Single-Cell Characterization of Malignant Phenotypes and Developmental Trajectories of Adrenal Neuroblastoma.

Neuroblastoma (NB), which is a subtype of neural-crest-derived malignancy, is the most common extracranial solid tumor occurring in childhood. Despite extensive research, the underlying developmental origin of NB remains unclear. Using single-cell RNA sequencing, we generate transcriptomes of adrenal NB from 160,910 cells of 16 patients and transcriptomes of putative developmental cells of origin of NB from 12,103 cells of early human embryos and fetal adrenal glands at relatively late development stages. We find that most adrenal NB tumor cells transcriptionally mirror noradrenergic chromaffin cells. Malignant states also recapitulate the proliferation/differentiation status of chromaffin cells in the process of normal development. Our findings provide insight into developmental trajectories and cellular states underlying human initiation and progression of NB.

Long-term prognosis of low-risk neuroblastoma treated by surgery alone: an experience from a single institution of China.

BACKGROUND: Low-risk neuroblastomas have favorable biologic characteristics. Children Oncology Group (COG) proposed that surgical resection of the primary tumor was sufficient. We evaluated the long-term prognosis of surgery alone for patients with low-risk neuroblastoma in China. METHODS: A total of 34 patients with low-risk neuroblastoma were treated in our center between Jan 2009 and Dec 2013. The medical records of these patients were reviewed. RESULTS: The primary lesion was located in the adrenal gland in 19 patients, the retroperitoneum in 5, the posterior mediastinum in 9 and the neck in 1. The tumor diameters and volumes were 1.80-10.0 cm (average 5.5 ± 2.3 cm) and 1.28-424.10 cm3 (average 58.81 ± 92.00 cm3), respectively. The stages of the patients were as follows: stage I in 25, stage II in 7, and stage IVs in 2. All patients were in the low-risk group according to COG risk stratification criteria. No patients showed MYCN amplification. The primary tumors of all patients were completely resected. Nine adrenal tumors were completely resected by laparoscopy. All patients were successfully followed for 66-115 (average 89.71 ± 16.17) months. Recurrence was observed in 4 patients. In addition to one local recurrence, another three recurrences were metastases. The lesions were effectively controlled in all patients with recurrences. All patients survived, including 28 cases of tumor-free survival; the 4-year overall and event-free survival rates were both 100%. CONCLUSIONS: Surgery alone is a safe and effective treatment strategy for low-risk neuroblastoma. Recurrent lesions may be controlled and treated by rescue chemotherapy and surgery.

Primary gastric tumors in infants and children: 15 cases of 20-year report.

PURPOSE: Primary gastric tumors in infants and children are rare, and their diagnosis and treatment have not been standardized to date. The objective of the present retrospective study was to analyze the clinical characteristics of these tumors and explore possible improvements in their diagnosis and treatment. METHODS: The study included 15 children with a diagnosis of primary gastric tumor confirmed by pathology. Clinical manifestations, diagnostic methods and treatment were analyzed retrospectively, and postoperative conditions were assessed in follow-up evaluations. RESULTS: There were nine boys and six girls aged 8 months to 13 years. The main presenting symptoms were abdominal pain, melena, fever of undetermined origin and pallor. Children were assessed by ultrasound, CT and upper gastrointestinal barium meal or gastroscopy and showed abdominal blockage or polypoid space-occupying lesions. All patients underwent surgery as initial treatment, and four patients received postoperative chemotherapy. During the follow-up period from 3 to 92 months, four cases were lost, one patient died of metastatic disease, two patients showed recurrence, and the remaining patients were alive without recurrence or progression. CONCLUSIONS: Owing to the atypical and often asymptomatic presentation of primary gastric tumors, careful evaluation using imaging modalities is critical in suspicious cases. Most primary gastric tumors in infants and children are benign or borderline. The prognosis, except in gastric carcinoma, is excellent with close follow-up when complete resection is achieved.

Laparoscopic Radical Nephrectomy of Wilms' Tumor and Renal Cancer in Children: Preliminary Experience from a Two-Center Study in China.

