The methyltransferase METTL3 regulates endothelial cell proliferation and inflammation via m6A RNA methylation-mediated TRAF1 expression.

The imbalance of vascular endothelial cell homeostasis is the key mechanism for the progression of many vascular diseases. RNA modification, particularly N6-Methyladenosine (m6A), plays important function in numerous biological processes. Nevertheless, the regulatory function of m6A RNA methylation in endothelial dysfunction remains insufficiently characterized. In this study, we established that the m6A methyltransferase METTL3 is critical for regulating endothelial function. Functionally, depletion of METTL3 results in decreased endothelial cells proliferation, survival and inflammatory response. Conversely, overexpression of METTL3 elicited the opposite effects. Mechanistically, MeRIP-seq identified that METTL3 catalyzed m6A modification of TRAF1 mRNA and enhanced TRAF1 translation, thereby up-regulation of TRAF1 protein. Over-expression of TRAF1 successfully rescued the inhibition of proliferation and adhesion of endothelial cells due to METTL3 knockdown. Additionally, m6A methylation-mediated TRAF1 expression can be reversed by the demethylase ALKBH5. Knockdown of ALKBH5 upregulated the level of m6A and protein level of TRAF1, and also increased endothelial cells adhesion and inflammatory response. Collectively, our findings suggest that METTL3 regulates vascular endothelium homeostasis through TRAF1 m6A modification, suggesting that targeting the METTL3-m6A-TRAF1 axis may hold therapeutic potential for patients with vascular diseases.

Application of femoral nail, reconstruction locked plate and bone morphogenic protein - 7 in an osteopetrosis patient with a recurrent femoral shaft fracture: A case report.

INTRODUCTION AND IMPORTANCE: Femoral fractures are common in the patients with osteopetrosis and multiple treatment strategies have been described with varying results. However, there is a paucity of literature describing the treatment of recurrent fractures and subsequent deformity. CASE PRESENTATION: We present detailed revision strategies and long-term follow-up results of a patient with osteopetrosis who suffered unsuccessful operative treatment using the plate-screw system (recurrent femoral shaft fracture and implant failure). CLINICAL DISCUSSION: The success of revision surgery of osteopetrosis is based on good preoperative planning, appropriate selection of fixation methods, and a meticulous approach during surgery. The combined application of the expert adolescent lateral femoral nail, the reconstruction locked plate, and bone morphogenic protein (BMP)-7 in this patient achieved good clinical results. CONCLUSION: In the treatment of failed plated and recurrent osteopetrotic femoral shaft fractures, the combination of nails and plating presents an alternative, potentially more successful, revision strategy.

Effects of the nitrate and ammonium ratio on plant characteristics and Erythropalum scandens Bl. substrates.

Erythropalum scandens Bl. is a woody vegetable with high nitrogen demand that inhabits southern China. Ammonium and nitrate are the two main forms of inorganic nitrogen that plants directly absorb. A pot experiment was performed to determine the growth, physiological responses, and preferences of 12-month-old E. scandens seedlings for ammonium and nitrate. Aboveground and underground growth indexes, biomass, physiological and biochemical indexes (chlorophyll [Chl], soluble sugar, soluble protein and free proline contents), and substrate pH and nitrogen contents were determined under different nitrate and ammonium ratios (0 NO3-: 100 NH4+, 25 NO3-: 75 NH4+, 50 NO3-: 50 NH4+, 75 NO3-: 25 NH4+, and 100 NO3-: 0 NH4+), and the control (0 NO3-: 0 NH4+). The results showed that ammonium and nitrate improved the growth and physiological status of E. scandens seedlings in most of the treatments compared to the control. The aboveground growth status and biomass accumulation of E. scandens seedlings were significantly better under the 0 NO3-: 100 NH4+ treatment during fertilization compared with all other treatments. However, the growth status of the underground parts was not significantly different among treatments. Significant differences in osmoregulator content, except for soluble sugars, and Chl content were observed. Soluble sugars and soluble proteins were highest under the 0 NO3-: 100 NH4+ treatment at the end of fertilization (day 175). However, free proline accumulated during fertilization and the increase in NO3- indicated that excessive use of NO3- had a negative effect on the E. scandens seedlings. The order of accumulating nitrogen content was leaves > roots > stems. The highest N accumulation occurred in the aboveground parts under the 0 NO3-: 100 NH4+ treatment, whereas the highest N accumulation occurred in the underground parts under the 50 NO3-: 50 NH4+ treatment. Substrate pH increased at the end of fertilization (day 175) compared with the middle stage (day 75), while total nitrogen, ammonium, and nitrate were highly significantly different among the treatments. Total nitrogen and NH4+ content were the highest under the 0 NO3-: 100 NH4+ treatment, while NO3- content was the highest under the 100 NO3-: 0 NH4+ treatment. In conclusion, 12-month-old E. scandens seedlings grew best, and had better physiological conditions in NH4+ than NO3-. The 0 NO3-:100 NH4+ treatment (ammonium chloride 3.82 g/plant) resulted in the best growth and physiological conditions. Most of the growth and physiological indexes were inhibited with the increase in nitrate.

