Objective·To investigate the genetic etiology of a rare and complex case clinically suspected to be methylmalonic acidemia(MMA),but with negative whole exome sequencing(WES)results,using a multi-omics sequencing approach.Methods·DNA and RNA samples were extracted from the peripheral blood of the proband and both parents.Targeted MMA-related gene Panel sequencing and WES were first performed.Subsequently,RNA sequencing(RNA-seq)and whole genome sequencing(WGS)were conducted to comprehensively analyze the child's genetic variants,their origins and potential inheritance patterns.Results·No pathogenic variants associated with the patient's phenotype were identified through the MMA Panel or standard WES analysis.Extended analysis of WES suggested the possibility of uniparental disomy(UPD)of chromosome 4.WGS revealed a homozygous splice-site variant(c.-66+2T>C)in the non-coding region of the metabolism of cobalamin associated A(MMAA)gene.The variant was located in the 5'untranslated region(5'UTR),specifically at the second base downstream of the splice donor site of exon 1(reference sequence:NM_172250).In genomic coordinates(hg19),the variant was located at base 146540561 on chromosome 4(chr4:146540561).Sanger sequencing confirmed that the mother was heterozygous for this variant,while the father did not carry it.RNA-seq showed no detectable expression of the MMAA gene on chromosome 4 in the patient.This was further confirmed by reverse transcription real time quantitative PCR,indicating nearly absent mRNA expression,suggesting that the non-coding splice-site variant affected transcriptional expression.Conclusion·A homozygous splice-site variant(c.-66+2T>C)in the non-coding region of the MMAA gene—outside the coverage of WES—is likely the pathogenic cause in this case,presumably resulting from maternal UPD of chromosome 4.

Vitamin D is a fat-soluble vitamin primarily synthesized through skin exposure to ultraviolet B irradiation and dietary intake.Its biological effects are not limited to the regulation of calcium and phosphorus metabolism but also involve a variety of physiological functions such as immune modulation,anti-inflammation,and anti-tumor.In recent years,as research deepens,the role of the vitamin D signaling pathway in various diseases has been gradually revealed,and its regulatory mechanisms are complex and diverse.This paper systematically reviews the molecular mechanisms underlying the vitamin D signaling pathway,including the two-step hydroxylation activation process of vitamin D,the regulation of gene transcription mediated by the vitamin D receptor(VDR),the homeostatic regulation involving vitamin D-binding protein and metabolic enzymes such as 1α-hydroxylase and 24-hydroxylase,and interactions with other signaling pathways,including NF-κB,Wnt,and Hedgehog.This study highlights the role of vitamin D in various multi-system diseases such as inflammatory bowel disease,diabetes and its complications,obesity,cardiovascular disease,colorectal cancer,breast cancer,among others.The systematic cognitive framework for understanding the vitamin D signaling pathway was conducted,providing a theoretical basis for precision treatment strategies targeting VDR.

Vitamin D is a fat-soluble vitamin primarily synthesized through skin exposure to ultraviolet B irradiation and dietary intake.Its biological effects are not limited to the regulation of calcium and phosphorus metabolism but also involve a variety of physiological functions such as immune modulation,anti-inflammation,and anti-tumor.In recent years,as research deepens,the role of the vitamin D signaling pathway in various diseases has been gradually revealed,and its regulatory mechanisms are complex and diverse.This paper systematically reviews the molecular mechanisms underlying the vitamin D signaling pathway,including the two-step hydroxylation activation process of vitamin D,the regulation of gene transcription mediated by the vitamin D receptor(VDR),the homeostatic regulation involving vitamin D-binding protein and metabolic enzymes such as 1α-hydroxylase and 24-hydroxylase,and interactions with other signaling pathways,including NF-κB,Wnt,and Hedgehog.This study highlights the role of vitamin D in various multi-system diseases such as inflammatory bowel disease,diabetes and its complications,obesity,cardiovascular disease,colorectal cancer,breast cancer,among others.The systematic cognitive framework for understanding the vitamin D signaling pathway was conducted,providing a theoretical basis for precision treatment strategies targeting VDR.

