Whole-genome sequencing and RNA sequencing analysis reveals novel risk genes and differential expression patterns in hepatoblastoma.

Hepatoblastoma (HB) is an uncommon malignant liver cancer primarily affecting infants and children, characterized by the presence of tissue that resembling fetal hepatocytes, mature liver cells or bile duct cells. The primary symptom in affected children is abdominal lumps. HB constitutes approximately 28% of all liver tumors and two-thirds of liver malignancies in the pediatric and adolescent population. Despite its high prevalence, the underlying mechanism of HB pathogenesis remain largely unknown. To reveal the genetic alternations associated with HB, we conducted a comprehensive genomic study using whole-genome sequencing (WGS) and RNA sequencing (RNA-seq) techniques on five HB patients. We aimed to use WGS to identify somatic variant loci associated with HB, including single nucleotide polymorphisms (SNPs), insertions and deletions (Indels), and copy number variations (CNVs). Notably, we found deleterious mutation in CTNNB1, AXIN2 and PARP1, previously implicated in HB. In addition, we discovered multiple novel genes potentially associated with HB, including BRCA2 and GPC3 which require further functional validation to reveal their contributions to HB development. Furthermore, the American College of Medical Genetics and Genomics (ACMG) analysis identified the ABCC2 gene was the pathogenic gene as a potential risk gene linked with HB. To study the gene expression patterns in HB, we performed RNA-seq analysis and qPCR validation to reveal differential expression of four candidate genes (IGF1R, METTL1, AXIN2 and TP53) in tumors compared to nonneoplastic liver tissue in HB patients (P-Val < 0.01). These findings shed lights on the molecular mechanisms underlying HB development and facilitate to advance future personalized diagnosis and therapeutic interventions of HB.

In vitro and in vivo killing effects of methionine enkephalin on osteosarcoma.

OBJECTIVE: This study aimed to investigate the underlying regulatory effects of methionine enkephalin (MENK) on osteosarcoma. METHODS: The Cell Counting Kit-8 assay, clone formation, wound healing, transwell assay, and flow cytometry were performed to measure the effects of MENK on the proliferation, migration, invasion, and apoptosis of MG-63 and Saos-2 cells. Opiate growth factor receptor expression (OGFr) in cells was stably knocked down using siRNA. A tumor model was established by inoculating MG-63 cells into mice. Flow cytometry was performed to identify alterations in mice bone marrow, spleen, and tumor tissue immune cells. The phenotype of tumor-associated macrophages was determined using immunohistochemistry. After OGFr knockdown or/and treatment with MENK, Bax, Bcl-2, caspase 3, caspase 9, and PARP expression levels were characterized using qRT-PCR, western blot, and WES, respectively. RESULTS: MENK could significantly inhibit the proliferation, invasion, and migration of MG-63 and Saos-2, arrest the cell cycle in the G0/G1 phase, upregulate Bax, caspase 3, caspase 9, and PARP expression, and downregulate Bcl-2 expression. Tumor size and weight were lower in the MENK group than those in the control group. MENK-treated mice exhibited a reduced ratio of CD11b + Gr-1 + myeloid-derived suppressor cells. MENK increased the ratio of M1-type macrophages and decreased the proportion of M2-type macrophages in tumor tissue. Furthermore, the level of TNF-α significantly increased while that of IL-10 decreased in MENK-treated mice. The effect of MENK could be partly reversed by OGFr knockdown. CONCLUSION: MENK reduces the abundance of myeloid-derived suppressor cells, induces M1 polarization of macrophages, and exhibits an inhibitory effect on osteosarcoma.

Accumulation and migration of microplastics and its influencing factors in coastal saline-alkali soils amended with sewage sludge.