Resistant distal femoral nonunion treated with combined nail/plate construct and reamer-irrigator-aspirator technique.

OBJECTIVE: This study was performed to evaluate the effectiveness of intramedullary nailing and a lateral locking plate combined with the reamer-irrigator-aspirator (RIA) bone grafting technique for resistant distal femoral nonunion. METHODS: This retrospective observational study was performed from January 2018 to December 2021 and involved five patients who presented with resistant distal femoral nonunion despite undergoing several surgeries. They were treated with intramedullary nailing and a lateral locking plate combined with the RIA bone grafting technique. Postoperative follow-up was performed to observe the healing time, and functional outcomes were evaluated using the Lower Extremity Functional Scale (LEFS). RESULTS: After the patients had been monitored for a mean of 17.9 months, complete bone healing was observed in every patient (mean healing time of 4.8 months). Postoperative wound failure in an older patient was successfully treated with resuturing and nutritional assistance. At the last follow-up, the mean LEFS score was 71.2/80 and the mean knee flexion was 109 degrees. CONCLUSIONS: Our study demonstrates that combining intramedullary nailing and a lateral locking plate with the RIA bone grafting technique enhances biological properties, provides good structural support, and achieves good union and functional results in the management of resistant nonunion of the distal femur.

A rare presentation of type II Abernethy malformation and nephrotic syndrome: Case report and review.

Type II Abernethy malformation is an extremely reported congenital extrahepatic portosystemic shunt in complication with nephrotic syndrome. We present the case of an 8-year-old boy who presented with symptoms of type II Abernethy malformation and nephrotic syndrome. This diagnosis of this type II Abernethy malformation was based on physical examination, blood tests, urinalysis, nephrotic and hepatic function tests, routine clinical lipid measurements, abdominal ultrasonography, and computed tomographic angiography. A kidney biopsy revealed the pathological features of nephrotic syndrome. This is the second reported patient diagnosed with type II Abernethy malformation and nephrotic syndrome. Captopril treatment was effective in improving the symptoms of this case. A patient with type II Abernethy malformation related to immune complex-mediated glomerular injury was effectively improved with medication. Type II Abernethy malformation is a causative factor of immune complex-mediated glomerular injury in nephrotic syndrome. Captopril treatment significantly improved the symptoms in this case.

The Stability of Intact Parathyroid Hormone (PTH) in Different Types of Blood Collection Tubes.

