Genetic analysis and in vitro validation of a case of Alport syndrome due to a splicing variant of COL4A5 gene / 中华医学遗传学杂志

OBJECTIVE@#To explore the genetic basis for a patient with Alport syndrome (AS) and confirm the existence of a splicing variant.@*METHODS@#An AS patient diagnosed at the Affiliated Hospital of Inner Mongolia Medical University on January 8, 2021 for significant proteinuria and occult hematuria was selected as the study subject. Clinical data was collected. Peripheral blood samples were collected for the extraction of genomic DNA. Whole exome sequencing and Sanger sequencing were carried out to identify potential genetic variants. An in vitro experiment was also conducted to verify the abnormal mRNA splicing. Bioinformatic software was used to analyze the conservation of amino acids of the variant sites and simulate the 3D structure of the variant collagen IV protein. Immunofluorescence and immunohistochemistry were carried out on renal tissues from the patient to confirm the presence of AS kidney injury.@*RESULTS@#The patient, a 21-year-old male, had a 24-hour urine protein of 3.53 g/24 h, which fulfilled the diagnostic criteria for proteinuria. His blood uric acid has also increased to 491 μmol/L. DNA sequencing revealed that he has harbored a c.835-9T>A splice variant of the COL4A5 gene, which was not found in either of his parents. In vitro experiment confirmed that the variant has removed 57 bp from the exon 15 of the mRNA of the COL4A5 gene. The deletion may cause loss of amino acid residues from positions 279 to 297, which in turn may affect the stability of the secondary structure of the α5 chain encoded by the COL4A5 gene. The amino acids are conserved across various species. The result of homology modeling indicated that the trimerization of Col-IV with the mutated α5 chain could be achieved, however, the 3D structure was severely distorted. The AS kidney damage was confirmed through immunofluorescence assays. Based on the guidelines from the American College of Medical Genetics and Genomics, the c.835-9T>A variant was classified as likely pathogenic (PVS1_Moderate+PS3_Moderate+PM2_Supporting+PS2+PP3+PP4).@*CONCLUSION@#The c.835-9T>A variant of the COL4A5 gene probably underlay the AS in this patient. In vitro experiment has confirmed the abnormal splicing caused by the variant. Histopathological examination of the kidney tissue has provided in vivo evidence for its pathogenicity. Above finding has expanded the mutational spectrum of the COL4A5 gene.

Risk of gestational diabetes recurrence and the development of type 2 diabetes among women with a history of gestational diabetes and risk factors: a study among 18 clinical centers in China / 中华医学杂志(英文版)

