[RHPN2 is highly expressed in osteosarcoma cells to promote cell proliferation and migration and inhibit apoptosis].

OBJECTIVE: To screen for aberrantly expressed genes in osteosarcoma cells and investigate the role of RHPN2 in regulating the proliferation, apoptosis, migration and tumorigenic abilities of osteosarcoma cells. METHODS: We used GEO2R to analyze the differential gene expression profile between osteosarcoma cells and normal cells in the GSE70414 dataset. RTqPCR and Western blotting were performed to detect RHPN2 expression in osteosarcoma cell lines MG-63, 143B and SAOS2. Two RHPN2-shRNA and a control NC-shRNA were designed to silence the expression of RHPN2 in 143B cells, and CCK8 assay, colony-forming assay, annexin V-FITC/PI staining and scratch assays were carried out to examine the changes in proliferation, apoptosis and migration of the cells. We also established nude mouse models bearing osteosarcoma xenografts derived 143B cells and RHPN2-shRNA-transfected 143B cells, and assessed the effect of RHPN2 silencing on osteosarcoma cell tumorigenesis using HE staining. Kaplan-Meier survival curves were used to analyze the correlation between RHPN2 expression and survival outcomes of patients with osteosarcoma. RESULTS: RHPN2 expression was significantly upregulated in osteosarcoma cell lines MG-63, 143B and SAOS2 (P < 0.01). Silencing of RHPN2 significantly inhibited the proliferation and migration of 143B cells in vitro, promoted cell apoptosis (P < 0.01), and suppressed tumorigenic capacity of the cells in nude mice. A high expression of RHPN2 was significantly correlated with a poor prognosis of patients with osteosarcoma (P < 0.05). CONCLUSION: RHPN2 is highly expressed in osteosarcoma cells to promote cell proliferation and migration and inhibits cell apoptosis. A high expression of RHPN2 is associated with a poorer prognosis of the patients with osteosarcoma.

Can physiological stimulation prior to ileostomy closure reduce postoperative ileus? A prospective multicenter pilot study.

BACKGROUND: The aim of this study was to assess the impact of ileostomy closure following preoperative physiological stimulation (PPS) on postoperative ileus (POI) in patients with loop ileostomy after low anterior resection for rectal cancer. METHODS: Patients who underwent ileostomy closure between January 2017 and February 2020 in two tertiary referral centers were prospectively included. PPS stimulation was compared to standard treatment. Stimulation was carried out daily during the 15 days prior to ileostomy closure by the patient's self-instillation of 200 ml of fecal contents from the ileostomy bag via the efferent loop, using a rectal catheter. Standard treatment (ST) consisted of observation. Outcomes measures were POI, morbidity, stimulation feasibility, and predictors to ileus. RESULTS: A total of 58 patients were included [42 males and 16 females, median age 67 (43-85) years]. PPS was used in 24 patients, who completed the entire stimulation process, and ST in 34 patients. No differences in preoperative factors were found between the two groups. POI was significantly lower in the PPS group (4.2%) vs the ST group (32.4%); p < 0.01, OR: 0.05 (CI 95% 0.01-0.65). The PPS group had a shorter time to restoration of bowel function (1 day vs 3 days) p = 0.02 and a shorter time to tolerance of liquids (1 day vs 2 days), p = 0.04. Age (p = 0.01), open approach at index surgery, p = 0.03, adjuvant capecitabine (p = 0.01). and previous abdominal surgeries (p = 0.02) were associated with POI in the multivariate analysis. C-reactive-protein values on the 3rd (p = 0.02) and 5th (p < 0.01) postoperative day were also associated with POI. CONCLUSIONS: PPS for patients who underwent ileostomy closure after low anterior resection for rectal cancer is feasible and might reduce POI.