BACKGROUND: Over past decades, the instability of parathyroid hormone (PTH) causes great interference for the clinical laboratory. Contradictory results were reported in many reports about storage conditions and suitable blood collection tubes to ensure PTH stability in the pretreatment phase. METHODS: This study recruited 30 participants including 10 healthy persons, 10 hemodialysis, and 10 hyperparathyroidism patients. Five types of blood collection tubes (EDTA-K3 tube, coagulant tube, heparin anticoagulant tube, gel separating tube, and plain tube) were included to determine whether they were suitable as blood-collecting vessels. The time points and conditions for testing samples included less than 2 hours, 4 hours, and 8 hours at room temperature, and, in parallel, 24 hours, 48 hours, and 72 hours in refrigeration. Two different judgement criteria were used to compare the stability of PTH in different blood vessels. RESULTS: Purely statistical analysis showed that 4 types of blood collection tubes could not perform the same storage ability as EDTA-K3 tube at "T0" time point. Plain tube had the largest drop among all types of blood collection tubes. Compared by pairwise t-test, EDTA-K3 tube could maintain intact PTH for 8 hours (p = 0.998) at room temperature and 24 hours (p = 0.053) in refrigeration. When comparing the total change limit (TCL = 18.8%), at room temperature, EDTA-K3 tube (7.0%), heparin tube (12.7%), coagulant tube (16.2%), and plain tube (17.6%) could maintain intact PTH for 8 hours, and GST can preserve PTH for 4 hours (18.2%). In refrigeration, EDTA-K3 tube could maintain PTH for 72 hours (7.5%) and heparin tube could maintain 24 hours (18.4%). The other three blood collection tubes could not preserve PTH in refrigeration (GST = 22.1%, coagulant tube = 20.3%, plain tube = 20.8%). CONCLUSIONS: PTH seems more stable in the EDTA-K3 tube than any other blood collection tubes and is followed next by the heparin anticoagulant tube. Plain tube and GST have faster degradation than other tubes and are not suggested to preserve intact PTH.

Intestinal gangrene secondary to congenital transmesenteric hernia in a child misdiagnosed with gastrointestinal bleeding: A case report.

BACKGROUND: Congenital transmesenteric hernia in children is a rare and potentially fatal form of internal abdominal hernia, and no specific clinical symptoms can be observed preoperatively. Therefore, this condition is not widely known among clinicians, and it is easily misdiagnosed, resulting in disastrous effects. CASE SUMMARY: This report presents the case of a 13-year-old boy with a chief complaint of abdominal pain and vomiting and a history of duodenal ulcer. The patient was misdiagnosed with gastrointestinal bleeding and treated conservatively at first. Then, the patient's symptoms were aggravated and he presented in a shock-like state. Computed tomography revealed a suspected internal hernia, extensive small intestinal obstruction, and massive effusion in the abdominal and pelvic cavity. Intraoperative exploration found a small mesenteric defect approximately 3.5 cm in diameter near the ileocecal valve, and there was about 1.8 m of herniated small intestine that was treated by resection and anastomosis. The patient recovered well and was followed for more than 5 years without developing short bowel syndrome. CONCLUSION: In this report, we review the pathogenesis, presentation, diagnosis, and treatment of congenital transmesenteric hernia in children.

ProGRP AS A NOVEL BIOMARKER FOR THE DIFFERENTIAL DIAGNOSIS OF MEDULLARY THYROID CARCINOMA IN PATIENTS WITH THYROID NODULES.