BACKGROUND: To report the preliminary experience of laparoscopic radical nephrectomy (LRN) in children with Wilms' tumor (WT) and renal cancer. PATIENTS AND METHODS: From January 2010 to October 2013, the medical records of 7 cases of WT or renal cancer in children treated by LRN at two medical centers in China were reviewed. RESULTS: All the patients were treated by LRN, and 3 of them underwent preoperative chemotherapy before surgery. The biggest tumor size was 10 cm without crossing the lateral edge of the vertebra at the time of operation. The median hospital stay was 8.5 days (range, 6-11 days). The pathologic investigation showed 5 cases of WT, 1 case of rhabdoid tumor, and 1 case of renal cell carcinoma. With a median follow-up of 1.9±1.5 years (range, 0.3-2.9 years), six children were in complete remission, and the remaining one was lost to follow-up. None of these patients presented evidence of oncological complications (tumoral recurrences, port-site implantation, or secondary lung metastases), and no small bowel obstruction occurred. CONCLUSIONS: LRN for WT and renal cancer may be considered as an option in selected children. Preoperative chemotherapy is to decrease tumor size and to facilitate the dissection, avoiding tumor rupture. For trained laparoscopic surgeons, the eventual indication of LRN is the tumor not crossing the midline. A long follow-up and more cases are necessary to evaluate the results of the laparoscopic approach.

Congenital mesoblastic nephroma: Clinical analysis of eight cases and a review of the literature.

Congenital mesoblastic nephroma (CMN) is a mesenchymal renal tumor. The aim of the present study was to review the clinical characteristics and outcome of CMN in infants. A retrospective file review was conducted of eight cases of CMN treated at the Children's Hospital of Fudan University between 2004 and 2012. Ultrasound and computerized tomography scans had been performed on all eight patients. Two cases presented with a solid tumor and exhibited pathological features consistent with those of classic CMN, five cases exhibited cystic, hemorrhagic and necrotic characteristics, with calcification and pathology consistent with the cellular variant of CMN and one case presented with a solid tumor, which exhibited pathological features consistent with ceullular CMN. Histology confirmed classic CMN in two patients and cellular CMN in six patients. For surgical intervention, four cases had radical nephrectomy, one case had a half nephrectomy and three cases had tumor enucleation performed. Two cases had received pre-operative chemotherapy, but exhibited no response, and three cases received post-operative chemotherapy. Two patients were lost to follow-up, but the remaining six patients survived to the end of follow-up without further complications. The mean follow-up time was 24.6 months. In conclusion, the differential diagnosis between CMN and Wilms' tumor is critical. Imaging characteristics are partially correlated with pathological characteristics. Surgery is the main treatment for CMN, but pre-operative chemotherapy is not particularly effective. The efficacy of post-operative chemotherapy requires further investigation, but the prognosis is positive.

Improved outcome of biliary atresia with postoperative high-dose steroid.

Objective. The dosage, duration, and the benefits of high-dose steroid treatment and outcome in biliary atresia (BA) remain controversial. In this study, we evaluated the impact of high-dose steroid therapy on the outcome of BA after the Kasai procedure. Methods. Intravenous prednisolone administration was started 1 week after surgery, followed by 8 to 12 weeks of oral prednisolone. Total bilirubin (TB) levels (3, 6, and 12 months after surgery), early onset of cholangitis, and two-year native liver survival were evaluated. Results. 53.4%, 56.9%, and 58.1% of the patients in the high-dose steroid group were jaundice-free 3, 6, and 12 months after surgery, respectively; these values were significantly higher than the 38.7%, 39.4%, and 43.3% of the low-dose steroid group. One year after surgery, the incidence of cholangitis in the high-dose group (32.0%) was lower than that in the low-dose group (48.0%). Infants with native liver in the high-dose group had a better two-year survival compared to those in the low-dose steroid group (53.7% versus 42.6%). Conclusions. The high-dose steroid protocol can reduce the incidence of cholangitis, increase the jaundice-free rate, and improve two-year survival with native liver after the Kasai operation.

[Recent incidences and trends of childhood malignant solid tumors in Shanghai, 2002-2010].