BACKGROUND@#Gestational diabetes mellitus (GDM) brings health issues for both mothers and offspring, and GDM prevention is as important as GDM management. It was shown that a history of GDM was significantly associated with a higher maternal risk for GDM recurrence. The incidence of GDM recurrence was unclear because of the incidence of second-child was low before 2016 in China. We aim to investigate the prevalence of GDM recurrence and its associated high-risk factors which may be useful for the prediction of GDM recurrence in China.@*METHODS@#A retrospective study was conducted which enrolled participants who underwent regular prenatal examination and delivered twice in the same hospital of 18 research centers. All participants were enrolled from January 2018 to October 2018, where they delivered the second baby during this period. A total of 6204 women were enrolled in this study, and 1002 women with a history of GDM were analyzed further. All participants enrolled in the study had an oral glucose tolerance test (OGTT) result at 24 to 28 weeks and were diagnosed as GDM in the first pregnancy according to the OGTT value (when any one of the following values is met or exceeded to the 75-g OGTT: 0 h [fasting], ≥5.10 mmol/L; 1 h, ≥10.00 mmol/L; and 2 h, ≥8.50 mmol/L). The prevalence of GDM recurrence and development of type 2 diabetes mellitus were calculated, and its related risk factors were analyzed.@*RESULTS@#In 6204 participants, there are 1002 women (1002/6204,16.15%) with a history of GDM and 5202 women (5202/6204, 83.85%) without a history of GDM. There are significant differences in age (32.43 ± 4.03 years vs. 33.00 ± 3.34 years vs. 32.19 ± 3.37 years, P  < 0.001), pregnancy interval (4.06 ± 1.44 years vs. 3.52 ± 1.43 years vs. 3.38 ± 1.35 years, P  = 0.004), prepregnancy body mass index (BMI) (27.40 ± 4.62 kg/m2vs. 23.50 ± 3.52 kg/m2vs. 22.55 ± 3.47 kg/m2, P < 0.001), history of delivered macrosomia (22.7% vs. 11.0% vs. 6.2%, P < 0.001) among the development of diabetes mellitus (DM), recurrence of GDM, and normal women. Moreover, it seems so important in the degree of abnormal glucose metabolism in the first pregnancy to the recurrence of GDM and the development of DM. There are significant differences in OGTT levels of the first pregnancy such as area under the curve of OGTT value (18.31 ± 1.90 mmol/L vs. 16.27 ± 1.93 mmol/L vs. 15.55 ± 1.92 mmol/L, P < 0.001), OGTT fasting value (5.43 ± 0.48 mmol/L vs. 5.16 ± 0.49 mmol/L vs. 5.02 ± 0.47 mmol/L, P < 0.001), OGTT 1-hour value (10.93 ± 1.34 mmol/L vs. 9.69 ± 1.53 mmol/L vs. 9.15 ± 1.58 mmol/L, P < 0.001), OGTT 2-hour value (9.30 ± 1.66 mmol/L vs. 8.01 ± 1.32 mmol/L vs. 7.79 ± 1.38 mmol/L, P < 0.001), incidence of impaired fasting glucose (IFG) (fasting plasma glucose ≥5.6 mmol/L) (31.3% vs. 14.6% vs. 8.8%, P < 0.001), and incidence of two or more abnormal OGTT values (68.8% vs. 39.7% vs. 23.9%, P < 0.001) among the three groups. Using multivariate analysis, the factors, such as age (1.07 [1.02-1.12], P = 0.006), prepregnancy BMI (1.07 [1.02, 1.12], P  = 0.003), and area under the curve of OGTT in the first pregnancy (1.14 [1.02, 1.26], P  = 0.02), have an effect on maternal GDM recurrence; the factors, such as age (1.28 [1.01-1.61], P  = 0.04), pre-pregnancy BMI (1.26 [1.04, 1.53], P = 0.02), and area under the curve of OGTT in the first pregnancy (1.65 [1.04, 2.62], P = 0.03), have an effect on maternal DM developed further.@*CONCLUSIONS@#The history of GDM was significantly associated with a higher maternal risk for GDM recurrence during follow-up after the first pregnancy. The associated risk factors for GDM recurrence or development of DM include age, high pre-pregnancy BMI, history of delivered macrosomia, the OGTT level in the first pregnancy, such as the high area under the curve of OGTT, IFG, and two or more abnormal OGTT values. To prevent GDM recurrence, women with a history of GDM should do the preconception counseling before preparing next pregnancy.

Effects of interpregnancy interval on pregnancy outcomes of subsequent pregnancy: a multicenter retrospective study / 中华妇产科杂志