Objective: To investigate the release of progastrin-releasing peptide (ProGRP) in patients with thyroid nodules and the value of ProGRP in fine-needle aspirate washout fluid (FNA-ProGRP) in the differential diagnosis between medullary thyroid carcinoma (MTC) and non-MTC thyroid nodules. Methods: We investigated 2,446 healthy persons and 212 patients with 235 thyroid nodules. They were classified into healthy, nodular goiter, chronic thyroiditis, thyroid follicular neoplasm, papillary thyroid carcinoma, follicular thyroid carcinoma, and medullary thyroid carcinoma. The serum ProGRP and FNA-ProGRP were measured. Results: The serum ProGRP median concentration in MTC was 124.40 pg/mL, significantly higher than in other groups. The cutoff value of serum ProGRP was 68.30 pg/mL, leading to 53.85% sensitivity, 96.98% specificity, and 0.51 kappa value in MTC. The FNA-ProGRP median concentration in MTC nodules was 2,096.00 pg/mL, significantly higher than in other groups. A receiver operating characteristic analysis of MTC nodules and non-MTC nodules indicated that the cutoff value was 22.77 pg/mL, leading to 94.12% sensitivity, 98.27% specificity, and 0.85 kappa value. Conclusion: FNA-ProGRP measurement could be served as an ancillary method for the differential diagnosis between MTC and non-MTC thyroid nodules. Abbreviations: CEA = carcinoembryonic antigen; CT = calcitonin; FNAC = fine-needle aspiration cytology; FNA-CT = calcitonin in fine-needle aspirate washout fluid; FNA-ProGRP = ProGRP in fine-needle aspirate washout fluid; MTC = medullary thyroid carcinoma; ProGRP = progastrin-releasing peptide; SCLC = small-cell lung cancer; TM = tumor marker.

Expression and clinical value of gastrin-releasing peptide precursor in nephropathy and chronic kidney disease.

AIM: To explore whether serum pro-gastric releasing peptide (proGRP) is elevated in nephropathy patients and evaluate the diagnostic value of proGRP in chronic kidney disease (CKD) patients. METHODS: A total of 498 nephropathy patients and 170 healthy were selected in Zhongshan Hospital, Medical College of Xiamen University, from February 2016 to September 2017. The clinical data of the different groups including serum proGRP, CKD grading, and other serum and urine renal function biomarkers were analyzed by group comparison, correlation analysis and receiver operating characteristic curve. RESULTS: Serum proGRP levels were significantly higher in the acute kidney injury and CKD groups compared with the other groups of kidney disease patients (P < 0.01), and increased with CKD grading (P < 0.01). Serum proGRP was substantially correlated with serum creatinine (r = 0.637, P < 0.01) and cystain C (0.837, P < 0.01). Serum proGRP had moderate correlations with urine ß2-macroglobulin (ß2-m; r = 0.587, P < 0.01) and α1-macroglobulin (α1-m; r = 0.557, P < 0.01). There were fair associations of serum proGRP with albumin (r = 0.10, P = 0.067), 24 h proteinuria (24 h-TPU; r = 0.092, P = 0.099), urinary albumin/urocreatinine (uAlb/Cr; r = 0.29, P < 0.01) and urinary N-acetyl-ß-D-glucosidase (r = -0.142, P < 0.01). The sensitivity of proGRP was superior to that of simplified modification of diet in renal disease (MDRD) formula in diagnosing CKD I + II (81.25% vs 66.67%), CKD III (86.42% vs 74.36%) and CKD IV (71.19% vs 69.64%), while its specificity was inferior to that of simplified MDRD formula in diagnosing CKD I + II (37.65% vs 66.97%), CKD III (56.25% vs 86.67%) and CKD IV (75.31% vs 88.46%). CONCLUSION: Serum proGRP is elevated in acute renal injury and CKD patients and increases with CKD grading. Serum proGRP is mainly affected by glomerular filtration rate and could be used for CKD staging, although the overall diagnostic sufficiency is inferior to simplified MDRD formula.

Clinical Application of Pro-Gastrin-Releasing Peptide.

