RESUMO
BACKGROUND rimary hepatic neuroendocrine neoplasms (PHNEN) are exceedingly rare tumors with atypical clinical manifestations, accounting for less than 0.5% of all neuroendocrine tumors. Currently, there is a lack of consensus on their management, and guidelines do not recommend postoperative chemotherapy for patients with stage G1/G2 disease after curative resection. We present a case report of PHNEN, outlining its diagnostic challenges, treatment strategy, and clinical outcomes. CASE REPORT A 31-year-old man presented with jaundice and was initially diagnosed with suspected IgG4-related disease, which initially appeared to respond to steroid therapy, but manifested worsening jaundice 4 months after initial treatment. Subsequent evaluation revealed a PHNEN NET G2 with lymph node metastasis and invasion of the right hepatic artery; and involvement of the hepatic duct at the hepatic hilum, primarily the left hepatic duct. The patient underwent extended left hemi-hepatectomy with caudate lobe resection, bile duct resection, and lymphadenectomy, followed by reconstruction of the right hepatic artery. Postoperatively, the patient received adjuvant chemotherapy consisting of capecitabine (1000 mg bid D1-14) and temozolomide (200 mg qn D10-14) for 6 cycles. Currently, the patient remains disease free 43 months after treatment. CONCLUSIONS PHNEN presents diagnostic challenges due to its rarity and lack of specific markers. Surgical resection remains the cornerstone of treatment, with chemotherapy being considered in select cases with high-risk features. Further research is needed to refine treatment approaches and improve outcomes for patients with PHNEN.
Assuntos
Hepatectomia , Artéria Hepática , Neoplasias Hepáticas , Tumores Neuroendócrinos , Humanos , Masculino , Adulto , Artéria Hepática/cirurgia , Hepatectomia/métodos , Neoplasias Hepáticas/cirurgia , Tumores Neuroendócrinos/cirurgiaRESUMO
Keloids, pathological scars resulting from skin trauma, have traditionally posed significant clinical management challenges due to their persistence and high recurrence rates. Our research elucidates the pivotal roles of lipids and their derivatives in keloid development, driven by underlying mechanisms of abnormal cell proliferation, apoptosis, and extracellular matrix deposition. Key findings suggest that abnormalities in arachidonic acid (AA) synthesis and non-essential fatty acid synthesis are integral to keloid formation. Further, a complex interplay exists between lipid derivatives, notably butyric acid (BA), prostaglandin E2 (PGE2), prostaglandin D2 (PGD2), and the regulation of hyperfibrosis. Additionally, combinations of docosahexaenoic acid (DHA) with BA and 15-deoxy-Δ12,14-Prostaglandin J2 have exhibited pronounced cytotoxic effects. Among sphingolipids, ceramide (Cer) displayed limited pro-apoptotic effects in keloid fibroblasts (KFBs), whereas sphingosine 1-phosphate (S1P) was found to promote keloid hyperfibrosis, with its analogue, FTY720, demonstrating contrasting benefits. Both Vitamin D and hexadecylphosphorylcholine (HePC) showed potential antifibrotic and antiproliferative properties, suggesting their utility in keloid management. While keloids remain a prevalent concern in clinical practice, this study underscores the promising potential of targeting specific lipid molecules for the advancement of keloid therapeutic strategies.
Assuntos
Queloide , Humanos , Queloide/tratamento farmacológico , Queloide/patologia , Matriz Extracelular , Fibrose , Apoptose , Lipídeos/farmacologia , Lipídeos/uso terapêutico , FibroblastosRESUMO
PURPOSE: Nicotinamide adenine dinucleotide (NAD+) is closely related to the pathogenesis of tumors. However, the effect of NAD+ metabolism of gastric cancer (GC) cells on immune cells remains unexplained. We targeted nicotinamide phosphoribosyltransferase (NAMPT), a rate-limiting enzyme in the NAD+ synthesis salvage pathway, to observe its effect in the immune microenvironment. METHODS: NAMPT of GC cell lines was inhibited by using the small molecule inhibitor (FK866) and short hairpin RNA (shRNA). CCK-8 test and flow cytometry were performed to detect cell viability and apoptosis. Immunofluorescence was used to observe changes in mitochondrial membrane potential (MMP).The transfected GC cells (AGS) and patient-derived organoids (PDOs) were cocultured with activated PBMCs, followed by flow cytometric analysis (FCA) for cytokines and inhibitory marker. The level of NAD and ATP of GC cells (AGS & MKN45) was tested combined with NMN and CD39 inhibitor. RESULTS: Targeting NAD+ by FK866 obviously reduced MMP, which ultimately inhibited proliferation and increased the apoptosis of GC cells. NAMPT silencing reduced intracellular NAD and ATPï¼further decreased extracellular adenosine. Meawhile, the cytokines of CD8+T cells were significantly increased after cocultured with transfected AGS, and the expression of PD-1 was distinctly decreased. NMN reversed the effect of shNAMPT and enhanced the immunosuppression. Consistent results were obtained by coculturing PBMCs with PDOs. CONCLUSION: Restraining the function of NAMPT resulted in the functional improvement of effector CD8+ T cells by decreasing extracellular adenosine levels and inducing apoptosis of GC cells simultaneously. Therefore, this study demonstrates that NAMPT can be an effective target for gastric cancer immunotherapy.