Risk factors for anastomotic leakage following total or subtotal colectomy

Introduction: A higher rate of anastomotic leakage (AL) is reported after ileosigmoid anastomosis (ISA) or ileorectal anastomosis (IRA) in total or subtotal colectomy (TSC) compared with colonic or colorectal anastomosis. Themain aimof the present studywas to assess potential risk factors for AL after ISA or IRA and to investigate determinants of morbidity. Methods: We identified 180 consecutive patients in a prospective referral, single center database, in which 83 of the patients underwent TSC with ISA or IRA. Data regarding the clinical characteristics, surgical treatment, and outcome were assessed to determine their association with the cumulative incidence of AL and surgical morbidity. Results: Ileosigmoid anastomosis was performed in 51 of the patients (61.5%) and IRA in 32 patients (38.6%). The cumulative incidence of ALwas 15.6% (13 of 83 patients). A higher AL rate was found in patients under 50 years-old (p=0.038), in the electivelaparoscopic approach subgroup (p=0.049), and patients in the inflammatory bowel disease (IBD) subgroup (p=0.009). Furthermore, 14 patients (16.9%) had morbidity classified as Clavien-Dindo ≥ IIIA. Discussion: A relatively high incidence of AL after TSC was observed in a relatively safe surgical procedure. Our findings suggest that the risk of AL may be higher in IBD patients. According to our results, identifying risk factors prior to surgerymay improve short-term outcomes. (AU)

Biological mesh reconstruction versus primary closure for preventing perineal morbidity after extralevator abdominoperineal excision: a multicentre retrospective study.

AIM: The aim of the study was to compare the incidence of perineal hernia and the perineal wound morbidity following extralevator abdominoperineal excision (ELAPE) between two groups - primary perineal closure and reconstruction with a biological mesh. METHOD: One hundred and forty-seven consecutive patients who underwent ELAPE for primary rectal cancer between January 2007 and December 2018 in two tertiary referral centres were retrospectively identified from prospective databases. Perineal closure was carried out via primary closure or with a biological mesh (porcine dermal collagen mesh). Outcome measures were perineal hernia and perineal wound morbidity (infection, dehiscence, persistent sinus and chronic pain). RESULTS: A total of 139 patients were included in the study. A prophylactic mesh was used in 80 (57.5%) and primary closure was practised in 59 (42.4%) patients. The median follow-up was 30 (interquartile range 46.88) months. Thirty patients (21.6%) developed perineal hernia. No significant differences were found between prophylactic mesh and primary closure (16.3% vs 23.3%, P = 0.07). The median period between surgery and hernia diagnosis was 8 months in the primary closure group and 24 months in the mesh group (P < 0.01). Perineal wound morbidity was significantly higher in the prophylactic mesh group (55% vs 33.9%, P < 0.01). CONCLUSION: In our study, the use of a biological mesh did not reduce the rate of perineal hernia, although it did delay its appearance. Perineal closure using a biological mesh may increase perineal morbidity, both acute and chronic.

Evidence of epistatic suppression of repeat fruiting in cultivated strawberry.

BACKGROUND: Consumers purchase fresh strawberries all year long. Extending the fruiting season for new strawberry cultivars is a common breeding goal. Understanding the inheritance of repeat fruiting is key to improving breeding efficiency. Several independent research groups using multiple genotypes and analytic approaches have all identified a single genomic region in strawberry associated with repeat fruiting. Markers mapped to this region were used to evaluate breeding parents from the United States Department of Agriculture - Agricultural Research Service (USDA-ARS) strawberry breeding program at Beltsville, Maryland. RESULTS: Markers mapped to repeat fruiting identified once-fruiting genotypes but not repeat-fruiting genotypes. Eleven of twenty-three breeding parents with repeat-fruiting marker profiles were actually once fruiting, indicating at least one additional locus acting epistatically to suppress repeat fruiting. Family segregation ratios could not be predicted reliably by the combined use of parental phenotypes and marker profiles, when using a single-gene model. Expected segregation ratios were calculated for all phenotypic and marker-profile combinations possible from the mapped locus combined with a hypothetical dominant or recessive suppressor locus. Segregation ratios specific to an epistatic suppressor acting on the mapped locus were observed in four families. The segregation ratios for two families were best explained by a dominant suppressor acting on the mapped locus, and, for the other two, by a recessive suppressor. Not all of the observed ratios could be explained by one model or the other, and when multiple families with a common parent were compared, there was no predicted genotype for the common parent that would lead to all of the observed segregation ratios. CONCLUSIONS: Considering all lines of evidence in this study and others, repeat-fruiting in commercial strawberry is controlled primarily by a dominant allele at a single locus, previously mapped by multiple groups. At least two additional genes, one dominant and one recessive, exist that act epistatically to suppress repeat fruiting. Environmental effects and/or incomplete penetrance likely affect phenotype through the suppressor loci, rather than the primary mapped locus. One of the dominant suppressors acts only in the first year, the year the plant is germinated from seed, and not after the plant has experienced a winter.