Objective·To investigate the genetic etiology of a rare and complex case clinically suspected to be methylmalonic acidemia(MMA),but with negative whole exome sequencing(WES)results,using a multi-omics sequencing approach.Methods·DNA and RNA samples were extracted from the peripheral blood of the proband and both parents.Targeted MMA-related gene Panel sequencing and WES were first performed.Subsequently,RNA sequencing(RNA-seq)and whole genome sequencing(WGS)were conducted to comprehensively analyze the child's genetic variants,their origins and potential inheritance patterns.Results·No pathogenic variants associated with the patient's phenotype were identified through the MMA Panel or standard WES analysis.Extended analysis of WES suggested the possibility of uniparental disomy(UPD)of chromosome 4.WGS revealed a homozygous splice-site variant(c.-66+2T>C)in the non-coding region of the metabolism of cobalamin associated A(MMAA)gene.The variant was located in the 5'untranslated region(5'UTR),specifically at the second base downstream of the splice donor site of exon 1(reference sequence:NM_172250).In genomic coordinates(hg19),the variant was located at base 146540561 on chromosome 4(chr4:146540561).Sanger sequencing confirmed that the mother was heterozygous for this variant,while the father did not carry it.RNA-seq showed no detectable expression of the MMAA gene on chromosome 4 in the patient.This was further confirmed by reverse transcription real time quantitative PCR,indicating nearly absent mRNA expression,suggesting that the non-coding splice-site variant affected transcriptional expression.Conclusion·A homozygous splice-site variant(c.-66+2T>C)in the non-coding region of the MMAA gene—outside the coverage of WES—is likely the pathogenic cause in this case,presumably resulting from maternal UPD of chromosome 4.

Refeeding syndrome(RFS)has a high incidence among critically ill patients and significantly impacts the re-covery and prognosis of the patients.In this paper,we reviewed the literature on the risk factors and risk prediction models for RFS,finding the risk factors of RFS included patient-related,treatment-related factors and disease-related factors and the risk prediction models encompassed risk stratification model,risk score models and the Logistic regression models.It was concluded from the review that early assessment was crucial to preventing the occurrence of RFS.However,there was still a lack of reliable RFS risk prediction models with good predictive performance.It was found as well that it was crucial for the prevention of RFS to attach importance to nutritional and serological indicators and other factors.It was expected to be a necessity to conduct prospec-tive and multicenter studies to develop a risk prediction model for predicting RFS for ICU patients.Our review provides a refer-ence for early assessment and intervention for critically ill patients with RFS.

Objective To observe the value of intratumoral and peritumoral radiomics models combined with clinical and routine CT features for differentiating adenocarcinoma in situ(AIS)and microinvasive adenocarcinoma(MIA)of lung.Methods Totally 180 patients with isolated AIS and 180 with isolated MIA were retrospectively included,among them 160 AIS cases and 160 MIA cases were randomly selected into training set(n=320),while the other 20 AIS cases and 20 MIA cases were selected into test set(n=40).In training set,clinical and conventional CT features being statistically different between AIS and MIA were obtained to construct clinical model.Besides,radiomics features were extracted from intratumoral(CTi)ROI,intra-and peritumoral 2 mm(CTi+p2mm)ROI and intra-and peritumoral 4 mm(CTi+p4mm)ROI,and then CTi model,CTi+p2mm model and CTi+p4mm model for differentiating MIA and AIS were constructed.The optimal radiomics model for predicting MIA was selected using the area under the curve(AUC)of receiver operating characteristic(ROC)curve,and a combined model was built based on the optimal radiomics model combining with clinical and conventional CT features.The AUC,calibration and net benefit of the clinical model,the optimal radiomics model and the combined model were assessed.Results In training set,the larger nodular diameter,higher percentage of inhomogeneous density and ratio of nodules with vascular signals were observed in MIA compared with those in AIS(all P＜0.05).In test set,CTi+p2mm model had the highest efficacy(AUC=0.838)for differentiating MIA from AIS(P＜0.05),and the combined model had better efficacy(AUC=0.867,P＜0.05).The calibration of combined model was good,and the net benefit was high in 0.60-0.90 threshold probability range.Conclusion The radiomics model constructed with intratumoral and peritumoral 2 mm ROI based on plain CT was effective for differentiating MIA from AIS.Combining with clinical and routine CT features could furtherly improve differential diagnostic efficacy.