Assuntos
NAD , Neoplasias Gástricas , Humanos , NAD/metabolismo , Adenosina/farmacologia , Neoplasias Gástricas/tratamento farmacológico , Linhagem Celular Tumoral , Microambiente Tumoral , Citocinas/metabolismo , Nicotinamida Fosforribosiltransferase/genética , Nicotinamida Fosforribosiltransferase/metabolismo , Trifosfato de Adenosina/metabolismo , Linfócitos T CD8-Positivos/metabolismoRESUMO
OBJECTIVE: To report the safety and efficacy of trans-Douglas Retzius' space-sparing robot-assisted simple prostatectomy (RSS-RASP) in the treatment of large-volume BPH. METHODS: This retrospective study included 24 cases of large-volume (>80 ml) BPH treated by trans-Douglas RSS-RASP from August 2019 to June 2021. The patients ranged in age from 55 to 80 (mean 68.5) years, with an average body mass index of 25.1 (20.5ï¼34.9) kg/m2 , median prostate volume of 132.4 (85.6ï¼235.7) ml, and preoperative tPSA of 10.8 (0.5ï¼37.9) ng/ml, IPSS of 25 (3ï¼35) and quality of life (QOL) score of 5 (3ï¼8). Before surgery, 12 of the patients received catheterization for urinary retention, 1 underwent cystostomy, 2 were complicated with hydronephrosis, 1 had stones and diverticulum in the bladder, and 14 were excluded from the cases of PCa by prostatic biopsy. The operation time, intraoperative blood loss, hemoglobin level on the first day after surgery, blood transfusion, and intra- and postoperative complications were recorded. The patients were followed up for 3 to 21 months postoperatively. Comparisons were made before and after operation in the IPSS, maximum urinary flow rate (Qmax), postvoid residual volume (PVR), QOL score, IIEF score and Male Sexual Health Questionnaire (MSHQ) score. RESULTS: Trans-Douglas RSS-RASP was successfully completed in all the 24 cases, with a mean operation time of 175 (100ï¼285) min, intraoperative blood loss of 200 (50ï¼800) ml, hemoglobin decrease of 25 (4ï¼57) g/L on the first day after surgery, postoperative drainage tube indwelling of 3 (2ï¼7) d, and urinary catheterization of 12 (4ï¼18) d. Six (25%) of the patients received intraoperative blood transfusion, 1 underwent transurethral electrocoagulation hemostasis 1 month after surgery because of postoperative bleeding, and 1 received transurethral resection of the cicatrical adhesive tissue of the bladder neck 12 months after surgery. No other complications occurred postoperatively. The IPSS (3 ï¼»1ï¼7ï¼½), Qmax (19.6 ï¼»9.9ï¼32.1ï¼½ ml/s), PVR (0 ï¼»0ï¼34.9ï¼½ ml) and QOL score (2 ï¼»0ï¼3ï¼½) of the patients were significantly improved after surgery (P < 0.05), but no statistically significant differences were observed in the IIEF (20 ï¼»19ï¼24ï¼½) and MSHQ scores (14 ï¼»13ï¼14ï¼½) as compared with the baseline (P > 0.05). CONCLUSION: Trans-Douglas RSS-RASP is a safe and effective minimally invasive method for the treatment of large-volume (>80 ml) BPH, which can improve the urinary function of the patient after operation.
Assuntos
Hiperplasia Prostática , Robótica , Ressecção Transuretral da Próstata , Humanos , Masculino , Idoso , Próstata/cirurgia , Próstata/patologia , Qualidade de Vida , Hiperplasia Prostática/patologia , Robótica/métodos , Perda Sanguínea Cirúrgica , Estudos Retrospectivos , Hiperplasia/complicações , Hiperplasia/patologia , Ressecção Transuretral da Próstata/métodos , Hemoglobinas , Resultado do Tratamento , Prostatectomia/métodosRESUMO
Organoid culture is a recently developed in vitro three-dimensional (3D) cell culture technology. It has wide applications in tissue engineering studies. However, histological analysis of organoid is quite complex and tedious for researchers. This study proposes a user-friendly, affordable and efficient method for making formalin-fixed paraffin embedded (FFPE) organoid blocks and Optimal Cutting Temperature compound (OCT) embedded frozen organoid blocks. This method implements a key pre-embedding step for preparing paraffin embedded organoid blocks, which could concentrate organoid together without damaging or loss of samples. This method could be used to process even a small number of organoids with high efficiency. In addition, with minor modifications, the method is readily applied for OCT embedded organoid blocks. The slides generated were ready for H&E staining, immunohistochemistry staining and immunofluorescent staining. The method described in this study can be easily used for routine histological analysis of organoid, and could be performed in general pathology labs and requires no dedicated equipment and reagent.