Anal encirclement with a simple prosthetic sling for faecal incontinence.

BACKGROUND: Millions of people suffer from anal incontinence not currently treated due to the complexity and cost of the surgical techniques in use. The purpose of this report is to evaluate the results of anal encirclement with a simple device in a group of patients with faecal incontinence of multiple causes. METHODS: Surgery was performed as a modified Thiersch procedure through minimal perianal incisions, and a flat band of silicone was introduced to encircle the anus. Results were assessed by clinical response and objectively evaluated by comparing pre- and post-operative manometry, Jorge-Wexner and QOL (Rockwood) scores. RESULTS: Over a period of 5 years, this procedure was performed in 20 women and 13 men, mean age 54 years (27-86 years). Causes of incontinence were iatrogenic (5), obstetric (5), idiopathic (5), restorative proctectomy (5), rectal prolapsed (4), congenital (4), neuropathic (3), trauma (1) and mixed (1). Mortality was nil and morbidity was related to early infection (n = 2) and late skin erosion/infection (n = 2). Early or late breaking of the sling occurred in nine patients, due to the inappropriate method of closing the device. There were 13 explants: 3 as definitive and 10 re-implantations. All but one patient improved the clinical status which was objectively assessed by Jorge-Wexner and QOL scores, with significant differences. CONCLUSIONS: This simple, safe and low-cost technique is useful for treating from disturbing soiling to severe incontinence in all those patients with failed sphincteroplasty or not amenable to sphincter repair. However, a proper device needs to be designed.

Selection of reference genes in Hedysarum coronarium under various stresses and stages of development.

The cultivation of Hedysarum coronarium has generated interest recently for its high yield as a fodder crop, its high protein content, and the presence of condensed tannins in its leaf and stem tissues. Gene expression studies can lead to a better understanding of the biological processes of live organisms. Specifically, reverse transcription followed by quantitative polymerase chain reaction (PCR) represents the most powerful technology for comparing the expression profiles of target genes. The use of reference genes as internal controls to normalize messenger RNA (mRNA) levels is a requirement of quantitative PCR (qPCR). Few studies on reference genes have been performed in plants, and no studies have been performed in H. coronarium. Therefore, the aim of this study was to identify and evaluate reference genes to use in qPCR in H. coronarium. Sulla tissues under two conditions of abiotic stress and at various stages of development were studied to determine adequate reference genes. To optimize the identity and number of reference genes, geNorm and BestKeeper software programs were employed. Based on the results of both analyses, TUA1, TUA2, and UBQ were found to be the most suitable reference genes, and the combination of these three genes was suggested for the accurate normalization of gene expression in sulla tissues.

Selection of housekeeping genes for normalization by real-time RT-PCR: analysis of Or-MYB1 gene expression in Orobanche ramosa development.