Objective:To explore the effects of interpregnancy interval (IPI) on pregnancy outcomes of subsequent pregnancy.Methods:A multicenter retrospective study was conducted in 21 hospitals in China. Information of age, height, pre-pregnancy weight, IPI, history of diseases, complications of pregnancy, gestational age of delivery, delivery mode, and pregnancy outcomes of the participants were collected by consulting medical records of pregnant women who had two consecutive deliveries in the same hospital during 2011 to 2018. The participants were divided into 4 groups according to IPI:<18 months, 18-23 months, 24-59 months and ≥60 months. According to the WHO′s recommendation, with the IPI of 24-59 months group as a reference, to the effects of IPI on pregnancy outcomes of subsequent pregnancy were analyzed. Stratified analysis was further carried out based on age, history of gestational diabetes mellitus (GDM), macrosomia, and premature delivery, to explore the differences in the effects of IPI on pregnancy outcomes among women with different characteristics.Results:A total of 8 026 women were included in this study. There were 423, 623, 5 512 and 1 468 participants in <18 months group, 18-23 months group, 24-59 months group and ≥60 months group, respectively. (1) The age, pre-pregnancy body mass index (BMI), history of cesarean section, GDM, gestational hypertension and cesarean section delivery rate of <18 months group, 18-23 months group, 24-59 months group and ≥60 months group were gradually increased, and the differences were statistically significant ( P<0.05). (2) After adjusting for potential confounding factors, compared with women in the IPI of 24-59 months group, the risk of premature delivery, premature rupture of membranes, and oligohydramnios were increased by 42% ( OR=1.42, 95% CI: 1.07-1.88, P=0.015), 46% ( OR=1.46, 95% CI: 1.13-1.88, P=0.004), and 64% ( OR=1.64, 95% CI: 1.13-2.38, P=0.009) respectively for women in the IPI≥60 months group. No effects of IPI on other pregnancy outcomes were found in this study ( P>0.05). (3) After stratified by age and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months would significantly increase the risk of oligohydramnios for women with advanced age ( OR=2.87, 95% CI: 1.41-5.83, P=0.004); and <18 months could increase the risk of premature rupture of membranes for women under the age of 35 ( OR=1.59, 95% CI: 1.04-2.43, P=0.032). Both the risk of premature rupture of membranes ( OR=1.58, 95% CI: 1.18-2.13, P=0.002) and premature delivery ( OR=1.52, 95% CI: 1.07-2.17, P=0.020) were significantly increased in the IPI≥60 months group. After stratified by history of GDM and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months would lead to an increased risk of postpartum hemorrhage for women with a history of GDM ( OR=5.34, 95% CI: 1.45-19.70, P=0.012) and an increased risk of premature rupture of membranes for women without a history of GDM ( OR=1.44, 95% CI: 1.10-1.90, P=0.009). After stratified by history of macrosomia and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months could increase the proportion of cesarean section for women with a history of macrosomia ( OR=4.11, 95% CI: 1.18-14.27, P=0.026) and the risk of premature rupture of membranes for women without a history of macrosomia ( OR=1.46, 95% CI: 1.12-1.89, P=0.005). After stratified by history of premature delivery and adjusted for confounding factors, compared with women in the IPI of 24-59 months group, IPI≥60 months would significantly increase the risk of premature rupture of membranes for women without a history of premature delivery ( OR=1.47, 95% CI: 1.13-1.92, P=0.004). Conclusions:Both IPI≥60 months and <18 months would increase the risk of adverse pregnancy outcomes in the subsequent pregnancy. Healthcare education and consultation should be conducted for women of reproductive age to maintain an appropriate IPI when they plan to pregnant again, to reduce the risk of adverse pregnancy outcomes in the subsequent pregnancy.

Relationship between B-tupe natriuretic peptide and preeclampsia of hypertensive disorder complicating pregnancy as well as its significance / 中国综合临床

