Identification of a novel de novo PUF60 variant causing Verheij syndrome in a fetus.

Verheij syndrome (VRJS) is a craniofacial spliceosomopathy with a wide phenotypic spectrum. Haploinsufficiency of the poly-uridine binding splicing factor 60 gene (PUF60) and its loss-of-function (LOF) variants are involved in VRJS. We evaluated a human fetus with congenital heart defects and preaxial polydactyly. Clinical data were obtained from the medical record. Whole-exome sequencing (WES) was used to explore the potential genetic etiology, and the detected variant verified using Sanger sequencing. Functional studies were performed to validate the pathogenic effects of the variant. Using trio-WES, we identified a novel PUF60 variant (NM_078480.2; c.1678 T > A, p.*560Argext*204) in the pedigree. Bioinformatic analyses revealed that the variant is potentially pathogenic, and functional studies indicated that it leads to degradation of the elongated protein and subsequently PUF60 LOF, producing some VRJS phenotypes. These findings confirmed the pathogenicity of the variant. This study implicates PUF60 LOF in the etiopathogenesis of VRJS. It not only expands the PUF60 variant spectrum, and also provides a basis for genetic counseling and the diagnosis of VRJS. Although trio-WES is a well-established approach for identifying the genetic etiology of rare multisystemic conditions, functional studies could aid in verifying the pathogenicity of novel variants.

The clinical topography of peripherally inserted central catheter-related thrombosis in cancer patients: A prospective and longitudinal observational study based on ultrasound scans every two days.

AIMS: To delineate the clinical topography of peripherally inserted central catheter (PICC)-related thrombosis in cancer patients. BACKGROUND: Most of the clinical features of PICC-related thrombosis are based on a single follow-up, which is insufficient to reflect the full topography of a thrombosis. DESIGN: This is an observational study conducted at West China Hospital, according to the STROBE guidelines. METHODS: Cancer patients scheduled for PICC placement were potentially eligible; patients with contraindications to PICC placement or existing diseases affecting blood flow were excluded; and those who later withdrew or did not reply to our contact request during the follow-up period were eliminated from this study. Ultrasound was used to detect thrombosis from the insertion site, proximal insertion site, axillary vein to the subclavian vein once every two days for two weeks post insertion. The thrombosis and its involved venous segments, onset time and symptoms and signs were recorded. RESULTS: Among the 173 included patients, 126 (72.8 %) were identified as having thrombosis. Specifically, 113 and 126 patients were identified as having thrombosis within the first three days and the first week post insertion, respectively. In the 126 patients, thrombosis occurred at the insertion site (72.8 %) concurrently with thrombosis at the proximal insertion site (n = 120, 69.4 %), thrombosis in the axillary vein (n = 94, 54.3 %), and/or thrombosis in the subclavian vein (n = 41, 23.7 %). The log-rank test demonstrated that thrombosis in these four venous segments decreased significantly from the distal to the proximal central vein (log-rank test = 117.128, P < .001). Of 31 patients (17.9 %) who presented symptomatic thrombosis, only five patients experienced obvious swelling in the upper arm, and the other 26 patients exhibited atypical symptoms, such as soreness, tightness, numbness, tingling, or other discomforts in the palm, arm, armpit, and/or shoulder. In some thrombotic cases, ultrasonic assessment of PICC-related thrombosis did not parallel clinical symptoms and signs. CONCLUSION: PICC-related thrombosis is common and can occur very early post insertion in cancer patients, and most thromboses present atypical symptoms. More than half of the cases with thrombosis evaluated involve multiple venous segments, and the farther the venous segments are from the central vein, the higher the incidence of thrombosis tend to be and the earlier the onset time are. RELEVANCE TO CLINICAL PRACTICE: The results highlight the importance that medical staff pay particular attention to patients with catheters in the first week post insertion and be alert to thrombosis presenting atypical symptoms while keeping in mind that clinical symptoms and signs are not reliable for diagnosing thrombosis. CLINICAL REGESTRATION: Clinical Trials ChiCTR1900024890.