OBJECTIVE: To examine the recent incidences and trends of childhood malignant solid tumors in Shanghai. METHOD: Data from the population-based Shanghai Cancer Registry and related retrospective survey were used to analyze the patterns of incidence and trends of malignant solid tumors diagnosed between 2002 and 2010 in children aged 0-14 years. The distributions of incidences were described according to gender, age and cancer types which were classified according to International Classification of Childhood Cancer (ICCC). Annual age-standardized rates (ASRs) were adjusted by the world standard population. Approximate confidence intervals for standardized rate ratios (SRR) based Poisson distribution test-based methods were used to assess changes in incidence over the period 2002 - 2006 and 2007 - 2010. RESULT: (1)A total of 868 cases of childhood malignant solid tumors were diagnosed in Shanghai during 2002 - 2010, accounting for 65.8% of all childhood cancers. The ASR of 2002 - 2010 was 80.2 per million for all solid tumors. (2) The ASR was higher in boys (86.3 per million) than in girls (73.8 per million) with SRR 1.2 (95%CI 1.0 - 1.3). Incidence rate was the highest in the first five years of life with 93.4 per million. The age-specific incidence rates in 5 - 9 and 10 - 14 age groups were 65.2 and 79.3 per million, respectively. (3) CNS tumors, lymphomas, germ cell tumors, neuroblastoma, and soft tissue sarcomas were the top 5 most common solid tumors in children, with the incidence rate of 23.8, 11.0, 7.8, 7.7 and 6.8 per million, respectively. The patterns of subgroups varied in different age groups. Blastomas, such as neuroblastoma, retinoblastoma, were more common in the children aged 0 - 4 years, whereas epithelial carcinomas and bone tumors developed more frequently in elder children aged 10 - 14 years. (4) Compared with the ASR in 2002 - 2006, the ASR for both genders in 2007 - 2010 had no substantial changes (78.7 per million in 2002 - 2006 and 82.9 per million in 2007 - 2010). However, among boys, the incidence rate in 2007 - 2010 was significantly higher than that in 2002 - 2006 with SRR 1.2 (95%CI: 1.0 - 1.4). For specific subgroups of cancer, there were no substantial changes. Some cautions should be taken when interpreting results involving a small number of cases per year and those with wide 95% confidence intervals. CONCLUSION: The incidence rate of pediatric malignant solid tumors among males was higher than females during 2002 - 2010, and it differed among different age groups with the highest in the first five years of life. CNS tumor was the most common type of solid tumors in children. This was a unique characteristics comparing with adult reflected in disease spectrum and age of onset. The patterns of incidence and its trends for childhood malignant solid tumors in Shanghai could provide a basis for etiologic research and preventive interventions. The findings also suggest an urgent need for longer population-based surveillance to verify the pattern and changing trends.

[Effect of ambroxol on lung development of rat models with nitrofen-induced congenital diaphragmatic hernia].

OBJECTIVE: To investigate the effects of ambroxol on rat models with nitrofen-induced congenital diaphragmatic hernia (CDH) and its potential mechanism. METHODS: Nine pregnant female SD rats were randomly divided into 3 groups at Day 9.5: 2 ml olive oil intragastrically in control group (2 rats) and 200 mg nitrofen in nitrofen (2 rats) and ambroxol groups (5 rats). Antenatal ambroxol was given intraperitoneally to ambroxol group at Days 18.5, 19.5 and 20.5 of gestation while control and nitrofen groups only received intraperitoneal normal saline. At Day 21.5 the fetuses were delivered by cesarean section. Incidence of hernia, lung weight/body weight (LW/BW), mean terminal branch density (MTBD), percentage of lung alveolar area (PLAA), percentage of wall thickness (MT%) and the expression of TGF-ß1 were observed. RESULTS: There were 19 CDH fetuses in nitrofen group (68.4%). The incidence of hernia in ambroxol group was 65.1% (28/43). There was no significant difference (P>0.05) between two groups. LW/BW and PLAA decreased while MTBD and MT% increased significantly in the nitrofen group versus the control group [(45±6) mg/g vs (60±7) mg/g, (50.1±4.0)% vs (58.4±3.0)%, (14.0±1.8) vs (8.5±1.1), (45±6)% vs (29±6)%, all P=0.001]. After ambroxol intervention, the ambroxol group showed a higher PLAA but a lower MTBD and MT% [(54.0±2.0)%, (12.2±2.1), (39±4)%] than those in the nitrofen group (P=0.001, 0.006, 0.002). The expression of TGF-ß1 in pulmonary tissues of the nitrofen group was significantly higher than that in the control group (13,594±3113 vs 9447±1355, P=0.001). It decreased after ambroxol intervention (10 015±818, P=0.01). CONCLUSION: Though with no effect upon the occurrence of CDH in rats, the administration of ambroxol may improve the pulmonary maturity. The down-regulated expression of TGF-ß1 and the oxidative stability are possible mechanisms.

[Diagnosis and treatment of ovotesticular disorders of sex development in children].

OBJECTIVE: To investigate the diagnosis and treatment of ovotesticular disorders of sex development (DSD) in children. METHODS: We reviewed the clinical data of 9 cases of ovotesticular DSD admitted in our department from 1988 to 2007. RESULTS: The patients ranged in age from 9 months to 9 years, 7 raised as males and 2 as females. As for the karyotype, 4 cases were 46,XX, 2 were 46,XX/46,XY, 1 was 46,XY, and the other 2 had no karyotype data. All of them presented with obscure external genitalia: perineal or penoscrotal hypospadias with or without cryptorchidism in males and hypertrophy of the clitoris in females. They were diagnosed with ovotesticular DSD by gonad biopsy and underwent genitoplasty. CONCLUSION: The gender assignment of the ovotesticular DSD patient was chiefly based on the development of external genitalia, dominant gonad, karyotype and the parent's will. Laparoscopic technology is recommended in gonad biopsy and orchiopexy during the treatment of ovotesticular DSD.