Assuntos
Técnicas Citológicas/métodos , Organoides/citologia , Neoplasias Colorretais/patologia , Fluorescência , Formaldeído , Secções Congeladas , Humanos , Inclusão em Parafina , Fixação de TecidosRESUMO
Immunochromatographic assay (ICA) has been used widely for the onsite monitoring of illegal additives due to its simplicity, speed, and low cost. However, a scanner is commonly required for ICA to achieve quantitative results. In this work, we developed a visual semi-quantitative ICA for sibutramine, a banned additive in diet foods, without the need for a scanner for measurement. Monoclonal antibodies specific for sibutramine were raised and conjugated with upconversion nanoparticles (UCNPs) as the luminescent tracer. ICA was developed by employing multiple test lines to achieve the semi-quantitative detection of sibutramine. Based on the optimal conditions, the cutoff levels (limit of quantitation, LOQ) of T1 line, T2 line, T3 line, and T4 line were 0.02 µg/mL, 0.15 µg/mL, 1.0 µg/mL, and 7.5 µg/mL, respectively, in buffer system. The ICA demonstrated a LOQ at 0.2 mg/kg for sibutramine in diet food samples. The assay (including pretreatment) can be finished within 30 min without the aid of other instruments, except a laser pen. No false positive or false negative results were observed. The results indicated that the proposed method was reliable, simple, and rapid for the screening of sibutramine abuse in diet food samples.
Assuntos
Depressores do Apetite/análise , Cromatografia de Afinidade/métodos , Ciclobutanos/análise , Nanopartículas/química , Animais , Anticorpos Monoclonais/química , Ensaio de Imunoadsorção Enzimática , Contaminação de Alimentos/análise , Limite de Detecção , Camundongos , Espectrometria de FluorescênciaRESUMO
Among several types of ovarian tumors, Sertoli-Leydig cell tumors are considered significantly rare, accounting for less than 1% of all primary ovarian tumors. Hirsutism caused by ovarian tumors accounts for approximately 1% of all cases of hirsutism. We report a case of a woman with a ovarian Sertoli-Leydig cell tumor who presented with hirsutism. A 45-year-old woman (gravida 12, para 2) who experienced menopause when she was 43 years old had excessive hair on her face and lower abdomen since 2 years. Her body mass index was 24.3 kg/m2. She also had hair growth on her upper lip, submandibular area, lower abdomen, vulva, and bilateral tibia (front), and around her breast. She had a Ferriman-Gallwey score of 8. Ultrasound findings revealed a 4.8 × 3.5-cm left adnexal mass. Pelvic computed tomography (CT) findings revealed that her left accessory gland had a low-density mass (CT value, 25 Hu). Her serum testosterone level was 15.80 nmol/l. The patient underwent a laparoscopic left adnexectomy. Subsequently, she was diagnosed with ovarian Sertoli-Leydig cell tumor by immunohistochemical staining. A week after surgery, her serum testosterone level decreased from 15.80 nmol/l to 1.03 nmol/L. Her hirsutism almost completely disappeared 3 months after surgery. It is vitally important to establish the final diagnosis according to the clinical manifestations and laboratory values in addition to imaging studies and laparoscopic examination of a rare coexistence of hirsutism and hyperandrogenemia in a postmenopausal woman based on ovarian pathology.
RESUMO
OBJECTIVES: Intestinal neuronal dysplasia (IND) is a common malformation of the enteric nervous system. Diagnosis requires a full-thickness colonic specimen and an experienced pathologist, emphasizing the need for noninvasive analytical methods. Recently, the methylation level of the Sox10 promoter has been found to be critical for enteric nervous system development. However, whether it can be used for diagnostic purposes in IND is unclear. METHODS: Blood and colon specimens were collected from 32 patients with IND, 60 patients with Hirschsprung disease (HD), and 60 controls. Sox10 promoter methylation in the blood and the Sox10 expression level in the colon were determined, and their correlation was analyzed. The diagnostic efficacy of blood Sox10 promoter methylation was analyzed by receiver operating characteristic curve. RESULTS: The blood level of Sox10 promoter methylation at the 32nd locus was 100% (90%-100%; 95% confidence interval [CI], 92.29%-96.37%) in control, 90% (80%-90%; 95% CI, 82.84%-87.83%) in HD, and 60% (50%-80%; 95% CI, 57.12%-69.76%) in IND specimens. Sox10 promoter methylation in the peripheral blood was negatively correlated with Sox10 expression in the colon, which was low in control, moderate in HD, and high in IND specimens (r = -0.89). The area under the curve of Sox10 promoter methylation in the diagnosis of IND was 0.94 (95% CI, 0.874-1.000, P = 0.000), with a cutoff value of 85% (sensitivity, 90.6%; specificity, 95.0%). By applying a cutoff value of 65%, promoter methylation was more indicative of IND than HD. DISCUSSION: The analysis of Sox10 promoter methylation in the peripheral blood can be used as a noninvasive method for IND diagnosis.