BACKGROUND: The diagnostic sensitivity and specificity of serum Pro-gastrin-releasing peptide (ProGRP) in benign or malignant diseases remains controversial. METHODS: The clinical data of 6,948 patients who were examined for ProGRP from February 2015 to January 2017 in the Zhongshan Hospital of the XiaMen University, were collected. The clinical data of the different groups were analyzed by statistical methods. RESULTS: The ProGRP reference levels were 68.50 pg/mL. The percentages of abnormal ProGRP levels were 23.87%, 26.14%, 22.87%, 84.56%, 16.1%, 15.59% and 43.75% for the pneumonia, cardiovascular disease, cerebral vascular disease, renal failure, small cell lung cancer (SCLC), colorectal cancer, and prostate cancer groups, respectively. The 95th percentile of the serum ProGRP concentration levels in the renal failure group was 364.22 pg/mL. The ProGRP cutoff levels for SCLC group were 162.90 pg/mL and the area under the receiver-operating characteristic curve (AUC) was 0.940 (95% CI, 0.892 to 0.987). The parameters sensitivity, specificity, PLR (positive likelihood ratio), NLR (negative likelihood) and Kappa value for the SCLC group with the exclusion of the renal failure group were 81.10% (95% CI: 38.19% - 89.71%), 99.51% (95% CI: 99.31% - 99.66%), 167.59 (95% CI: 116.06 - 241.99), 0.19 (95% CI: 0.11 - 0.32), and 0.6830 (Sig: 0.000), respectively. In the SCLC group, significantly higher concentrations of serum ProGRP of M1 were observed compared with M0. All esophageal neoplasms with high-level ProGRP were identified as small cell neoplasms. CONCLUSIONS: The ProGRP levels were increased in a variety of benign and malignant diseases and were considered a putative marker for SCLC with the exception of the renal failure group. It may be suggested that the high ProGRP levels originated from patients with neuroendocrine tumor.

Evaluation of a newly developed quantitative determination kit for tumor marker CA15-3 with chemiluminescent assay.

BACKGROUND: Tumor marker carbohydrate antigen 15-3 (CA15-3) is used as a biomarker to aid to diagnose and monitor the prognosis of breast cancer patients. A new quantitative determination kit for CA15-3 with chemiluminescent assay was developed by Xiamen InnoDx Biotech Co., Ltd, China. Therefore, we conducted the report to evaluate the performance of the kit. METHODS: According to the "Guiding principles on performance analysis of diagnostic reagents in vitro", the calibration curve, limit of detection, reportable range, accuracy, precision, anti-interference capability, cross-reaction and comparison by measuring EDTA plasma and serum were carried out. In addition, the kit was performed in parallel to electrochemiluminescence immunoassay kit (Roche) to analyze the correlation between the two kits. RESULTS: Regression equation of calibration curve of the kit was Y=0.7914X+4.1032 (R2 =.990). Limit of detection was 0.0347 U/mL. The reportable range was 0.5-2400 U/mL. Recovery ratio was 100.0%-104.8%. Coefficient of variations (CVs) of within-run and between-run were 4.8%-7.6% and 5.8%-7.4% respectively. No remarkable interferences (all Bias% were less than ±10%) were detected when samples contained hemoglobin ≤183.8 µmol/L, bilirubin ≤340 µmol/L, triglyceride ≤18.1 mmol/L, or rheumatoid factor ≤400 U/mL. No cross-reaction was present in the kit. Moreover, compared with the results from electrochemiluminescence immunoassay kit (Roche) in 345 serum samples, there was a satisfied correlation coefficient of 0.977 (P<.01), and the kit was simultaneously fit for the detection of EDTA plasma and serum samples. CONCLUSION: The new kit validated satisfactorily, and it can be used for detecting CA15-3 in clinical practice.

Surgical management of Hawkins type III talar neck fracture through the approach of medial malleolar osteotomy and mini-plate for fixation.

