN3M1, stage Ⅳ of the HER2 subtype)with bone metastasis and cancerous pleurisy. After 4 courses of FEC as the primary systemic therapy, the breast tumor disappeared, but pleural effusion persisted. Subsequent chemotherapy with pertuzumab, trastuzumab, and docetaxel was effective, as computed tomography showed no lesions. She underwent mastectomy and axillary dissection, and pathological examination revealed a complete response. She received radiation therapy after surgery. At present, 2 years after surgery, the patient is alive without recurrence.

[Study of Phyllodes Tumor of the Breast in 44 Cases of Resection].

Phyllodes tumor is a rare interstitial and epithelial mixed tumor which constitutes 0.3-0.9% of all breast tumors. It is classified into benign, borderline malignant, and malignant. Differentiating this from fibroadenoma is difficult, however, when diagnosed, complete surgical resection is recommended. Patients and results: Forty-four patients who underwent surgery between April 2009 and April 2020 and were diagnosed with phyllodes tumors and the median observation period was 44 months. The average tumor diameter was 39.2 mm, and the surgical procedures performed were mastectomy in 3 cases, partial resection in 26 cases, and extirpation in 15 cases. The surgical margin was positive in 6 cases. Intra-breast recurrence occurred in 4 cases, of which the initial surgery in 3 cases was extirpation. The pathological diagnosis was malignant in 4 cases, malignant in 15 cases, and benign in 25 cases. Conclusion: In this study, intra-breast recurrence occurred in many cases treated by extirpation, and it was considered that the surgical technique for complete resection such as partial resection should be selected.

[A Case Report on Laparoscopic Duodenal Sleeve Resection of GIST at a Distal Portion from the Papilla Vater].

A 21-year-old woman was admitted for preshock due to severe anemia. A 5 cm gastrointestinal stromal tumor(GIST)at the jejunal flexure of her duodenum was diagnosed by enhanced CT examination. We performed a total laparoscopic pancreas- preserving duodenal sleeve resection with a 2 cm margin from the tumor. Functional end-to-end anastomosis was done with the patient lying in a right half lateral decubitus position in order to shift the weight of the tumor and duodenal mesentery to the right to prevent surgical capsule damage. We experienced one case(5.5%)of peritoneal(recurrent)GIST after laparoscopic gastrectomy. However, this is generally a safe and useful procedure for laparoscopic duodenal sleeve resection of duodenal GIST at a distal portion from the papilla Vater, when performed by a skilled team.

[Laparoscopic Hepatectomy of Metastases from Breast Cancer-A Case Report].

A 53-year-old woman, who had been operated with Bt plus Ax plus Ic for right breast cancer 2 years before, revealed an isolated hepatic metastasis at the S4 edge on follow-up computed tomography. The surgical approach is not recommended for distant metastases from breast cancer because of the lack of survival benefits according to the breast cancer guidelines. Nevertheless, the operation could be performed safely and easily for this patient owing to the location and size of the tumor. In addition, the case was a good indication for laparoscopic hepatectomy. We could perform the operation safely with a laparoscope, and the patient was discharged from the hospital on postoperative day 6. Hepatectomy is controversial for metastases from breast cancer. However, in select cases, hepatectomy with a laparoscope can be the therapeutic option as it can improve the survival of patients with hepatic metastases from breast cancer.

[A Case of Gastric Neuroendocrine Carcinoma with Rapidly Increasing Liver Metastasis].

The patient was a 72-year-old man. A gastrointestinal endoscopy was performed because of his anemia and revealed semicircular type 3 tumor in the lower body of the stomach. Biopsy was performed to diagnose adenocarcinoma. Computed tomography(CT)showed no distant metastasis, and the operation was performed 3 weeks after first visit. During operation multiple liver metastasis were recognized, and we performed distal gastrectomy and partial liver resection. The final diagnosis was neuroendocrine carcinoma(NEC). EOB-MRI after surgery revealed multiple liver metastases of 10-20 mm in size, similar to the intraoperative findings, and chemotherapy with cisplatin(CDDP)plus S-1 was started. A significant reduction was recognized after 3 courses of chemotherapy. Gastric NEC has a high proliferative capacity, and distant metastasis may become apparent in a short period of time. Although there is no clear guideline for chemotherapy, CDDP plus S-1 could be 1 treatment option.

[Intraoperative Pancreatoscopy to Assess the Resection Margin in Intraductal Papillary Mucinous Neoplasm-A Case Report].