Assuntos
Metilação de DNA , Sistema Nervoso Entérico/anormalidades , Enteropatias/diagnóstico , Doenças do Sistema Nervoso/diagnóstico , Fatores de Transcrição SOXE/genética , Biomarcadores/sangue , Criança , Colo/patologia , Colo/cirurgia , Ilhas de CpG/genética , Diagnóstico Diferencial , Estudos de Viabilidade , Feminino , Doença de Hirschsprung/sangue , Doença de Hirschsprung/diagnóstico , Doença de Hirschsprung/genética , Doença de Hirschsprung/patologia , Humanos , Enteropatias/sangue , Enteropatias/genética , Enteropatias/patologia , Mucosa Intestinal/inervação , Masculino , Doenças do Sistema Nervoso/sangue , Doenças do Sistema Nervoso/genética , Doenças do Sistema Nervoso/patologia , Valor Preditivo dos Testes , Regiões Promotoras Genéticas/genética , Curva ROC , Fatores de Transcrição SOXE/metabolismo , Análise de Sequência de DNARESUMO
OBJECTIVE: To investigate the clinical feasibility and effect of nerve-sparing robot-assisted laparoscopic radical cystectomy (NSRA-LSRC). METHODS: We retrospectively reviewed the clinical data on 12 cases of NSRA-LSRC performed from March 2016 to May 2018. The patients were aged 45 to 65 years old and all potent before surgery, with a mean IIEF-5 score of >17. The surgical procedure involved excision of the bladder and prostate and dissection of the pelvic lymph nodes, with preservation of the bilateral neurovascular bundles, internal accessory pudendal artery and pubic bladder complex. All the patients were advised to take PDE5I postoperatively and followed up for the sexual function with the IIEF-5 scores. RESULTS: Surgical procedures were completed successfully, all with negative surgical margins. Postoperative pathology confirmed invasive high-grade urothelial carcinoma or carcinoma in situ in all the cases, including 11 cases in stage T2N0M0 or below and 1 case in stage T3aN0M0. There were no serious intraoperative or postoperative complications, nor recurrence or metastasis during the follow-up period of 12ï¼36 (20.7 ± 8.0) months. The IIEF-5 scores of the patients at 3, 6 and 12 months after operation were 10.9 ± 6.9, 12.3 ± 6.9 and 14.1 ± 8.0, respectively. At 12 months, satisfactory sexual intercourse was achieved with the help of potency-enhancing medicine in 5 cases (41.7%), penile erection insufficient for sexual intercourse in 3 cases (25%), and no erection in 4 cases (33.3%). CONCLUSIONS: Nerve-sparing robot-assisted laparoscopic radical cystectomy can maximally preserve the sexual function of the patients with urinary bladder carcinoma.
Assuntos
Cistectomia/métodos , Laparoscopia , Procedimentos Cirúrgicos Robóticos , Neoplasias da Bexiga Urinária/cirurgia , Idoso , Humanos , Masculino , Pessoa de Meia-Idade , Recidiva Local de Neoplasia , Tratamentos com Preservação do Órgão , Ereção Peniana , Estudos RetrospectivosRESUMO
A sensitive full-automated micromagnetic particles (MMPs) based competitive chemiluminescent immunoassay (CLIA) was developed to detect cortisol in milk. Polyclonal antibody (pAb) with good specificity against cortisol was produced. The antigen (cortisol-OVA) was labeled with acridinium ester (cortisol-OVA-AE) as signal tracer. During the detection, the free cortisol in sample will compete with cortisol-OVA-AE for binding to pAb. To capture pAb, MMPs conjugated with goat anti-rabbit IgG was added. The whole immunoassay process (exclude sample pretreatment) was performed by automatic chemiluminescence immunoassay instrument, which could consume less test time (within 40â¯min) and avoid error from manual operation. The method showed a good detection limit of 0.12â¯ng/mL, a broad linear range from 0.42 to 72.27â¯ng/mL for cortisol detection, negligible cross-reactivity with related analogues and satisfied recovery (84.3%-102.3%) for spiked milk samples test. Simultaneously, since the results of proposed method had no significant difference with those of LC-MS/MS, the proposed method was confirmed to have a potential applicability for rapidly monitoring cortisol in the food.