A novel PKHD1 splicing variant identified in a fetus with autosomal recessive polycystic kidney disease.

Objective: Variants of the polycystic kidney and hepatic disease 1 (PKHD1) gene are associated with autosomal recessive polycystic kidney disease (ARPKD). This study aimed to identify the genetic causes in a Chinese pedigree with ARPKD and design a minigene construct of the PKHD1 gene to investigate the impact of its variants on splicing. Methods: Umbilical cord samples from the proband and peripheral blood samples from his parents were collected, and genomic DNA was extracted for whole-exome sequencing (WES). Bioinformatic analysis was used to identify potential genetic causes, and Sanger sequencing confirmed the existence of variants within the pedigree. A minigene assay was performed to validate the effects of an intronic variant on mRNA splicing. Results: Two variants, c.9455del (p.N3152Tfs*10) and c.2408-13C>G, were identified in the PKHD1 gene (NM_138694.4) by WES; the latter has not been previously reported. In silico analysis predicted that this intronic variant is potentially pathogenic. Bioinformatic splice prediction tools revealed that the variant is likely to strongly impact splice site function. An in vitro minigene assay revealed that c.2408-13C>G can cause aberrant splicing, resulting in the retention of 12 bp of intron 23. Conclusion: A novel pathogenic variant of PKHD1, c.2408-13C>G, was found in a fetus with ARPKD, which enriches the variant spectrum of the PKHD1 gene and provides a basis for genetic counseling and the diagnosis of ARPKD. Minigenes are optimal to determine whether intron variants can cause aberrant splicing.

[Clinical Study of Peripherally Inserted Central Catheter-Related Thrombosis and Its Influence on the Blood Flow Status of the Inserted Veins in Cancer Patients].

Objective: To investigate the clinical features of peripherally inserted central catheter (PICC)-related thrombosis (PICCRT) within 2 weeks after PICC placement in cancer patients and its dynamic influence on the blood flow status of veins inserted with catheter, and to provide support for implementing thrombosis prevention and control measures. Methods: Between May 2019 and July 2020, patients who had solid tumors and who had PICC were prospectively enrolled at West China Hospital, Sichuan University. Scheduled color Doppler imaging was performed to examine the status of PICCRT formation at 8 points of time, with the first one conducted one day before the insertion of PICC and the other 7 completed within 2 weeks after the insertion of PICC. Then, based on whether patients had PICCRT, the patients were divided into two groups, a non-PICCRT group and a PICCRT group. The PICCRT group was further divided into two subgroups, an asymptomatic PICCRT group and a symptomatic PICCRT group, according to whether the patients had thrombosis-related symptoms and signs. Comparisons were made to study the incidence of PICCRT and the vascular diameter and the blood flow velocity in the veins inserted with catheters at different points of time in the patients of different groups. Results: Among 173 cancer patients in the cohort, 126 (72.8%) developed PICCRT, all of which occurred within 1 week after PICC insertion. There were 95 cases of asymptomatic PICCRT and 31 cases of symptomatic PICCRT. Before and after PICC insertion, the vascular diameter of both the asymptomatic and symptomatic PICCRT groups was significantly smaller than that of the non-PICCRT group and the blood flow velocity was significantly slower than that of the non-PICCRT group, with the difference continuing to increase with the prolongation of catheter indwelling time. Conclusion: Inserting catheters in veins with bigger vascular diameter and faster blood flow velocity may help reduce the incidence of PICCRT. The first week post catheter insertion is the key intervention period for the prevention of PICCRT.

The effect of systemic and local risk factors on triggering peripherally inserted central catheter-related thrombosis in cancer patients: A prospective cohort study based on ultrasound examination and structural equation modeling.