Objective To investigate and evaluate the correlation between brain natriuretic peptide (BNP) and gestational hypertension and preeclampsia of hypertensive disorder complicating pregnancy (HDCP).Methods Fifty cases with HDCP and 46 cases with mild and 83 cases with sever stage preeclampsia were selected as our subjects.And 33 cases with regular pregnancy and 31 with irregular pregnancy were served as control group.Plasma brain natriuretic peptide,urinary protein quantity(UBQ),24-hour urinary protein assay (UPA) were measured.The correlations of brain natriuretic peptide and UBQ,UPA,systolic pressure (SP),diastolic pressure (DP) were analyzed.Results The levels of brain natriuretic peptide in the group with gestational hypertension and mild,severe preeclampsia groups were (48.54± 18.27),(79.46± 32.18) and (292.24±213.08) ng/L,higher than that in normal pregnancy and non pregnant group ((27.84± 14.58) and (20.63± 8.28) n/L;F =49.583,P＜0.05).While no significant difference exists between normal pregnancy group and non pregnant group.Grouped on the median values (199) of brain natriuretic peptide of the severe preeclampsia group,the levels of 24-hour UPA,systolic pressure and diastolic pressure were (5.46±2.68) g,(174.55± 13.58) mmHg,(113.74±9.91) mmHg in patients with brain natriuretic peptide ≥ 199 ng/L(n=42),significant higher than those in patients with brain natriuretic peptide ＜ 199 ng/L(n =41;(4.34± 1.95)g,(165.31±11.12) mmHg,(106.05±8.02) mmHg;t=2.603,3.396,2.308;P=＞0.010,0.001,0.024).The levels of 24-hour UPA,systolic pressure and diastolic pressure of patients with brain natriuretic peptide ≥ 86ng/L(n=20) in mild preeclampsia were (1.68±0.27) g,(163.69±8.29) mmHg,(105.45±6.71) mmHg,significant higher than those in patients with brain natriuretic peptide ＜ 86 ng/L (n =26;(1.16 ± 0.31) g,(152.90±7.32) mmHg,(99.19 ± 5.25) mmHg;t =3.180,2.508,2.32;P =0.010,0.016,0.025).Brain natriuretic peptide was closely correlated with UPA,systolic pressure and diastolic pressure in hypertensive disorder complicating pregnancy (HDCP) (r =0.29,0.30;P ＜ 0.01).Brain natriuretic peptide was closely correlated with UPA systolic pressure and diastolic pressure in mild preeclampsia (r =0.39,0.37,0.40;P ＜0.01).And correlation efficacy of brain natriuretic peptide with UPA,systolic pressure and diastolic pressure were 0.44,0.42 and 0.53 (P＜0.01).Conclusion The level of brain natriuretic peptide is closely associated with the severity of gestational hypertension and preeclampsia of hypertensive disorder complicating pregnancy.Correlation of brain natriuretic peptide to the severity of gestational hypertension and preeclampsia is independent of urinary protein and hypertension.Brain natriuretic peptide is an important indicator for the severity of gestational hypertension and preeclampsia of hypertensive disorder complicating pregnancy.

The clinical application of CT perfusion in assessing the status of axillary lymph nodes in patients with breast cancer / 中华放射学杂志

Objective To evaluate the clinical application of CT perfusion in predicting the status of axillary lymph nodes in patients with breast cancer.Methods Fony-five patients with infiltrating breast cancers and 46 clinically palpable axillary lymph nodes underwent dynamic mtdti-slice spiral CT(MSCT).Semi-automatic calculation of perfusion parameters including blood flow(BF),blood volume(BV),mean transit time(MTT)and permeability surface(PS)of "target" lymph nodes and muscles in the same scan level were respectively meagured and analyzed.Nonparametric Mann-Whitney U test was used for the statistics.Results Forty-six "target" lymph nodes examined by CTP were metastasis in 32 cases and reactive hyperplastic lymph node inflammation in 14 cases at pathology.22 of 32 metastatic "target" nodes (68%)were sentinel lymph nodes(SLN).BF of CIP for inflammation and metastatic "target" nodes were (76.18±31.53)and(161.60±40.94)ml·100 mg-1·min-1,BV were(5.81±2.50)and(9.15±3.02)ml／100 mg.MTT were(6.80±1.55)and(5.50±1.84)s,PS were(25.82±4.62)and (25.96±7.47)ml·100 mg-1·min-1.There were significant correlations between the BF value of inflammation and metastatic "target" nodes(r=0.14,P<0.05)and there were no significant correlations among the BV,MTT and PS values of inflammation and metastatic "target" nodes(r=-0.03,0.05,0.07.P>0.05).Conelusion CTP can provide useful informafion for evaluating lymph node status.