Intraductal papillary mucinous neoplasm(IPMN)of the pancreas often presents with multifocal lesions. Complete resection without residual skip lesions is essential for complete eradication of the disease. We experienced a case of IPMC in which intraoperative pancreatoscopy was used to determine the surgical margin. Intraoperative pancreatoscopy is a useful and easy method to evaluate the remnant duct and exclude residual tumor. A cystic lesion was incidentally detected in the pancreatic head of a 78-year-old man. Ultrasonography, abdominal computed tomography, magnetic resonance imaging, and endoscopic ultrasound revealed enhancing mural nodules in the pancreatic head and dilation of the entire main pancreatic duct. We performed pancreaticoduodenectomy for mixed IPMN. Intraoperative pancreatoscopy, which was performed to rule out skip lesions, showed no mucosal abnormalities in the remnant duct. The pathological diagnosis was non-invasive intraductal papillary-mucinous carcinoma(IPMC). No signs of recurrence were seen for 6 postoperative months.

[A New Projection-Marking Technique for Breast-Conserving Surgery Using a Radiation Treatment Planning System and Multi-Leaf Collimator in Patients with Breast Cancer-A Pilot Study].

In this study, a breast-conserving surgery(BCS)was performed involving the new image projection technique, using a radiation treatment planning system(RTPS)and a multi-leaf collimator(MLC), for patients with ductal carcinoma in situ (DCIS)and invasive ductal carcinoma(IDC)with ductal components. This study aimed to evaluate the feasibility of this procedure as a pilot study. From June 2014 to May 2017, 27 patients diagnosed with DCIS and IDC with ductal components underwent BCS. In the RTPS, the design of tumor resection was simulated, and the extent of resection was projected to the breast surface using the MLC. BCS was performed using this skin marking. Among 27 patients, 4(14.8%)had carcinoma in the surgical margins. As a pilot study, the simplicity and usefulness of this procedure was confirmed in BCS for patients with DCIS and IDC with ductal components. Further evaluation of its safety and efficacy will be conducted in more patients.

[A Case of Gastrointestinal Stromal Tumor Re-Resected Ten Years after the Resection of Its Recurrence].

The patient was a 75-year-old man who was diagnosed as having jejunal gastrointestinal stromal tumor(GIST)and underwent partial resection of the small intestine including the tumor 12 years ago. Two years after the first resection, recurrence was detected, and a second resection was performed. Ten years after the second resection following recurrence, he took imatinib, and computed tomography(CT)revealed abdominal and liver tumors. We was diagnosed as having GIST recurrence, and a third resection, which included an abdominal tumor resection and partial hepatectomy, was performed. The pathological findings were metastatic abdominal GIST and angiomyolipoma of the liver. The Japanese Clinical Practice Guidelines for GIST suggest a surgical indication only for local recurrence of GIST and resectable liver metastases. Recurrent GIST cannot be completely cured with antineoplastic drugs alone. Multidisciplinary treatment is necessary for long-term survival of patients with recurrent GIST.

[A Case of Transverse Colon Cancer with Liver Metastasis and Tumor Thrombosis of Portal Vein Effectively Treated with Chemotherapy].

The patient was a 70-year-old man. He was diagnosed with advanced transverse colon cancer. A computed tomography (CT)revealed liver metastasis and tumor thrombosis of portal vein. We started combination chemotherapy with capecita- bine/oxaliplatin(CapeOX). Perforation of the tumor was observed 5 days after CapeOX therapy was started. Treatment with abscess drainage and ileostmy, infection was controlled and general condition was improved. After 9 courses of CapeOX, we changed chemotherapy regimen to irinotecan/tegafur-gimeracil-oteracilpotassium (IRIS)due to strong side effects. In CT and FDG-PET examination after 8 courses of IRIS, the tumor of transverse colon, liver metastasis, and the tumor thrombosis of portalvein became unclear. A year and 6 months have passed since chemotherapy was started, recurrence was not observed. For the patients with unresectable colorectal cancer, it is necessary to consider multidisciplinary treatments including chemotherapy while considering the general condition of them.

[Four Cases of Occult Breast Cancer Treated with Breast Conserving Therapy after Neoadjuvant Chemotherapy].

We report 4 patients with occult breast cancer(OBC)who underwent breast conserving therapy(BCT)after neoadjuvant chemotherapy(NAC). All patients complained of axillary tumor and were diagnosed by core needle biopsy. Pathological examination of the axillary lymph nodes proved that 3 cases were adenocarcinomas and 1 case was squamous cell carcinoma, but imaging studies could not depict any primary lesions in the breast and other organs. Since distant metastasis was not observed, BCT with axillary lymph node dissection(ALND)was performed after NAC. All patients were undergoing whole breast irradiation. There were no residual cancer cells in the axillary lymph nodes in case 1 due to the treatment effect of NAC, but lymph node metastasis remained in other 3 cases. In the second case, she followed a rapid outcome, and pulmonary metastasis appeared in 7 months after surgery and died in 10 months. In the other 3 cases, there has not been found local and distant recurrence. Although OBC requires appropriate systemic treatment, present observation indicated that ALND with breast irradiation after NAC could be a useful option.