Real-time PCR has become the method of choice for accurate and in-depth expression studies of candidate genes. To avoid bias, real-time PCR is referred to one or several internal control genes that should not fluctuate among treatments. A need for reference genes in the parasitic plant Orobanche ramosa has emerged, and the studies in this area have not yet been evaluated. In this study, the genes 18S rRNA, Or-act1, Or-tub1, and Or-ubq1 were compared in terms of expression stability using the BestKeeper software program. Among the four common endogenous control genes, Or-act1 and Or-ubq1 were the most stable in O. ramosa samples. In parallel, a study was carried out studying the expression of the transcription factor Or-MYB1 that seemed to be implicated during preinfection stages. The normalization strategy presented here is a prerequisite to accurate real-time PCR expression profiling that, among other things, opens up the possibility of studying messenger RNA levels of low-copy-number-like transcription factors.

[Hepatic hydatidosis. Radical vs. conservative surgery: 22 years of experience]. / Hidatidosis hepática. Cirugía radical vs. no radical: 22 años de experiencia.

OBJECTIVES: the incidence of hepatic hydatidosis has remarkably decreased in the last years due to the preventive measures adopted to stop the transmission of the parasite. However, surgery carries on being the treatment of choice, although the surgical procedure is still a matter of controversy. The aim of the study was to evaluate the results obtained with the treatment of this condition after two decades according to surgical procedure type. MATERIAL AND METHODS: from 1983 to 2005, 372 patients were operated on for hepatic hydatidic cyst in Hospital Ramón y Cajal. Radical surgery was performed for 162 (43.5%) and conservative surgery for 210 (56.5%). RESULTS: average postoperative hospital stay (8.65 vs. 14.9 days), morbidity (13.3 vs. 31.4%, p < 0.001), and mortality (0 vs. 3.8%, p < 0.01) were lower in the radical surgery group. Recurrence rate was 1.85% after radical surgery versus 11.9% in the conservative surgery group (p < 0.0001). CONCLUSION: radical surgery is associated with lower morbidity, mortality, postoperative hospital stay, and recurrence rates, and represents the treatment of choice for hepatic hydatidosis. However, its indication must depend on the patient characteristics, cyst anatomy, and surgical team experience.

Hidatidosis hepática. Cirugía radical vs. no radical: 22 años de experiencia / No disponible

Introducción: la incidencia de la hidatidosis hepática ha disminuidonotablemente en los últimos años gracias a las medidasde prevención adoptadas para interrumpir la transmisión del parásito.Con todo, la cirugía continúa siendo el tratamiento de elección,si bien su modalidad es todavía motivo de controversia.Objetivos: el objetivo de este trabajo es evaluar los resultadosobtenidos en el tratamiento de esta patología a lo largo de más dedos décadas, atendiendo a la modalidad quirúrgica empleada yafuese cirugía radical o no radical.Material y métodos: se analizaron un total de 372 pacientesintervenidos por quiste hepático hidatídico (QHH) entre 1983-2005 en el Hospital Ramón y Cajal. En162 se efectúa una cirugíaradical (43,5%) y en 210 una no radical (56,5%).Resultados: tanto la estancia media hospitalaria (8,65 días vs.14,9 días) como la morbilidad (13,3 vs. 31,4 %, p < 0,001) y lamortalidad (0 vs. 3,8%, p < 0,01) fueron menores en el grupo decirugía radical. La tasa de recidiva fue del 1,85% tras un abordajeradical frente al 11,9% en los abordajes no radicales (p < 0,0001).Conclusión: la cirugía radical se asocia con una menor morbimortalidad,menor estancia hospitalaria y menor recidiva, constituyendola técnica de elección en la hidatidosis hepática. Sin embargo,su aplicación debe atenerse a las características delpaciente, la anatomía del quiste y el grado de experiencia del equipoquirúrgico