Coastal saline-alkali soil can be transformed to agricultural soil with sewage sludge amendment. However, sewage sludge contains a large number of microplastics (MPs), and the fate of MPs in sludge-treated saline-alkali soil needs to be studied. Therefore, we investigated the accumulation and migration of MPs, and their influencing factors in saline-alkali soil after one-time sewage sludge application (0, 25, 50, 100 and 200 t ha-1 SSA). The results indicated that sewage sludge input contributed to MP accumulation in soil, and the MP abundance in 20-40 cm soil was significantly lower than that in 0-20 cm soil. Fragments and fibers were the most abundant MPs in soil, and the proportions of fragments and 50-200 µm MPs in 20-40 cm soil were lower than those in 0-20 cm soil, while the < 50 µm MP proportion was higher than that in 0-20 cm soil. Correlation analysis showed that MP accumulation rate (0-40 cm) and migration rate (20-40 cm) were negatively correlated with soil organic matter (SOM) content and SSA, but positively correlated with soil pH. Stepwise regression analysis further showed that SOM and SSA were the main factors affecting MP accumulation rate, which explained 47.7% and 46% of its variation, respectively, while pH was the crucial factor affecting the migration rate of MPs, followed by EC and SSA. In conclusion, SSA caused MP accumulation in saline-alkali soil, and SSA primarily affected the MP abundance, while soil OM, pH and EC directly affected MP migration in soil.

Protocol to dissociate and isolate wide-diversity single cells by density gradient centrifugation from human hepatoblastoma tissue.

Single-cell transcriptome sequencing can characterize various cell types in human liver tissue and facilitate understanding of hepatoblastoma heterogeneity. Here, we present a protocol for isolating hepatocytes and immune cells from human hepatoblastoma samples with high viability. We describe steps for tissue processing, enzymatic digestion, Percoll density gradient separation, cell lysis, cell suspension quality control, and scRNA library construction. We then detail sequencing and data analysis. This protocol is applicable to preparing single-cell suspensions from other human liver tissue samples.

Zygomaticotemporal suture maturation evaluation in Chinese population using cone-beam computed tomography images.

Objective: This study aimed to evaluate the zygomaticotemporal suture (ZTS) maturation, analyze the age distribution patterns of ZTS maturation stages, and investigate the relationship between ZTS and cervical vertebral maturation (CVM). Methods: A total of 261 patients who underwent cone-beam computed tomography (112 males, mean age, 13.1 ± 3.3 years; 149 females, mean age, 13.7 ± 3.1 years) were examined to evaluate the ZTS stages. The ZTS stages were defined based on a modified method from previous studies on zygomaticomaxillary sutures. Differences between groups and correlations between indicators were analyzed using the Spearman correlation test, intraclass coefficient of correlation (ICC), one-way analysis of variance and rank sum test. Statistical significance was set at p < 0.05. The diagnostic value of CVM stages in identifying ZTS maturation stages was evaluated using positive likelihood ratios (LRs). Results: A positive relationship was found between the ZTS and CVM stage (r = 0.747, ICC = 0.621, p < 0.01) and between the ZTS stage and chronological age (r = 0.727, ICC = 0.330, p < 0.01). Positive LRs > 10 were found for several cervical stages (CSs), including CS1 and CS2 for the diagnosis of stage B, CS1 to CS3 for the diagnosis of stages B and C, and CS6 for the diagnosis of stages D and E. Conclusions: The ZTS maturation stage may be more relevant to the CVM stage than to the chronological age. The CVM stages can be good indicators for clinical decisions regarding maxillary protraction, except for CS4 and CS5.

ACLY-ß-catenin axis modulates hepatoblastoma cell proliferation.