BACKGROUND: The negative synergistic effect of cancer and a peripherally inserted central catheter could significantly increase the incidence of thrombosis. Rather than identifying risk factors for peripherally inserted central catheter-related thrombosis, exploring the effect of these risk factors might be a promising method to improve the outcomes of thrombosis. OBJECTIVES: To analyze the effect of systemic and local risk factors on triggering peripherally inserted central catheter-related thrombosis in the first two weeks post-insertion in cancer patients. DESIGN: A prospective cohort study. SETTING: The study was conducted at a 4500-bed university-affiliated medical center in China. PARTICIPANTS: One hundred seventy-three cancer patients with peripherally inserted central catheters were included. METHODS: Peripherally inserted central catheter-related thrombosis was assessed using ultrasound at a series of timepoints, once every two days post-insertion. Data on age, body mass index, blood hypercoagulation, insertion attempts, catheter-to-vein ratio, and blood flow velocity were collected as risk factors. Descriptive statistics and structural equation modeling were used to describe the study samples and analyze the effects of systemic and local risk factors. RESULTS: Among the 173 cancer patients included, 126 (72.8%) patients were diagnosed with peripherally inserted central catheter-related thrombosis. Most thromboses (n = 118, 93.7%) were detected within five days, and 100% were detected within nine days post-insertion. Structural equation modeling analysis showed that local risk factors [catheter-to-vein ratio (standardized path coefficient = 0.32, p < 0.05) and blood flow velocity (standardized path coefficient = -0.35, p < 0.05)] had a greater effect than systemic factors [age (standardized path coefficient = 0.13, p < 0.05) and blood hypercoagulation (standardized path coefficient = 0.17, p < 0.05)] on triggering peripherally inserted central catheter-related thrombosis. CONCLUSION: Peripherally inserted central catheter-related thrombosis is quite common and can occur very early post-insertion in cancer patients. Among the common risk factors, local risk factors reflecting peripherally inserted central catheter technology itself had a greater effect than systemic risk factors reflecting predisposition to thrombosis. Clinical Registration: Clinical Trials ChiCTR1900024890.

Weight Change Trajectory in Patients With Locally Advanced Nasopharyngeal Carcinoma During the Peri-Radiation Therapy Period.

OBJECTIVES: To analyze the weight change trajectory in patients with locally advanced nasopharyngeal carcinoma (LANPC) before, during, and after radiation therapy for a time span of 40 weeks. SAMPLE & SETTING: 147 patients from a university-affiliated medical center in China were included. METHODS & VARIABLES: Body weight was measured weekly during intensive treatment and biweekly after radiation therapy. RESULTS: All 147 patients experienced critical weight loss during the peri-radiation therapy period. Overall, body weight remained basically unchanged during induction chemotherapy, followed by a sharp and severe decrease during radiation therapy. At 20 weeks after radiation therapy, body weight had increased only slightly from the lowest point. IMPLICATIONS FOR NURSING: A time-tailored intervention based on the weight change trajectory is necessary for patients with LANPC. According to the weight change trajectory, relevant interventions for maintaining body weight should be initiated as early as the second week of radiation therapy and no later than the fourth week of radiation therapy, and these interventions should continue for at least four weeks after radiation therapy.

Comparative meta-analysis of pyrocarbon and silicone for joint replacement surgery.

OBJECTIVE: To investigate the efficacy and complications that might be associated with pyrocarbon compared with silicone in patients undergoing joint replacement surgery. METHODS: The full-text papers about the clinical efficacy of pyrocarbon and silicone were retrieved from multiple databases. Review Manager version 5.0 was adopted for meta-analysis and analyses of sensitivity and bias. RESULTS: Ultimately, we studied 232 patients across eight studies that met the eligibility criteria. The meta-analysis suggested a significant difference between the pyrocarbon and silicone groups in terms of the Disabilities of the Arm, Shoulder, and Hand (DASH) score (standard mean difference (SMD) = 1.48; 95% CI [0.97, 1.99]; P = .009; P for Heterogeneity <0.00001; I = 63%); Visual Analogue Score (VAS) (SMD = 1.68; 95% CI [1.36, 1.99]; P < .00001; P for heterogeneity = 0.01; I = 61%), and the abnormal radiolucent line (RR = 6.66; 95% CI [3.19, 13.89]; P < .00001; P for heterogeneity = 0.87, I = 0%); and ossification development (RR = 0.90; 95% CI [0.56, 1.44], P = .66; P for heterogeneity = 0.94, I = 0%). CONCLUSION: This study showed that pyrocarbon might be an efficient material compared with silicone for joint replacement surgery, but resulted in poorer functional and pain outcomes compared with silicone.