OBJECTIVE: To investigate the clinical features and genetic etiology of a case of Turner syndrome (TS) with rapidly progressive puberty. METHODS: A child who had presented at the Pediatric Endocrinology Clinic of the Shenzhen People's Hospital on January 19, 2022 was selected as the study subject. Clinical data of the child were collected. Peripheral blood sample of the child was subjected to chromosomal microarray analysis (CMA) and multiple ligation-dependent probe amplification (MLPA). Previous studies related to TS with rapidly progressive puberty were retrieved from the CNKI, Wanfang Data Knowledge Service Platform, Boku, CBMdisc and PubMed databases with Turner syndrome and rapidly progressive puberty as the keywords. The duration for literature retrieval was set from November 9, 2021 to May 31, 2022. The clinical characteristics and karyotypes of the children were summarized. RESULTS: The child was a 13-year-and-2-month-old female. She was found to have breast development at 9, short stature at 10, and menarche at 11. At 13, she was found to have a 46,X,i(X)(q10) karyotype. At the time of admission, she had a height of 143.5 cm (< P3), with 6 ~ 8 nevi over her face and right clavicle. She also had bilateral simian creases but no saddle nasal bridge, neck webbing, cubitus valgus, shield chest or widened breast distance. She had menstruated for over 2 years, and her bone age has reached 15.6 years. CMA revealed that she had a 58.06 Mb deletion in the Xp22.33p11.1 region and a 94.49 Mb duplication in the Xp11.1q28 region. MLPA has confirmed monosomy Xp and trisomy Xq. A total of 13 reports were retrieved from the CNKI, Wanfang Data Knowledge Service Platform, Boku, CBMdisc and PubMed databases, which had included 14 similar cases. Analysis of the 15 children suggested that their main clinical manifestations have included short stature and growth retardation, and their chromosomal karyotypes were mainly mosaicisms. CONCLUSION The main clinical manifestations of TS with rapidly progressive puberty are short stature and growth retardation. Deletion in the Xp22.33p11.1 and duplication in the Xp11.1q28 probably underlay the TS with rapid progression in this child, which has provided a reference for clinical diagnosis and genetic counselling for her.
Humans ; Female ; Adolescent ; Puberty ; Turner Syndrome/genetics* ; Chromosomes, Human, X ; Karyotyping

【Objective】 To study the safety, effectiveness and nursing of blood/fluid warmer during the process of plateletpheresis in winter. 【Methods】 The blood re-transfusion speed during plateletpheresis in winter and the time of blood passing through the blood return pipeline was counted. The vitro blood was heated with a blood/fluid warmer under different temperature settings, and the rising speed of blood temperature was recorded. The blood samples were tested for blood routine examination, free Hb and erythrocyte morphology at 0, 15 and 30 minutes. In the process of plateletpheresis in winter, the blood donors′ ear temperature and the skin temperature near the reinfusion needle in the observation group and the controls were measured, and the blood donors were observed for shivering, arm chills, pain or other discomfort. After the blood donation, the thermal comfort was evaluated. 【Results】 There was no difference in the results of routine blood test and plasma free Hb test of vitro blood after warming at 41℃, 42℃ and 43℃ for 0, 15 and 30 minutes (P>0.05), and no change in erythrocyte morphology was found. The skin temperature near the reinfusion needle (before vs. after the start of phlebotomy) was statistically different by applying blood/fluid warmer or not(P<0.05), and no difference in the temperature between the start and end of phlebotomy was observed in the observation group(P>0.05). The vitro blood heating experiment showed that when the room temperature was within 22~24℃, the blood retransfusion speed was (100-120) mL/min; after the application of blood/fluid warmer, the temperature of reinfusion blood could be raised from 27℃ to 33~37℃. The proportion of feeling comfortable and very comfortable and the score of thermal comfort in the blood donors who used the warmer were higher than those in the controls (P<0.05). When the temperature of the warmer was set above 38℃, the average score of thermal comfort of blood donors was above 8. 【Conclusion】 It is safe to apply the blood/fluid warmer during the plateletsapheresis in winter, which can significantly improve the comfort of blood donors and reflect the humanized service of blood stations, and is worth popularizing.