BACKGROUND: Fractures of the talar neck are relatively uncommon yet current interventions suffer from a high incidence of complications and poor functional outcomes. In the present study, we report a surgical treatment of Hawkins type III talar neck fracture through the approach of medial malleolar osteotomy and mini-plate for fixation and discuss the therapeutic effects after long-term follow-up. METHODS: From January 2010 to January 2015, 21 patients with 22 fractures were treated using this approach within days of sustaining the injury. Clinical and radiographic data were collected during regular post-operative follow-ups. Health-related quality of life factors were evaluated using visual analogue scale (VAS). Functional outcomes were determined according the Hawkins score and the Ankle-Hind foot Scale of the American Orthopedic Foot and Ankle Society (AOFAS). Present of complications such as arthritis, avascular necrosis (AVN), and malunion were evaluated using radiographs and magnetic resonance imaging (MRI). Anatomical parameters of injured and corresponding uninjured talus were measured and compared using digital three-dimensional (3D) computer model. RESULTS: The mean duration of surgery was 65.6 ± 9.7 min. The average blood loss volume of the patients was 29.1 ± 5.7 ml. All the patients except 1 were followed up 18 to 41 months (average 29.6 months). The average VAS score for these patients was 3.2 ± 1.1, and the mean Hawkins score was 11.4 ± 3.4 at the final follow-up visit. The average AOFAS score was 72.8 ± 17.3. Nine patients outcomes were rated as "excellent", 4 as "good", 4 as "fair," and 4 as "poor". No malunion, screw loosening, plate breakage, or other internal fixation failures were found at final follow-up. Long-term complications included: 1 case of malunion, 5 cases of complete AVN, 8 cases of partial AVN, 13 cases of talocrural arthritis, 14 cases of subtalar arthritis, and 3 cases of talonavicular arthritis. Secondary surgery was performed in 4 cases. The relevant average anatomical data of injured and uninjured talus show no significant difference. CONCLUSIONS: This surgical treatment we used here resulted in decreased soft tissue trauma, adequate exposure of talar neck, satisfactory performance of daily life activities, and quality of life following surgery and restoration of anatomy of injured talus. However, long-term complications such as arthritis and AVN are still commonly seen.

Clinical significance of combined testing of YKL-40 with CEA in Chinese colorectal cancer patients.

BACKGROUND: To investigate the practical value of individual and combined testing of plasma levels of YKL-40, CEA, and CA199 for auxiliary diagnosis and detection of recurrence of colorectal cancer. METHODS: ELISA and ECLIA were used to evaluate levels of YKL-40, CEA, and CA199 in 120 colorectal cancer patients (56 initial-diagnosis, 42 post-operative, and 22 recurrent cases). Forty-three patients with benign colorectal disease and 36 healthy patients were enrolled as controls. The relationship between YKL-40 and clinical indicators of tumor pathology was analyzed. The positive rate and diagnostic efficacy of single and combined YKL-40, CEA, and CA199 testing were assessed in patients with colorectal cancer. RESULTS: Plasma YKL-40 in the cancer group was significantly higher than in the benign control and healthy control group, and the mean values were 145.4 ng/mL, 107.7 ng/mL, and 51.3 ng/mL (p < 0.05), respectively. With 72 ng/mL as the diagnostic threshold, the sensitivity and specificity of YKL-40 in colorectal cancer diagnosis were found to be 73.2% and 66.7%, respectively. Early-stage colorectal cancer patients showed a YKL-40 positive rate (73%) significantly higher than those of CEA and CA199 (50% and 32%, respectively; p < 0.05). When YKL-40 testing was combined with CEA or CA199, the positive rate increased to 82.1% and 80.3%, respectively. Through ROC curve analysis of the post-operative recurrent group against the non-recurrent group, the areas under the curve for YKL-40, CEA, and CA199 were found to be 0.907, 0.714, and 0.759, respectively. Based on the Dukes classification, the mean YKL-40 value for stages A/B, C, and D were 120.1 ng/mL, 131.7 ng/mL, and 226.8 ng/mL (p = 0.008), respectively. The plasma YKL-40 level gradually increased as the disease progressed. Lower degrees of tumor differentiation were correlated with higher YKL-40 levels. The mean YKL-40 values of high, medium, and low tumor differentiation groups were 96.8 ng/mL, 127.5 ng/mL, and 225.7 ng/mL (p = 0.004), respectively. CONCLUSIONS: The benefits of using YKL-40 testing are higher than CEA and CA199 for the monitoring of colorectal cancer recurrence. Combined testing of both YKL-40 and CEA was found to be optimal for auxiliary diagnosis of colorectal cancer. Plasma YKL-40 was found to be suitable for auxiliary diagnosis of colorectal cancer.