[Expression and Clinical Significance of Serum LRG1 in Patients with Diffuse Large B-Cell Lymphoma before Treatment].

OBJECTIVE: To investigate the clinical significance of serum LRG1, LDH and ß2-MG in the recurrence of diffuse large B-cell lymphoma(DLBCL) after chemotherapy. METHODS: The serum levels of LRG1, LDH and ß2-MG of 80 patients with DLBCL were detected before treatment and followed up for these patients was performed. The cut-off value of non-recurrent survival was determined by ROC analysis. The correlation of three serum markers for predicting recurrence with prognostic factors of diffuse large B-cell lymphoma patients after treatment was analyzed by ROC curve. RESULTS: The serum levels of LDH, LRG1 and ß2-MG were higher in the groups with high tumor stageing, extranodal invasion and bone marrow involvement, respectively(P＜0.05). The optimal cut-off values for predicting recurrence risk determined by ROC analysis: LDH 402.37 U/L, LRG1 1.81 mg/L and ß2-MG 168.3 ng/L, respectively.COX multivariate regression analysis showed that serum LRG1 was an independent factor affecting the recurrence of diffuse large B-cell lymphoma(P＜0.05). CONCLUSION: The serum level of LRG1 may become a new biological marker to predict the recurrence risk of diffuse large B-cell lymphoma.

[Efficacy of Quercetin-sensitized Adriamycin for Treatment of Refractory Acute Leukemia].

OBJECTIVE: To investigate the chemotherapeutic efficency of quercetin sensitized adriamycin. METHOD: CCK-8 was used to detect the inhibitory effect of different doses of adriamycin, quercetin and quercetin combined with adriamycin on the proliferation of primary leukemia cells from patients with clinically refractory acute leukemia. Quercetin, adriamycin and their combination were used to treat non-irradiated T-ALL leukemia mice to observe the changes of survival curve and myocardial injury. RESULT: There was no significant difference in the inhibition rate of primary leukemia cell proliferation between the adriamycin concentration group (6, 0.6 and 0.06 µg/ml) and the adriamycin half-dose (3, 0.3 and 0.03 µg/ml) plus quercetin (0.25 mmol/L) group at three different time points (24, 48 and 72 hours). There was a significant difference in the inhibition rate of primary leukemia cell proliferation among the drug concentration groups, and the inhibition rate of primary leukemia cell proliferation was time-and concentration-dependent (r24h，a\c\e=0.995、r48h，a\c\e=1.000、r72h，a\c\e=0.984、r24h，b\d\f=0.993、r48h，b\d\f=0.999、r72h，b\d\f=0.960). In vivo experiments showed that the survival time of non-irradiated T-ALL leukemia mice treated with low-dose adriamycin combined with quercetin was not significantly prolonged compared with the high-dose adriamycin treatment group. The survival time of non-irradiated T-ALL leukemia mice treated with high dose of adriamycin and quercetin was significantly prolonged (P＜0.05). Compared with adriamycin group, the SOD activity in adriamycin combined with quercetin group increased significantly and the MDA content decreased. The results of transcriptome sequencing analysis showed that the expression of Ighv1-84 and Igkv6-14 in adriamycin combined quercetin group and quercetin group was lower than that in adriamycin group. The Ms4a1, Podx1, Mecom, Sh3bgr12, Bex4 and Tdrp expression in adriamycin combined quercetin group and adriamycin group were higher than that in quercetin group, while Crabp1 expression was lower. CONCLUSION: Quercetin can inhibit the proliferation of primary leukemia cells in a time-dependent manner. Quercetin combined with adriamycin inhibit the proliferation of primary leukemia cells significantly, and had synergistic and additive effects on the proliferation of primary leukemia cells, and the inhibiting effect of quercetin combined with adriamycin is concentration-and time-dependent. Quercetin combined with high-dose adriamycin can significantly prolong the survival time of non-irradiated T-ALL leukemia mice and reduce the myocardial damage caused by adriamycin.