Objective:To investigate the relationship between tube voltage kV value for head and neck CT angiography and body weight.Methods:A total of 120 patients with suspected vascular disease of the head and neck who underwent CT angiography of the head and neck in Beihai People's Hospital from January 2020 to May 2022 were included in this study. Patients were divided into three groups according to different tube voltages: group A (tube voltage 120 kV, n = 45), group B (tube voltage 100 kV, n = 45) and group C (tube voltage 80 kV, n = 30). Patients in group A were divided into group A1 (< 70 kg, n = 15), group A2 (70-85 kg, n = 15) and group A3 (> 85 kg, n = 15) according to different body weights. Patients in group B were divided into group B1 (< 70 kg, n = 15), group B2 (70-85 kg, n = 15) and group B3 (> 85 kg, n = 15) according to different body weights. Patients in group C were divided into group C1 (< 70 kg, n = 15) and group C2 (70-85 kg, n = 15) according to different body weights. Group C3 was not used. The contrast medium used was Loversol. The CT value, image noise, signal-to-noise ratio, contrast to noise ratio, and effective radiation dose of arterial vessels in each group were measured. The images were subjectively evaluated by two physicians who had senior professional titles using a 5-point rating scale. Results:Subjective score of image quality was all ≥ 3 grade in each group. There was no significant difference in image quality rating between groups A1 and A2 and groups B1, B2, and C1. There was a remarkable difference in image quality rating between groups A3, B3, and C2 and the other groups. There was a significant difference in the CT value of blood vessels at four different levels between groups A1, B1 and C1 ( F = 76.82, 64.62, 98.79, 71.85, all P < 0.001). There was a significant difference in CT value of blood vessels at four different levels between groups A2, B2 and C2 ( F = 159.82, 112.33, 108.22, 135.18, all P < 0.001). There was a significant difference in CT value of blood vessels at four different levels between groups A3 and B3 ( t = 4.40, 4.27, 3.91, 3.59, all P < 0.05). In groups B3 and C2, the image noise was remarkably increased, signal to noise ratio and contrast to noise ratio were remarkably decreased compared with those in the other groups. The effective radiation dose of arterial vessels in group B1 was 47% lower than that in group A1 and the effective radiation dose of arterial vessels in group C1 was 73% lower than that in group A1 ( F = 116.18, P < 0.001). The effective radiation dose of arterial vessels in group B2 was 49% lower than that in group A2, and the effective radiation dose of arterial vessels in group C2 was 66% lower than that in group A2 ( H = 35.40, P < 0.001). The effective radiation dose in group B3 was 35% lower than that in group A3 ( t = 3.59, P < 0.05). Conclusion:In CT angiography of the head and neck, the selection of tube voltage kV value is related to body weight. Tube voltage 80 kV is suitable for patients with a body weight < 70 kg, tube voltage 100 kV for patients with a body weight of 70-85 kg, and tube voltage 120 kV for patients with a body weight > 85 kg. These tube voltages can decrease effective radiation dose and ensure image quality, meeting the requirement for clinical diagnosis.

Lnc-HUR1 is an HBV-related long non-coding RNA, which can promote the proliferation of hepatoma cells and the occurrence and development of liver cancer. In this study we explored the effect of lnc-HUR1 on the apoptosis of hepatocellular carcinoma cells by taking the approach of immunoblotting, quantitative real time PCR, luciferase reporter assay, chromatin immunoprecipitation (ChIP) and flow cytometry. We found that overexpression of lnc-HUR1 significantly reduced the activity of caspase3/7 and the cleavage of PARP-1, while knocking down of lnc-HUR1 significantly increased the activity of caspase3/7 and promoted the cleavage of PARP-1 in HepG2 cells treated with TGF-β, pentafluorouracil or staurosporine. Consistently, the data from Annexin-V/PI staining showed that overexpression of lnc-HUR1 inhibited apoptosis, while knockdown of lnc-HUR1 promoted apoptosis. Moreover, overexpression of lnc-HUR1 up-regulated the apoptosis inhibitor Bcl-2 and down-regulated the pro-apoptotic factor BAX at both RNA and protein levels. In the CCL4-induced acute liver injury mice model, the expression of Bcl-2 in the liver tissue of lnc-HUR1 transgenic mice was higher than that of the control mice. The data from ChIP assay indicated that lnc-HUR1 reduced the enrichment of p53 on Bcl-2 and BAX promoters. All these results indicated that lnc-HUR1 inhibited the apoptosis by promoting the expression of apoptosis inhibitor Bcl-2 and inhibiting the expression of apoptosis promoting factor BAX. Further studies showed that lnc-HUR1 regulated the transcription of Bcl-2 and BAX in HCT116 cells, but had no effect on the expression of Bcl-2 and BAX in HCT116 p53^-/- cells, indicating that lnc-HUR1 regulates the transcription of Bcl-2 and BAX dependent upon the activity of p53. In conclusion, HBV upregulated lnc-HUR1 can inhibit the apoptosis of hepatoma cells. Lnc-HUR1 inhibits apoptosis by inhibiting the transcriptional activity of p53. These results suggest that lnc-HUR1 plays an important role in the occurrence and development of HBV-related hepatocellular carcinoma.
Animals ; Annexins/pharmacology* ; Apoptosis ; Carcinoma, Hepatocellular/genetics* ; Cell Proliferation ; Hep G2 Cells ; Hepatitis B virus/metabolism* ; Humans ; Liver Neoplasms/genetics* ; Mice ; Poly(ADP-ribose) Polymerase Inhibitors/pharmacology* ; Proto-Oncogene Proteins c-bcl-2/pharmacology* ; RNA, Long Noncoding/metabolism* ; Staurosporine/pharmacology* ; Transforming Growth Factor beta/pharmacology* ; Tumor Suppressor Protein p53/pharmacology* ; bcl-2-Associated X Protein/pharmacology*