Assuntos
Hidrocortisona/análise , Imunoensaio/métodos , Medições Luminescentes/métodos , Leite/química , Animais , Anticorpos/imunologia , Análise de Alimentos/métodos , Contaminação de Alimentos/análise , Cabras , Haptenos/imunologia , Imunoensaio/instrumentação , Limite de Detecção , Medições Luminescentes/instrumentação , Magnetismo , Masculino , Coelhos , Reprodutibilidade dos TestesRESUMO
OBJECTIVE: To investigate the application of Retzius-sparing robot-assisted radical prostatectomy (RS-RARP) in the treatment of early-stage prostate cancer. METHODS: We retrospectively analyzed the clinical data about 10 cases of early-stage prostate cancer treated by RS-RARP with the Da Vinci Robot Surgical System from September to October 2016. RESULTS: All the operations were successfully completed without positive surgical margins. The operation time was 170ï¼250 min (ï¼»196±25ï¼½ min), the intraoperative blood loss was 150ï¼500 ml (ï¼»260±128ï¼½ ml), the postoperative hospital stay was 6ï¼7 days, and the catheterization time was 14 days. Urinary continence occurred after catheter removal in 1 patient and was recovered 1 month later. CONCLUSIONS: RS-RARP is a safe, effective and reliable method for the treatment of prostate cancer and conducive to the early recovery of urinary continence.
Assuntos
Laparoscopia/métodos , Prostatectomia/métodos , Neoplasias da Próstata/cirurgia , Procedimentos Cirúrgicos Robóticos , Perda Sanguínea Cirúrgica , Humanos , Tempo de Internação , Masculino , Margens de Excisão , Pessoa de Meia-Idade , Duração da Cirurgia , Período Pós-Operatório , Neoplasias da Próstata/patologia , Estudos RetrospectivosRESUMO
A simple and fast homogeneous fluorescent polarization immunoassay (FPIA) was developed for the determination of furaltadone and its metabolite 3-amino-5-methylmorpholino-2-oxazolidinone (AMOZ). Monoclonal antibody with high cross-reactivity to furaltadone and the nitrophenyl derivative of AMOZ (NPAMOZ) were produced against a novel immunogen and the effects of several synthesized tracers on FPIA sensitivity studied. The proposed FPIA, using an optimum antibody and tracer pair, had an IC50 of 4.3 µg L⻹ and limit of detection at 0.6 µg L⻹ for furaltadone, and 2.7 µgL⻹ and 0.3 µg L⻹ for NPAMOZ. Recoveries of furaltadone from animal feeds by FPIA ranged from 79.6 to 87.7%, while recoveries of AMOZ from animal tissues ranged from 72.9 to 83.1%. Good correlation (R>0.99) between the results of this FPIA and a standard analytical method was obtained. The FPIA does not require separation or washing steps and the total time required for equilibrium of the antibody-tracer interaction is only 10 min. These results indicated that the proposed FPIA offers great potential and utility for the high throughput screening of furaltadone residues in animal feed and its metabolite AMOZ residues in animal tissues.
Assuntos
Ração Animal , Anticorpos Monoclonais/imunologia , Imunoensaio de Fluorescência por Polarização , Ensaios de Triagem em Larga Escala/métodos , Morfolinos/farmacocinética , Nitrofuranos/análise , Nitrofuranos/metabolismo , Oxazolidinonas/análise , Oxazolidinonas/metabolismo , Animais , Galinhas , Feminino , Peixes , Camundongos , Camundongos Endogâmicos BALB C , Estrutura Molecular , Morfolinos/análise , Morfolinos/metabolismo , Nitrofuranos/química , Nitrofuranos/farmacocinética , Oxazolidinonas/química , Oxazolidinonas/farmacocinética , Suínos , Distribuição TecidualRESUMO
Anorectal malformations (ARMs, congenital obstruction of the anal opening) are among the most common birth defects requiring surgical treatment (2-5/10 000 live-births) and carry significant chronic morbidity. ARMs present either as isolated or as part of the phenotypic spectrum of some chromosomal abnormalities or monogenic syndromes. The etiology is unknown. To assess the genetic contribution to ARMs, we investigated single-nucleotide polymorphisms and copy number variations (CNVs) at genome-wide scale. A total of 363 Han Chinese sporadic ARM patients and 4006 Han Chinese controls were included. Overall, we detected a 1.3-fold significant excess of rare CNVs in patients. Stratification of patients by presence/absence of other congenital anomalies showed that while syndromic ARM patients carried significantly longer rare duplications than controls (P = 0.049), non-syndromic patients were enriched with both rare deletions and duplications when compared with controls (P = 0.00031). Twelve chromosomal aberrations and 114 rare CNVs were observed in patients but not in 868 controls nor 11 943 healthy individuals from the Database of Genomic Variants. Importantly, these aberrations were observed in isolated ARM patients. Gene-based analysis revealed 79 genes interfered by CNVs in patients only. In particular, we identified a de novo DKK4 duplication. DKK4 is a member of the WNT signaling pathway which is involved in the development of the anorectal region. In mice, Wnt disruption results in ARMs. Our data suggest a role for rare CNVs not only in syndromic but also in isolated ARM patients and provide a list of plausible candidate genes for the disorder.