Objectives: the incidence of hepatic hydatidosis has remarkablydecreased in the last years due to the preventive measuresadopted to stop the transmission of the parasite. However, surgerycarries on being the treatment of choice, although the surgical procedureis still a matter of controversy. The aim of the study was toevaluate the results obtained with the treatment of this conditionafter two decades according to surgical procedure type.Material and methods: from 1983 to 2005, 372 patientswere operated on for hepatic hydatidic cyst in Hospital Ramón yCajal. Radical surgery was performed for 162 (43.5%) and conservativesurgery for 210 (56.5%).Results: average postoperative hospital stay (8.65 vs. 14.9days), morbidity (13.3 vs. 31.4%, p < 0.001), and mortality (0 vs.3.8%, p < 0.01) were lower in the radical surgery group. Recurrencerate was 1.85% after radical surgery versus 11.9% in theconservative surgery group (p < 0.0001).Conclusion: radical surgery is associated with lower morbidity,mortality, postoperative hospital stay, and recurrence rates,and represents the treatment of choice for hepatic hydatidosis.However, its indication must depend on the patient characteristics,cyst anatomy, and surgical team experience

First Report of Orobanche crenata on Sulla (Hedysarum coronarium) in Andalusia, Southern Spain.

Crenata broomrape (Orobanche crenata Forsk.), an obligate chlorophyll-lacking root parasite, is the major constraint for growing legume crops in infested soils in southern Spain. Peas (Pisum sativum L), faba beans (Vicia faba L.), and narbon bean (Vicia narbonensis L.) are seriously affected (1,2,4). However, no information is available regarding its ability to attack sulla (Hedysarum coronarium L.), which is an important fodder legume grown in rain-fed conditions, yielding as much as 10,000 kg/ha of dry matter. In 2008, we found broomrape shoots infecting 4 to 5% of sulla plants in a trial field (two land races, P-26 and P-3, in a 500-m2 area) in Cordoba (Andalusia, southern Spain). The shoots developed numerous fertile flowering stalks throughout the field. The previous crop, narbon bean, was heavily infected during 2007 by O. crenata. Infection of sulla plants was confirmed by digging up the plant to verify the attachment of the broomrape plant to the sulla roots. The level of infection was low with only one to two emerged broomrape shoots per sulla plant infested. Morphology was typical of O. crenata. The calyx of the flowers was 13 to 18 mm long with free, bidentate segments. The white corolla of the flowers was 18 to 28 mm long and glandular pubescent. The lips were divergent, large, not ciliate, and filaments inserted 2 to 3 mm above the base of the corolla are hairy at base with glandular hair at the apex. The anthers were brown (3). Specimens were deposited in the Herbarium of the University of Córdoba (identification number COA 45358). To our knowledge, this first report of O. crenata parasitizing sulla shows that control methods for O. crenata will be needed if sulla is to be used as an alternative legume fodder in the infested soils of southern Spain. References: (1) J. I. Cubero and M. T. Moreno. Page 41 in: Some Current Research on Vicia faba in Western Europe. D. A. Bond et al., eds. Luxembourg, 1979. (2) S. Nadal et al. Plant Breed. 126:110, 2007. (3) A. J. Pujadas-Salvá. Page 187 in: Resistance to Orobanche: The State of the Art. Junta de Andalucía, Sevilla, Spain, 1999. (4) D. Rubiales et al. Crop Prot. 22:865, 2003.

Benign rectovaginal fistulas: management and results of a personal series.

BACKGROUND: Treatment of benign rectovaginal fistula has a high failure rate and entails difficult decisions. The purpose of this retrospective study was to clarify the concepts which may improve its management. METHODS: Between 1983 and 2004, 46 consecutive women of median age 41 years were treated by the same surgeon. Etiology of simple fistulas was iatrogenic (n=6), obstetric (n=4) and septic (n=3). Complex fistulas were due to inflammatory bowel diseases (IBD) (n=18, 11 pouchvaginal) or were iatrogenic (n=9), actinic (n=5) or septic (n=1). Surgical techniques included endorectal or vaginal advancement flaps, fistulectomy and sphincteroplasty, vaginal/rectal closure and epiploplasty, restorative proctectomy and restorative proctocolectomy. In 20 patients, a diverting stoma was performed as a single procedure or concomitant to the curative attempt. RESULTS: Overall, 33 of the 39 fistulas (85%) treated for cure healed, including all simple fistulas and 20 complex fistulas (8 iatrogenic, 3 actinic, 2 ulcerative colitis without restorative proctocolectomy; 5 pouch vaginal; 1 septic; 1 Crohn's disease) (p=0.009). The first operation for the fistula was curative in 20 of 39 fistulas, including 10 of 13 simple and 10 of 26 complex fistulas (p=0.023). There was no significant age difference between cured and not-cured patients. CONCLUSIONS: Simple versus complex fistulas is the most determinant factor for healing. In IBD fistulas, ulcerative colitis shows better prognosis than Crohn's disease. For complex fistulas, a temporary diverting stoma seems necessary.