[A Case Report of Long-Term Survival after Excision for Needle Tract Implantation of Biopsy and Hepatectomy for Cholangiocarcinoma with Gall Bladder Hemorrhage].

A 77-year-old men with abdominal pain suffered from gall bladder hemorrhage and liver abcess was admitted for intensive care of severe acute cholangitis. He had hemobilia which was treated by the endoscopic retrograde bile duct drainage. After 2 months treatment with rest for purulent myelitis, he had neoplastic change of liver abcess diagnosed as intrahepatic bile duct cancer by percutaneous core needle biopsy. He underwent surgery as central bisegmentectomy and left the hospital at home 3 months later after rehabilitation for surgical site infection. He noticed growing mass on right chest 6 months later after hepatectomy and was diagnosed as needle tract implantation in chest wall by CT examination. Pathological exam revealed cholangiocarcinoma by radical excision. He have disease free for 5 years after second operation. Although percutaneous biopsy for hepatic tumor increases risks of needle tract implantation and dissemination, there is an advantage of radical resection for tract implantaion.

[A Case of Repeat Liver Resection after Laparoscopic Resection of a Synchronous Liver Metastasis of Colon Cancer].

A 44-year-old woman with subileus was diagnosed with advanced sigmoid colon cancer with a synchronous liver metasta- sis (segmanet 5/8). Laparoscopic anterior resection was performed, and histological diagnosis was sigmoid colon cancer, 55×40 mm, type 2, tub2>por2, pT3, ly2, v2, pN1, M1a, Stage Ⅳ (Japanese Classification of Colorectal Carcinoma, Eighth edition). Four courses of neoadjuvant chemotherapy (FOLFIRI plus panitumumab) shrank the liver metastasis. Laparoscopic partial liver resection was performed for 285 minutes, with 350 g of blood loss. The patient was discharged 9 days after the operation. Two courses of oral adjuvant chemotherapy (S-1) was performed but discontinued owing to side effects. Seven months after the surgery, computed tomography revealed 2 small liver metastasis (segment 8). Although the sizes were 35 and 5 mm, respectively, the larger mass was closed to the middle and right hepatic vein. Therefore, open hepatectomy was performed for 285 minutes, with 525 g of blood loss. The patient was discharged 9 days after the operation without complication. The patient had no recurrence for 1 year after the last surgery.

Intrahepatic cholangiojejunostomy (Longmire procedure) for recurrent bilioenteric anastomotic stricture with hepatolithiasis.

Interventional procedure via percutaneous transhepatic route is often performed, as an initial treatment, in patients with benign bilioenteric anastomotic stricture. However, surgical management is required in most cases in which radiological intervention is unsuccessful. In this report, we describe a case of a 67-year-old woman with recurrent bilioenteric anastomotic stricture, accompanying bilateral hepatolitiasis after several times of transhepatic interventions. The patient underwent intrahepatic cholangiojejunostomy (Longmire procedure) and cholangioscopic lithotomy after resection of an atrophic left lateral segment resulting from hepatolithiasis. Although the damaged hilar bile duct had to be isolated and divided from the corresponding vasculature for re-anastomosis, it was quite impossible due to severe inflammatory change at the hepatic hilus. We, therefore, anastomosed the intact biliary stump on the cut surface of the left lateral segment to the jejunal loop with a stent tube. The patient's postoperative course was uneventful and she exhibited no evidence of cholangitis during follow-up period of 1 year after surgery. At present, the indications for intrahepatic cholangiojejunostomy for biliary obstruction, are quite limited, but biliary surgeons should keep this procedure in mind at the time of biliary reconstruction for benign proximal bile duct stricture, particularly in cases of multiply operated hilum.

[A successful treatment of conversion chemotherapy by mFOLFOX6 plus cetuximab for initially unresectable synchronous colorectal liver metastases].

A 63-year-old woman with a synchronous huge colorectal liver metastasis was referred to our institution.The lesion was technically diagnosed unresectable because the estimated future remnant liver volume was insufficient due to the invasion of the three hepatic veins and hepatic hilum.She underwent 7 courses of mFOLFOX6 and 14 administrations of cetuximab as conversion chemotherapy.Periodic abdominal CT scans revealed the tumor becoming PR, and she was free of cancer invasion to the left hepatic vein.After the remainder of chemotherapy lasting 4 weeks, right trisectionectomy and combined partial resection of the inferior vena cava and primary closure was performed.The postoperative course was uneventful and the patient was discharged at 20 days after the operation.She underwent chemotherapy postoperatively, and then underwent laparoscopic sigmoidectomy.A conversion chemotherapy using cetuximab may contribute to ward rapidly reducing tumor size and improving the resectability of initially unresectable huge colorectal liver metastases, thus leading to prolonged survival.