Combination of quercetin and Adriamycin effectively suppresses the growth of refractory acute leukemia.

The present study aimed to investigate the effect of combined treatment with quercetin and Adriamycin (doxorubicin) on the development of refractory acute leukemia. Primary leukemic cells were isolated from patients with refractory drug-resistant acute leukemia. The Cell Counting Kit-8 assay was used to detect the proliferation of cells treated with a range of doses of Adriamycin, quercetin and a combination of the two drugs. Non-irradiated mice were used to establish a T cell acute lymphoblastic leukemia (T-ALL) model, which was subsequently treated with Adriamycin, quercetin and a combination of the two drugs. The survival time was recorded, and white and red blood cells and platelets in mouse peripheral blood were counted. Superoxide dismutase (SOD) activity and malondialdehyde (MDA) content of cardiac tissues were measured as indicators of oxidative stress and damage. Proliferation of primary leukemic cells was reduced by Adriamycin depending on the dose (0.06, 0.6 or 6 µg/ml) and treatment duration (24, 48 or 72 h) compared with the vehicle treated group. Co-treatment with quercetin achieved a similar suppression of leukemic cell proliferation when a lower dose of Adriamycin (0.03, 0.3 or 3 µg/ml) was administered for the same duration. The survival of non-irradiated mice with T-ALL was improved by co-treatment with a high dose of Adriamycin and quercetin compared with either treatment alone. Compared with treatment with Adriamycin alone, the combined treatment with Adriamycin and quercetin significantly enhanced the SOD activity and reduced the MDA content in the heart. Therefore, quercetin may enhance the effects of Adriamycin on refractory acute leukemia.

Ginsenoside Rb3 strengthens the hypoglycemic effect through AMPK for inhibition of hepatic gluconeogenesis.

Ginsenoside Rb3 is one of the major active components in protopanaxdiol type ginsenosides, and has demonstrated anti-diabetic activity. However, the mechanism of this action has yet to be elucidated. The present study investigated the effects of ginsenoside Rb3 on the AMP-activated protein kinase (AMPK) gluconeogenesis pathway. The present study involved the use of HepG2 cells and western blot analysis to systematically evaluate the effect of ginsenoside Rb3 on AMPK signaling proteins and key factors of gluconeogenesis [phosphoenolpyruvate carboxykinase (PEPCK), glucose-6-phosphatase, forkhead transcription factor 1 (FOXO1) and hepatic nuclear receptor 4α (HNF4α)]. The results indicated that 25 µM ginsenoside Rb3 significantly activated AMPK activity, increased the ratio of p-AMPK/total-AMPK, and had synergistic effects with the activator of AICAR on the activation of AMPK. Further analysis indicated that the expression of the transcription factor FOXO1 and HNF4α protein, two important factors in the pathway of HepG2 cell gluconeogenesis, was significantly suppressed by ginsenoside Rb3. PEPCK and G6Pase were subsequently inhibited, which led to the suppression of gluconeogenesis. These effects were partially blocked by the AMPK inhibitor, Compound C, which indicated that the inhibition effects of ginsenoside Rb3 on hepatic gluconeogenesis were predominantly due to the activation of the AMPK signaling pathway. These data suggested that ginsenoside Rb3 can suppress hepatic gluconeogenesis, at least partially through stimulation of AMPK activity.