HB (hepatoblastoma) is most common in children with liver cancer and few options for treating HB. Thus, it is of great significance to investigate the regulatory mechanism of HB and/or identify new therapeutic targets for clinical treatment of HB. Here, we showed that ACLY (ATP citrate lyase), an important lipometabolic enzyme for de novo biosynthesis of fatty acids and steroids, has a higher expression in HB tissues than noncancerous tissues, and is required for HB cell proliferation. Moreover, knocking down ACLY in HB cells caused severe S-phase arrest and apoptosis. Mechanistically, ACLY knockdown significantly silenced the Wnt signaling pathway and reduced ß-catenin expression in HB cells. Conversely, the apoptotic alleviation of HB cells by overexpressing ACLY was blocked by silencing ß-catenin, suggesting the modulation of HB cells by ACLY-ß-catenin axis. Our results uncovered the role of ACLY in HB cells and presented a theoretical approach for HB targeted therapy in the future.

O-GlcNAcylated LARP1 positively regulated by circCLNS1A facilitates hepatoblastoma progression through DKK4/ß-catenin signalling.

BACKGROUND: Accumulating studies have shown that La-related protein 1 (LARP1) is involved in the occurrence and development of various tumours. However, the expression pattern and biological role of LARP1 in hepatoblastoma (HB) remain unclear so far. METHODS: LARP1 expression level in HB and adjacent normal liver tissues was analysed by qRT-PCR, Western blotting and immunohistochemistry assays. The prognostic significance of LARP1 was evaluated by Kaplan-Meier method and multivariate Cox regression analysis. In vitro and in vivo functional assays were implemented to clarify the biological effects of LARP1 on HB cells. Mechanistically, the regulatory roles of O-GlcNAcylation and circCLNS1A in LARP1 expression were investigated by co-immunoprecipitation (co-IP), immunofluorescence, RNA immunoprecipitation (RIP), RNA pull-down and protein stability assays. Moreover, RNA-sequencing, co-IP, RIP, mRNA stability and poly(A)-tail length assays were performed to investigate the association between LARP1 and DKK4. The expression and diagnostic significance of plasma DKK4 protein in multi-centre cohorts were evaluated by ELISA and ROC curves. RESULTS: LARP1 mRNA and protein levels were remarkably elevated in HB tissues and associated with worse prognosis of HB patients. LARP1 knockdown abolished cell proliferation, triggered cell apoptosis in vitro as well as prohibited tumour growth in vivo, whereas LARP1 overexpression incited HB progression. Mechanistically, O-GlcNAcylation of LARP1 Ser672 by O-GlcNAc transferase strengthened its binding to circCLNS1A and then protected LARP1 from TRIM-25-mediated ubiquitination and proteolysis. LARP1 upregulation subsequently led to DKK4 mRNA stabilisation by competitively interacting with PABPC1 to prevent DKK4 mRNA from B-cell translocation gene 2-dependent deadenylation and degradation, thus facilitating ß-catenin protein expression and nuclear import. CONCLUSION: This study indicates that upregulated protein level of O-GlcNAcylated LARP1 mediated by circCLNS1A promotes the tumorigenesis and progression of HB through LARP1/DKK4/ß-catenin axis. Hence, LARP1 and DKK4 are promising therapeutical target and diagnostic/prognostic plasma biomarker for HB.

Conditional remediation performance of wheat straw biochar on three typical Cd-contaminated soils.

Undoubtedly biochar has excellent remediation performance for Cd-contaminated soil. Nevertheless, the remediation performance may be not invariable considering highly variable soil conditions including soil properties and environmental conditions. This work investigated the fate of Cd in three typical Cd-contaminated soils (acidic, neutral and saline-alkali soils) treated with wheat straw biochar and its driving mechanisms under specific soil conditions through aging and remediation experiment, Cd availability experiment and leaching column experiment. The results indicated that biochar addition facilitated Cd immobilization and reduced the uptake of Cd by green vegetables in acidic, neutral and saline-alkali soils under wetting-drying conditions. In contrast to neutral and saline-alkali soils, the release of exchangeable aluminum from biochar-treated acidic soil under flooding-drying cycles lowered the pH of leachate, thus promoting the leaching of Cd from leaching column, especially at 7 and 14 days, when the leaching of Cd increased by 25.3 and 32.6 times, respectively. This result was further supported by the increase in the exchangeable fraction and total leaching amounts of Cd in the topsoil layer (0-20 cm) of biochar-treated acidic soil of leaching column. Additionally, the leaching of Cd was positively correlated with DOC contents of leachate in biochar-treated neutral and saline-alkali soils. In summary, the remediation performance of biochar for Cd-contaminated soils is conditional, and its remediation effect is better in neutral and saline-alkali soils. Notably, the inherent conditions of soil must be fully considered when applying biochar for Cd remediation, especially in acidic Cd-contaminated paddy soils in South China.