The impact of iatrogenic gastroschisis on pulmonary maturation in the fetal rabbit models of congenital diaphragmatic hernia.

PURPOSE: The aim of this study was to analyze the effect of iatrogenic gastroschisis on pulmonary hypoplasia in fetal rabbits with congenital diaphragmatic hernia (CDH). MATERIALS AND METHODS: A total of 30 pregnant rabbits received fetal surgery on gestational day 23. A left diaphragmatic hernia was created in one end fetus (DH group) of each rabbit, and the other end fetus of the same rabbit received sham thoracotomy as control (CR group). Another 19 pregnant rabbits underwent partial resection of the diaphragm in both end fetuses on gestational day 23, and then artificial gastroschisis was performed on one end fetus (GS group) on gestational day 26, while the other end remained as control (CGS group). The fetuses were harvested on gestational day 30. The histological and morphometric evaluation of lungs and livers of the end fetuses in each group was conducted. RESULTS: In the DH group, the lungs were hypoplastic with a decrease in the total lung weight to body weight ratio, and remarkable thickening in alveolar septa. The lung vessels showed significantly thicker arterial walls when compared with those from control fetuses. The pathological finding in the CGS group was similar to that of the DH group. The thickness of the alveolar septa and of the pulmonary arterial walls showed no significant difference among the GS group, DH group and the CGS group. The ratio of liver weight to body weight increased notably in the GS group, DH group and CGS group compared with that in the CR group. CONCLUSIONS: In the fetal rabbit models of CDH, pulmonary hypoplasia is the most significant pathological feature. Iatrogenic gastroschisis does not improve pulmonary maturation due to the active growth of the liver that herniates into the thoracic cavity.

Relationship between prognosis of biliary atresia and infection of cytomegalovirus.

BACKGROUND: The etiology of biliary atresia is still unknown. It is generally accepted that virus infection may be one of the important causes that lead to biliary atresia. This study aimed to illustrate the relationship between infection of cytomegalovirus and prognosis of biliary atresia. METHODS: From January 2002 to March 2004, 27 patients who had undergone Kasai's procedure because of biliary atresia were investigated for cytomegalovirus -IgG, IgM and pp65, and their mothers were also examined for confirmation of cytomegalovirus infection. The patients were divided into three groups: infection free group, cytomegalovirus positive group and cytomegalovirus infection group. The rate of jaundice disappearance and the incidence of reflux cholangitis were analyzed statistically. The histopathological changes of the liver were also analyzed. RESULTS: The positive expression of cytomegalovirus -IgM and cytomegalovirus-pp65 in the patients was higher than that in their mothers (48% versus 14.81% and 37% versus 3.78%, respectively). Compared with the other two groups (80% in the infection free group, and 82% in the cytomegalovirus positive group), the rate of jaundice disappearance after operation in the cytomegalovirus infection group (36%) was significantly lower (P<0.05), and the incidence of reflux cholangitis was higher (P<0.05). Histopathological examination also showed that the degree of liver fibrosis and inflammation was more serious (P<0.05). CONCLUSIONS: There is a strong correlation between cytomegalovirus infection and a lower rate of jaundice disappearance, also a higher post-operational reflux cholangitis. Liver fibrosis seems to be more severe in biliary atresia patients with cytomegalovirus infection.

[Effects of zearalenone on the proliferation of SK-N-SH human neuroblastoma cells].

OBJECTIVE: To investigate effects of zearalenone (ZEA) on the proliferation of SK-N-SH human neuroblastoma cells in vitro and its possible mechanism. METHODS: SK-N-SH cells were cultured in estrogen-free improved minimum essential medium and divided into 5 groups based on different treatments: group 1, without treatment; group 2, treated with 17beta-estradiol (E(2)); group 3, treated with ZEA; group 4, treated with both E(2) and ICI 182780; group 5, treated with both ZEA and ICI 182780. Absorbance value (AV) was determined at the time point of 0, 24, 48 and 72 hours, and DNA proliferation index (PI) at 72 hours. Flow cytometer, terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) were employed to monitor cell apoptosis. RESULTS: At 24, 48 and 72 hours, the AV of group 3 were 1.39, 1.32, and 1.22 times to those of group 1, respectively. PI in group 3 was 1.43 times of that in group 1 at 72 hours. The results of group 2 were similar to those in group 3. At the same time, the growth of cells was inhibited by ICI 182780 despite the presence of E(2) and ZEA. Apoptosis cells were abundant in group 1 and ICI 182780 groups, but little in E(2) and ZEA groups. CONCLUSION: ZEA might promote the proliferation of SK-N-SH cells to a level similar to that of E(2), which might probably be brought about via estrogen receptor pathways and depressing apoptosis.