Assuntos
Anus Imperfurado/genética , Anus Imperfurado/fisiopatologia , Variações do Número de Cópias de DNA , Duplicação Gênica , Peptídeos e Proteínas de Sinalização Intercelular/genética , Animais , Malformações Anorretais , Povo Asiático , Aberrações Cromossômicas , Feminino , Dosagem de Genes , Regulação da Expressão Gênica , Estudo de Associação Genômica Ampla , Humanos , Peptídeos e Proteínas de Sinalização Intercelular/metabolismo , Masculino , Camundongos , Camundongos Endogâmicos ICR , Fenótipo , Polimorfismo de Nucleotídeo Único , Estudos Prospectivos , Via de Sinalização WntRESUMO
A heterologous competitive indirect enzyme-linked immunosorbent assay (ciELISA) for the determination of the furaltadone metabolite 3-amino-5-morpholinomethyl-2-oxazolidinone (AMOZ) was developed. AMOZ was derivatised with 2-(4-formylphenoxy) acetic acid or 2-(3-formylphenoxy) acetic acid to obtain two novel immunizing haptens. The ability of these haptens in producing specific polyclonal antibodies against the nitrophenyl derivative of AMOZ (NPAMOZ) was compared with that of traditional immunizing haptens (derivatised AMOZ with 3-carboxybenzaldehyle or 4-carboxybenzaldehyle). The results indicated that the novel immunizing haptens were able to produce antibodies with almost a two-fold improvement in sensitivity of the ciELISA for NPAMOZ in comparison with the existing antibody based ELISAs. The differences in sensitivity were explained by the molecular modeling of the lowest energy conformations of NPAMOZ and the haptens. Another novel hapten, derivatised AMOZ with 2-oxoacetic acid, was synthesized and used as a heterologous coating hapten. The results showed that this strategy of using only a partial structure of the target molecule as the coating hapten was able to obtain a two to three-fold improvement in sensitivity. This study provided a modern approach for the development of an immunoassay with improved sensitivity for the metabolites of nitrofuran antibiotics.
Assuntos
Ensaio de Imunoadsorção Enzimática , Haptenos/química , Morfolinas/análise , Nitrofuranos/metabolismo , Oxazolidinonas/análise , Oxazolidinonas/metabolismo , Animais , Formação de Anticorpos , Especificidade de Anticorpos , Galinhas/metabolismo , Peixes/metabolismo , Haptenos/imunologia , Imunização , Imunoconjugados , Imunoglobulina G/imunologia , Masculino , Modelos Moleculares , Penaeidae/metabolismo , Coelhos , Reprodutibilidade dos Testes , Suínos/metabolismoRESUMO
A monoclonal antibody-based competitive indirect enzyme-linked immunosorbent assay (ELISA) with improved sensitivity and specificity for the determination of furaltadone metabolite 5-methylamorpholino-3-amino-2-oxazolidone (AMOZ) was described. AMOZ was derivatized with 2-(3-formylphenoxy)acetic acid and coupled with bovine serum albumin to form a novel immunogen. BABL/c mice were immunized and monoclonal antibody specific to the nitrophenyl derivative of AMOZ (NP-AMOZ) was produced and characterized. Four other haptens with different heterology to the immunizing hapten were synthesized and coupled to ovalbumin as coating antigens to study the effect of heterologous coating on assay sensitivity. Under the optimized heterologous coating format, the competitive indirect ELISA showed very high sensitivity to NP-AMOZ, with an IC(50) of 0.14 µg/L and limit of detection of 0.01 µg/L. The assay showed high specificity toward NP-AMOZ, and negligible cross-reactivity with analogous compounds was observed. The average recoveries of AMOZ from spiked fish and shrimp samples were estimated to range from 81.0 to 104.0%, with coefficients of variation below 20%. Good correlation was obtained between the results of ELISA analysis and of standard liquid chromatography-tandem mass spectrometry analysis. These results indicated that the proposed ELISA is ideally suited as a monitoring method for AMOZ residues at trace level.
Assuntos
Anticorpos Monoclonais/imunologia , Crustáceos/química , Ensaio de Imunoadsorção Enzimática/métodos , Peixes/metabolismo , Morfolinas/análise , Oxazolidinonas/análise , Animais , Limite de Detecção , Morfolinas/imunologia , Oxazolidinonas/imunologiaRESUMO
Rhabdomyosarcoma (RMS) is one of the most frequent soft tissue sarcomas in children. It is sometimes difficult to distinguish it from other small round cell tumors (SRCTs) depending on microscopic observations; although their treatment and prognosis varied widely, the same happens between alveolar RMS (ARMS) and embryonal RMS (ERMS). The role of PAX3/PAX7-FKHR fusion gene has been reported in ARMS but not in ERMS and SRCT. The aim of this study was to explore its value in RMS diagnosis and differentiation. Ninety-eight patients with ARMS (n = 13), ERMS (n = 25), pleomorphic RMS (n = 5), Ewing sarcoma (n = 11), neuroblastoma (n = 18), lymphoma (n = 24), and uncertain SRCT (n = 2) were analyzed. One hundred fifteen RNA samples were extracted from the primary tumor tissue at initial presentation and relapse. One-step reverse transcriptase polymerase chain reaction assays for the PAX3/PAX7-FKHR fusion transcripts were performed. Molecular findings were compared with original histologic diagnoses. PAX3-FKHR fusion transcript was detected in 9 ARMS samples, PAX7-FKHR fusion transcript was detected in 7 ARMS samples, and 2 uncertain SRCTs were detected; none of them were detected in ERMS, Ewing sarcoma, neuroblastoma, and lymphoma. Direct sequencing of PAX3 coding regions revealed a heterozygous mutation AâG (nt1380) at codon 448 (AATâGAT), resulting in substitution of Asn-448 for Asp. Detection of PAX3/PAX7-FKHR fusion transcripts by 1-step reverse transcriptase polymerase chain reaction is a novel tool for RMS diagnosis and differentiation.