Zygomaticomaxillary suture maturation evaluation in patients with and without cleft lip and palate.

INTRODUCTION: The zygomaticomaxillary suture (ZMS) maturation evaluation is a reliable method for predicting the optimal timing of maxillary protraction. The objective of this study was to compare age distribution patterns of ZMS maturation stages between cleft lip and palate (CLP) patients and non-cleft lip and palate (non-CLP) patients to aid our comprehension in choosing the optimal timing of maxillary protraction. METHODS: Samples of 216 non-CLP and 220 CLP Asian patients without orthodontic and orthognathic treatment aged 5-25 years were scanned to evaluate the ZMS maturation stage by 2 evaluators blindly. Evaluators' agreements and bilateral ZMS maturation consistency were assessed by weighted kappa tests. Age distribution patterns of each ZMS maturation stage were described. Gender effect and age distribution differences between groups were analyzed using an independent t-test. RESULTS: Evaluators' agreements and bilateral ZMS maturation consistency were satisfying (weighted kappa coefficient >0.90). At stages A and B, patients with CLP were 1.3 and 0.4 years older than patients in the non-CLP group (P <0.001 and P = 0.01). In contrast, at stage C, patients with CLP were approximately 1.2 years younger (P = 0.004). Gender barely played a role in the divergence of ZMS maturation (P >0.05). No statistically significant difference was observed between ZMS maturation of patients with unilateral or bilateral cleft lip and palate (UBCLP) and patients with unilateral or bilateral cleft lip (UBCL) (P >0.05). CONCLUSIONS: The ZMS development of patients with CLP was premature at stage C, whereas delayed at stages A and B.

Structural and Predicted Functional Diversities of Bacterial Microbiome in Response to Sewage Sludge Amendment in Coastal Mudflat Soil.

The study investigated the influence of sewage sludge application at rates of 0 (CK), 30 (ST), 75 (MT), and 150 (HT) t ha-1 to mudflats on bacterial community diversity and predicted functions using amplicon-based sequencing. Soils under sewage sludge treatments, especially the HT treatment, exhibited lower pH, salinity and higher nutrient contents (C, N, and P). Moreover, restructured bacterial communities with significantly higher diversities and distinct core and unique microbiomes were observed in all sewage sludge-amended soils as compared to the control. Specifically, core bacterial families, such as Hyphomicrobiaceae, Cytophagaceae, Pirellulaceae Microbacteriaceae, and Phyllobacteriaceae, were significantly enriched in sewage sludge-amended soils. In addition, sewage sludge amendment significantly improved predicted functional diversities of core microbiomes, with significantly higher accumulative relative abundances of functions related to carbon and nitrogen cycling processes compared to the unamended treatment. Correlation analyses showed that modified soil physicochemical properties were conducive for the improvement of diversities of bacterial communities and predicted functionalities. These outcomes demonstrated that sewage sludge amendment not only alleviated saline-sodic and nutrient deficiency conditions, but also restructured bacterial communities with higher diversities and versatile functions, which may be particularly important for the fertility formation and development of mudflat soils.

An analysis of the diagnosis, clinical characteristics, treatment, and survival outcomes of 36 extracranial malignant rhabdoid tumor patients.