Total anorectal reconstruction: a novel technique.

The aim of this report is to describe a novel technical approach to total anorectal reconstruction after a Miles operation for rectal cancer. Recreation of an internal neosphincter with colonic muscle, the implant of an artificial bowel sphincter (ABS), and the association of a coloplasty constitute a complete substitution of the lost structures and functions. Although the patient developed a late complication which required removal of the ABS, the functional result of the technique can be considered as excellent, to be demonstrated by the incontinence score and quality of life with and without the colostomy.

[Intraductal papillary mucinous tumors of the pancreas: diagnostic and therapeutic approach]. / Neoplasia papilar intraductal mucinosa de páncreas: actitud diagnóstica y terapéutica.

OBJECTIVES: Literature review of the diagnostic and therapeutic approach to intraductal papillary mucinous tumors of the pancreas and report of a new case. CASE REPORT: We present the case of an 85-year-old man treated in our center for steatorrhea and weight loss. Abdominal computerized tomography (CT) showed a heterogeneous pancreatic mass, affecting the hilus, surrounding the superior mesenteric artery and producing dilatation of the biliary tract. Endoscopic retrograde cholangiopancreatography (ERCP) revealed a polypoid neoformation in the lower part of the duodenum and leakage of mucus through the papilla. Biopsy revealed pancreatic adenoma. CONCLUSION: Intraductal papillary mucinous tumor of the pancreas is potentially curable with an uncertain prognosis. Correct diagnosis is required for optimal treatment. One of the greatest problems in this entity continues to be the difficulty of predicting the presence of an infiltrating component, which significantly worsens prognosis. The various diagnostic methods used are abdominal CT, echoendoscopy, ERCP, endoscopy of the Wirsung duct, intraoperative ultrasonography and study of the intraoperative pancreatic resection margin. Treatment requires complete resection of the lesion, although if this provokes pancreatic failure, only tumors with invasive components should be extirpated.

Transient cranial neuropathy in prostatic cancer with bone metastases after rhenium-186-HEDP treatment.

Rhenium-186 (tin) hydroxyethylidene diphosphonate (186Re-HEDP), a bone-seeking radiopharmaceutical, has been successfully used in the treatment of patients with painful bone metastases. Toxicity is usually limited to reversible thrombocytopenia. An infrequent but clinically significant side effect is the occurrence of transient cranial neuropathy. We report on two prostatic cancer patients with metastatic bone cancer. Both patients developed transient cranial neuropathy shortly after treatment with 186Re-HEDP. Transient neuropathy of cranial nerves needs to be distinguished from neurological abnormalities caused by disease progression.

Congenital intrinsic factor deficiency in a Spanish patient.

Vitamin B-12 deficiency was diagnosed in a 26-year-old man. Examinations performed to determine the etiology of the deficiency showed a vitamin B-12 malabsorption in the Schilling test which was corrected by adding intrinsic factor (IF) as well as normal gastric mucosa and acid secretion, although IF in gastric juice was absent. Family study showed normal serum vitamin B-12 levels in the parents, who are first cousins, and siblings. A gastric examination in the father and the sister showed decreased IF secretion, indicating heterozygosity for the disorder.