Assuntos
Biomarcadores Tumorais/análise , Proteínas de Fusão Oncogênica/análise , Reação em Cadeia da Polimerase Via Transcriptase Reversa/métodos , Rabdomiossarcoma/diagnóstico , Neoplasias de Tecidos Moles/diagnóstico , Sequência de Bases , Biomarcadores Tumorais/genética , Análise Mutacional de DNA , Humanos , Dados de Sequência Molecular , Rabdomiossarcoma/classificação , Rabdomiossarcoma/genética , Neoplasias de Tecidos Moles/classificação , Neoplasias de Tecidos Moles/genéticaRESUMO
C-type natriuretic peptide (CNP) is a 22-amino acid peptide and act as a local paracrine or autocrine regulator. There is growing evidence that CNP is involved in male reproductive processes. To investigate the role of CNP during spermatogenesis, we measured the mRNA expression of CNP and its specific membrane-bound natriuretic peptide receptor-B (NPR-B) using real-time RT-PCR in the testes of normal rats on different postnatal days. After that spermatogenesis dysfunction model induced by ornidazole was established with the aim to study the correlation of CNP with spermatogenic dysfunction. Then, Sertoli cells from 18- to 22-day-old healthy male rats were cultured in the presence of different CNP concentrations (1×10(-6), 1×10(-7) and 1×10(-8) mol l(-1)), and the mRNA expression levels of androgen-binding protein, inhibin B and transferrin were examined at 0 min, 30 min, 1 h, 2 h, 4 h, 8 h, 12 h, 24 h and 48 h. During the postnatal development of rat testes, the highest mRNA expression levels of CNP and NPR-B were found at postnatal D(0), and the levels then declined gradually, with a second CNP peak at postnatal D(35). In the ornidazole-induced infertile rat testes, CNP gene expression was lower than in the uninduced rats (P<0.05), while NPR-B gene expression was greater (P<0.05). In cultured Sertoli cells, supplementation with CNP stimulated the gene expression of androgen-binding protein/inhibin B/transferrin, particularly at 12 h, and 1×10(-7) mol l(-1) CNP had the highest upregulation effect. The gene expression levels of CNP/NPR-B in rat testes at different postnatal stages and in infertile rat testes indicated that CNP may participate in the physiology and/or pathology related to spermatogenesis. Moreover, CNP regulated endocrine function in Sertoli cells. Taken together, these results showed that CNP is closely tied to spermatogenesis.
Assuntos
Peptídeo Natriurético Tipo C/genética , Peptídeo Natriurético Tipo C/metabolismo , Espermatogênese/genética , Espermatogênese/fisiologia , Testículo/metabolismo , Proteína de Ligação a Androgênios/genética , Animais , Sequência de Bases , Células Cultivadas , Primers do DNA/genética , Regulação da Expressão Gênica no Desenvolvimento , Infertilidade Masculina/induzido quimicamente , Infertilidade Masculina/genética , Infertilidade Masculina/metabolismo , Inibinas/genética , Masculino , Peptídeo Natriurético Encefálico/genética , Peptídeo Natriurético Encefálico/metabolismo , Peptídeo Natriurético Tipo C/farmacologia , Ornidazol/toxicidade , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Ratos , Ratos Sprague-Dawley , Proteínas Recombinantes/farmacologia , Células de Sertoli/efeitos dos fármacos , Células de Sertoli/metabolismo , Testículo/efeitos dos fármacos , Testículo/crescimento & desenvolvimento , Transferrina/genéticaRESUMO
PURPOSE: To assess the efficacy of contrast-enhanced ultrasonography (CEUS) with Sonovue in the evaluation of therapeutic response to radiofrequency ablation (RFA) of renal cell carcinoma (RCC). MATERIALS AND METHODS: In a recent 3 years, 63 patients (mean age, 60 years; range 26-81 years) with 64 RCCs were treated by RFA. The lesions had a diameter between 1.8 and 9.8 cm (average diameter, 3.1 cm). The indications for RFA treatment included chronic renal insufficiency (n = 10), presence of solitary kidney (n =3), bilateral renal carcinoma (BRCC) (n =2), advanced age (n =12), significant medical comorbidity (n =29) or refusal of conventional therapy (n =7). Tumors were treated by laparoscopy-assisted (n =41), open surgical (n =18) or percutaneous US guidance (n =4). Follow-up CEUS and contrast-enhanced CT were performed 1 month after treatment to assess the necrotic area. Technical success was defined as elimination of areas that enhanced at imaging within the entire tumor. RESULTS: On the 1-month CEUS and CT imaging after RFA, 62 of 64 tumors (96.9%) were successfully ablated with one session, and residual tumors were found in two RCCs. One of the two tumors was subjected to additional RFA treatment. We could not obtain a complete ablation in the other tumor of a patient with solitary kidney. The diagnostic concordance between the CEUS and 1-month follow-up CT was 100%. Sixty-one patients survived in the follow-up phase which ranged from 2 to 34 months. One patient with solitary kidney died of systemic disease progression and one patient was lost to follow-up. Of the 61 tumors without residual on both CT and CEUS after RFA, four had suspicious findings of recurrence on follow-up CEUS, and two of them were confirmed by subsequent CT examination. With CT as the reference imaging procedure in the assessment of renal tumor ablation, the sensitivity, specificity, positive predictive value, and negative predictive value of CEUS for detecting recurrence during follow-up were 100%, 96.6%, 50%, and 100%. CONCLUSION: Despite its limitation of false-positive value, CEUS is potentially effective in assessing the therapeutic response to RFA of RCC.