BACKGROUND: Extracranial malignant Rhabdoid tumors (eMRTs) are rare but aggressive lesions in young children. This work aimed to review and analyze the diagnosis, clinical characteristics, treatment, and survival of eMRTs so as to summarize experience for future therapy. METHODS: A total of 36 eMRT cases were treated between January 2008 and August 2019 according to Shanghai Children's Medical Center (SCMC) multimodal protocol of mixed surgery, radiation and chemotherapy involving vincristine, carboplatin, doxorubicin, etoposide, cyclophosphamide and ifosfamide. We collected information including: age at diagnosis, tumor location, disease stage, therapy, outcomes, etc. Overall survival (OS) and event free survival (EFS) were calculated and risk factors for survival were analyzed. RESULTS: The patients had a median age of 1.80 years at diagnosis (range, 1.4 m-13.42 years), and were followed up for 9.17 months in median (range, 4 d-11.14 y). A total of 16 patients achieved complete remission (CR), and 7 survived without reoccurrence till December 2019. The 3-year EFS was 17.4% (95% CI: 11.0-23.8%) with a 3-year OS of 23.4% (95% CI: 15.8-31.0%). Recurrence was found only in children younger than the median age (1.80 y). Localized staging (Log Rank P=0.039 for OS and P=0.021) and older age (Log Rank P=0.016 for OS and P=0.002 for EFS) were associated with improved outcome. Younger age (Cox regression, OS, OR =2.610, 95% CI: 1.147-5.937, P=0.022; EFS, OR =3.401, 95% CI: 1.495-7.752, P=0.004) were independent risk factors for death and recurrence. CONCLUSIONS: Those eMRTs treated according to SCMC protocol turned out to have poor outcomes. Higher staging at diagnosis and reoccurrence in younger patients remain major threats to the prognosis.

Sirolimus for the treatment of kaposiform hemangioendothelioma: In a trough level-dependent way.

With the accumulation of clinical practice, sirolimus is now widely viewed as an effective agent in kaposiform hemangioendothelioma (KHE) treatment using a dose based on experience. Therefore, this retrospective research aimed to provide evidence-based suggestions on the most appropriate dose and trough level of sirolimus. All unresectable KHE cases diagnosed at our center from January 2016 to December 2019 were included. Sirolimus monotherapy was initiated when there was no sign of Kasabach-Merritt phenomenon (KMP) at a dose of 0.8 mg/m2 twice a day in order to keep the trough level at 5-20 ng/mL. Patients' clinical information, tumor volume change, trough level fluctuation, and complication occurrence were all recorded. Efficacy represented by tumor shrinkage speed and safety manifested by complication grades were compared between different trough level groups (5-10 vs. 10-15 vs. >15 ng/mL). Twenty-one patients (10 girls and 11 boys) were enrolled. There were eight patients in the 5-10 ng/mL group, seven in the 10-15 ng/mL group, and six in the more than 15 ng/mL group. Trough level over 10 ng/mL manifested better efficacy in tumor shrinkage (t-test, p = 0.011) while a level over 15 ng/mL had no further benefit in efficacy (t-test, p = 0.65). In addition, tumors at a central location reacted better to sirolimus (t-test, p = 0.022). No significant differences were observed in complication occurrence among different concentrations, although boys seemed to be at higher risk of more severe complications (>grade II, χ2 -test, p = 0.009, odds ratio = 4.52, range = 1.20-17.24). It proved to be most efficacious in the management of KHE at a trough level between 10 and 15 ng/mL. Such concentration was safe and well tolerated.

The Fibroblast Growth Factor 9 (Fgf9) Participates in Palatogenesis by Promoting Palatal Growth and Elevation.

Cleft palate, a common global congenital malformation, occurs due to disturbances in palatal growth, elevation, contact, and fusion during palatogenesis. The Fibroblast growth factor 9 (FGF9) mutation has been discovered in humans with cleft lip and palate. Fgf9 is expressed in both the epithelium and mesenchyme, with temporospatial diversity during palatogenesis. However, the specific role of Fgf9 in palatogenesis has not been extensively discussed. Herein, we used Ddx4-Cre mice to generate an Fgf9-/- mouse model (with an Fgf9 exon 2 deletion) that exhibited a craniofacial syndrome involving a cleft palate and deficient mandibular size with 100% penetrance. A smaller palatal shelf size, delayed palatal elevation, and contact failure were investigated to be the intrinsic causes for cleft palate. Hyaluronic acid accumulation in the extracellular matrix (ECM) sharply decreased, while the cell density correspondingly increased in Fgf9-/- mice. Additionally, significant decreases in cell proliferation were discovered in not only the palatal epithelium and mesenchyme but also among cells in Meckel's cartilage and around the mandibular bone in Fgf9-/- mice. Serial sections of embryonic heads dissected at embryonic day 14.5 (E14.5) were subjected to craniofacial morphometric measurement. This highlighted the reduced oral volume owing to abnormal tongue size and descent, and insufficient mandibular size, which disturbed palatal elevation in Fgf9-/- mice. These results indicate that Fgf9 facilitates palatal growth and timely elevation by regulating cell proliferation and hyaluronic acid accumulation. Moreover, Fgf9 ensures that the palatal elevation process has adequate space by influencing tongue descent, tongue morphology, and mandibular growth.

Inflammatory myofibroblastic tumor successfully treated with metformin: A case report and review of literature.

BACKGROUND: Inflammatory myofibroblastic tumor (IMT) is a distinct tumor with a low incidence rate, which can be diagnosed at any age with a predilection for children and adolescents. Although IMT is visible in any tissues and organs, it is more commonly found in the lungs. The clinical and radiological manifestations of IMT lack specificity, hence resulting in frequent misdiagnosis. Surgical resection is currently the main therapeutic approach for IMT. Only scarce cases of IMT treated with metformin have been reported. Here we report the case of an IMT patient with partial penile resection treated with metformin. CASE SUMMARY: A 1-year-old boy was born with a shorter penis, and his foreskin could not be completely turned over. When he was 6 month old, a well-circumscribed mass on the glans was found, while it did not attract the attention of his parents. The mass gradually increased in size over time before he was admitted to the hospital, where physical examination was performed. It was revealed that the glans hidden behind the foreskin had a mass with a diameter of about 4 cm surrounding the penis. The mass appeared to be hard with a smooth surface and poor mobility. The two testicles examined at the bottom of the scrotum were revealed to have a normal size. Magnetic resonance imaging showed a tumor with rich blood supply encircling the cavernosum with a size of 3.5 cm × 2.1 cm × 2.0 cm. A thick urinary line was found without urine dripping, urgency, and urodynia. Surgical treatment was performed. During the operation, it was observed that the mass had surrounded and invaded the cavernosum without obvious boundaries, and that the tumor occupied about one-half of the penis cross-section as well as infiltrated more than one-half of the glans. In addition, the tumor had caused urethral invasion and anterior urethrostenosis. With the intention of keeping the glans and cavernosum, the tumor at the anterior urethra was partially removed, leaving about 30% of the tumor mass. Pathology analysis demonstrated that the tumor was rich in spindle cells with infiltration of inflammatory cells. Immuno-histochemistry analysis indicated that the cells were positive for CD4, CD99, Ki67, BCL2, and CD68, and negative for ALK, MyoG, S100, SOX10, PR, and EMA. Hence, the tumor was diagnosed as IMT. Metformin was prescribed for the patient after the operation, following which an oral dose of 7 mg/kg was given three times a day after meals. Three months later, it was observed that the remaining tumor had completely disappeared and that the urination process from the urethra opening had resumed normal. In addition, there were no side effects observed. There was also no tumor recurrence. The growth and development of the boy were unaffected as a result of the treatment. CONCLUSION: The tumor was observed to have completely disappeared after treatment with metformin. Our finding is of great significance to facilitate future clinical treatment with IMT.