Assuntos
Carcinoma de Células Renais/diagnóstico por imagem , Carcinoma de Células Renais/cirurgia , Ablação por Cateter , Meios de Contraste , Neoplasias Renais/diagnóstico por imagem , Neoplasias Renais/cirurgia , Fosfolipídeos , Hexafluoreto de Enxofre , Adulto , Idoso , Idoso de 80 Anos ou mais , Feminino , Humanos , Masculino , Pessoa de Meia-Idade , Tomografia Computadorizada por Raios X , UltrassonografiaRESUMO
OBJECTIVE: To evaluate the clinical feasibility of radiofrequency ablation-assisted laparoscopic partial nephrectomy (RF-LCPN) without renal hilar vessel clamping. METHODS: From January 2009 to June 2010, 14 cases with renal tumors were treated with RF-LCPN. The mean tumor size was 3.1 cm (range: 2.2 - 4.0 cm). All cases were staged at T(1a)N(0)M(0). The preoperative serum creatinine (SCr) was (87 ± 36) µmol/L and glomerular filtration rate (GFR) (45 ± 11) ml/min. RESULTS: RF-LCPN was technically successful in all patients (14 tumors). The mean operative time was (152 ± 46) min and the mean blood loss (89 ± 52) ml. All tumor margins were negative. One patient with a brief period of urine leakage from lower pole calix was managed successfully by ureteral stenting and Foley catheter drainage of bladder. The postoperative hospital stay was (5 ± 2) days. The postoperative SCr was (90 ± 38) µmol/L and GFR (41 ± 12) ml/min. There was no statistic post-operative change of SCr and GFR (P > 0.05). All patients completed a minimum follow-up of 6 months (mean: 17 months, range: 6 - 25 months). No evidence of local or port-site recurrence or metastasis was found. CONCLUSION: As an accurate and effective intervention with a relatively low incidence of complications, RF-LCPN may eliminate renal warm ischemia. But its long-term efficacy remains to be further studied.
Assuntos
Laparoscopia , Nefrectomia , Constrição , Humanos , Neoplasias Renais , Recidiva Local de NeoplasiaRESUMO
OBJECTIVE: Functional constipation (FC) is a common complaint in childhood but the prevalence still remains unclear, the aim of this study was to explore the prevalence of FC in childhood in the northern parts of China. METHODS: A screening program on FC in childhood was carried out in 5 northern cities (Beijing, Tianjin, Shenyang, Changchun, Harbin) of China according to symptoms under the Rome III criteria. Random clustered sampling of the inhabitants was carried out under stratification of cities, geographic zones, schools or nurseries. Sample size of each area was in proportion to the population of the area. The range of age was 4-14. All subjects under study were requested to fill in a questionnaire distributed by the teachers. The screening program was carried out immediately after a section survey was completed. All together, 20 000 questionnaires were distributed with 19 638 retrievals. According to the result of the screening, a small number of patients who met the criteria were further selected to undergo a detailed clinical examination in the hospital including laboratory examination, colonic transit time, defecography or/and barium enema, electromyologram and anorectal manometry to exclude organic disease of the colon. Simultaneously, a detailed questionnaire was requested to fill under the assistance of trained doctors or medical students. Prevalence of FC of the population was adjusted by the rate of correct diagnosis from the detailed study. RESULTS: The adjusted point prevalence of FC in 5 northern cities of China according to Rome III criteria was 4.73% with higher prevalence rate of FC seen in the Beijing area (5.02%) than in other cities (4.82%, 4.76%, 4.27%, 4.40%, with P < 0.001). Male to female ratio was 1.26:1 with majority of the FC fell in the 4 - 6 year olds (5.76%). CONCLUSION: FC appeared a common disorder in childhood in the northern parts of China which called for greater attention. Higher prevalence was noted in the age groups of 4-6.