Sewage sludge application enhances soil properties and rice growth in a salt-affected mudflat soil.

The most important measures for salt-affected mudflat soil reclamation are to reduce salinity and to increase soil organic carbon (OC) content and thus soil fertility. Salinity reduction is often accomplished through costly freshwater irrigation by special engineering measures. Whether fertility enhancement only through one-off application of a great amount of OC can improve soil properties and promote plant growth in salt-affected mudflat soil remains unclear. Therefore, the objective of our indoor pot experiment was to study the effects of OC amendment at 0, 0.5%, 1.0%, 1.5%, and 2.5%, calculated from carbon content, by one-off application of sewage sludge on soil properties, rice yield, and root growth in salt-affected mudflat soil under waterlogged conditions. The results showed that the application of sewage sludge promoted soil fertility by reducing soil pH and increasing content of OC, nitrogen and phosphorus in salt-affected mudflat soil, while soil electric conductivity (EC) increased with increasing sewage sludge (SS) application rates under waterlogged conditions. In this study, the rice growth was not inhibited by the highest EC of 4.43 dS m-1 even at high doses of SS application. The SS application increased yield of rice, promoted root growth, enhanced root activity and root flux activity, and increased the soluble sugar and amino acid content in the bleeding sap of rice plants at the tillering, jointing, and maturity stages. In conclusion, fertility enhancement through organic carbon amendment can "offset" the adverse effects of increased salinity and promote plant growth in salt-affected mudflat soil under waterlogged conditions.

Sustained effects of one-time sewage sludge addition on rice yield and heavy metals accumulation in salt-affected mudflat soil.

High-yielding and sustainable production of rice in salt-affected mudflat is restricted by high soil salinity. Although sewage sludge can be used for mudflat amendment especially soil salt reduction, the possibility of potential heavy metal contamination in sludge-amended mudflat especially under paddy cultivation remains unclear, which hinders the further utilization of sewage sludge. In this study, a field experiment was conducted in a newly reclaimed mudflat to assess the sustained effects of one-time sludge input with different addition rates (0, 30, 60, 120, and 180 t ha-1) on soil salinity, rice yield, and potential metal contamination under paddy cultivation. The results indicated that sewage sludge addition (SSA) significantly decreased soil salinity and increased soil fertility. The increasing SSA rates and amending years led to the gradual increase of rice yield in salt-affected mudflat. The maximum increases in rice yield were 125.1%, 124.7%, and 127.9% in 2016, 2017, and 2018, and the average annual increase in rice yield in sludge-treated mudflat was 1.7%. Sludge addition increased metals accumulation in mudflat soil and metals uptake by rice tissues except Cr, Cu, and Pb in rice grain. The maximum increments in metal concentrations in soil and rice plant all occurred at 180 t ha-1 sludge addition rate. However, the metal concentrations in rice grain were below the safety limits even in the treatment with the highest sludge addition rate. Metal concentrations in sludge-treated soil and rice plant showed downward trend during the 3-year trial, and the decreases in total amount of soil metals were mainly concentrated in the first amending year, accounting for more than 50%. In summary, one-time sludge input achieved sustained mudflat amendment and efficient rice production. In addition, controlling the total amount of sludge input realized safe utilization of sewage sludge in salt-affected mudflat under paddy cultivation.

Melanotic neuroectodermal tumor of infancy successfully treated with metformin: A case report.

RATIONALE: Melanotic neuroectodermal tumor of infancy (MNTI) is a rare tumor originated from neural crest cells with the potential for recurrence and metastasis. The peak age for the disease is during the first year after birth. The current therapy is primarily surgery. The patient reported here is the first case of MNTI treated with metformin. PATIENT CONCERNS: A case of a 4-month-old infant with a history of swelling in the mouth for 1 month. DIAGNOSIS: The tumor was diagnosed using radiology, pathology, and immunohistochemistry, and it was performed with complete surgical resection. Unfortunately, the tumor recurred 3 months after surgery. INTERVENTIONS: We prescribed metformin for the infant. OUTCOMES: Currently, after 9 months of treatment, the tumor is well controlled without apparent side effects. LESSONS: The case presented suggested that metformin may be an underlying therapy for MNTI.

Gorham-Stout disease successfully treated with sirolimus (rapamycin): a case report and review of the literature.

BACKGROUND: Gorham-Stout disease (GSD) is a rare disease characterized by bone lesions and osteolysis. Therapy usually involves surgical resection. Sirolimus (Rapamycin) is used in some patients with GSD but the efficacy and safety of Sirolimus remains unclear. We propose that Sirolimus may be a novel therapeutic for GSD and present a case and review of literature that supports this. CASE PRESENTATION: We presented a 1-year-old boy with GSD involving osteolysis of the right humerus with fracture of the left femur complicated by an effusion in the right pleural cavity. X-rays showed osteolysis in the right clavicle. A large pleural effusion was observed on the right-side, and the left lung was significantly compressed. X-rays also showed a fracture of the left femur. A femoral biopsy was performed that showed necrotic tissue in the cortical bone and a large number of irregularly shaped capillaries that proliferated within the necrotic tissue. Dilated lymphatic vessels were seen adjacent to the cortex, with fibrous tissue hyperplasia. We prescribed sirolimus, which is an oral mTOR inhibitor, for two consecutive years. The boy recovered well without other progressive bone lesions and participates in normal daily activities. His growth and development are the same as that of his peers. DISCUSSION AND CONCLUSION: Gorham-Stout disease is a rare and enigmatic disease characterized by the presentation of an intraosseous lymphatic anomaly (LM), which results in progressive bone resorption. Based on this case report and a literature review, we conclude that sirolimus may be an effective alternative medication for GSD.

Propranolol for infantile hepatic hemangioendothelioma: Clinical evaluation of drug efficacy and safety using a single-center patient cohort.

INTRODUCTION AND OBJECTIVES: Infantile hepatic hemangioendothelioma (IHHE) is a benign liver tumor, associated with hypothyroidism and vascular malformations along the skin, brain, digestive tract and other organs. Here, we determined a single-center patient cohort by evaluating the effectiveness and safety of propranolol and sirolimus for the treatment of IHHE. PATIENTS AND METHODS: We performed a monocentric and observational study, based on clinical data obtained from 20 cases of IHHE treated with oral propranolol and sirolimus at the Shanghai Children's Medical Center (SCMC), between December 2017 and April 2019. All cases were confirmed by abdominal enhanced CT examination (18/20, 90%) and sustained decrease of alpha fetoprotein (AFP) (2/20, 10%). Propranolol treatment was standardized as once a day at 1.0mg/kg for patients younger than 2 months, and twice a day at 1.0mg/kg (per dose) for patients older than 2 months. Sirolimus was used to treat refractory IHHE patients after 6 months of propranolol treatment, and initial dosing was at 0.8mg/m2 body surface per dose, administered every 12h. Upon treatment, abdominal ultrasound scanning was regularly performed to evaluate any therapeutic effects. All children were followed up for 6-22 months (mean value of 12.75 months). The clinical manifestations and therapeutic effects, including complications during drug management, were reviewed after periodic follow-up. RESULTS: The effective rate of propranolol for the treatment of children with IHHE was 85% (17/20). In most cases, the AFP levels gradually decreased into the normal range. A complete response (CR) was achieved in 3 cases, partial response (PR) for 14 cases, progressive disease (PD) for 2 cases and stable disease (SD) was only detected once. Lesions decreased in two PD patients after administration of oral sirolimus. No serious adverse reactions were observed. CONCLUSION: This study indicates that both propranolol and sirolimus were effective drugs for the treatment of children with IHHE at SCMC.