Diabetic kidney disease (DKD) is a common microvascular complication of diabetes. "Internal heat leading to zheng" is the core pathogenesis of DKD, while "glucose toxicity" is transformed from subtle substances through "internal heat" and the cementation of various pathological products, which is pivotal to the transformation of diabetes to DKD. "Glucose toxicity" is characterized by deep and widespread heat, caused by various pathological factors, and its sticky nature makes it difficult to resolve, which can cause severe damage to the kidney collaterals. In the early stage of "glucose toxicity", it is yang pathogen, which can be transformed into yin pathogen in the later stage with disease progression. In clinical practice, treatment should be based on disease staging, with attention on grasping the pathogenesis of "internal heat leading to zheng" and identifying the nature of "glucose toxicity". During the diabetic period, clearing heat is the primary method, often using modified Yueju Pill and Dachaihu Decoction. In the early stage of DKD, treatment primarily focuses on clearing and penetrating latent heat to treat DKD, aiming to prevent toxic heat from transitioning from qi to blood. The approach emphasizes clearing heat and re-penetrating, detoxification, and re-clearing, often using a self-made modified Qingre Xiaozheng Decoction. In the middle and late stages of DKD, the focus shifts to clearing heat, eliminating zheng, strengthening vital qi, and dispelling turbidity, with commonly used treatments including the self-made modified Xiezhuo Xiaozheng Formula, Jingui Shenqi Pill, and Zhenwu Decoction.

Objective:To screen key genes of renal clear cell carcinoma based on bioinformatics methods, identify possible microRNA (miRNA)-mRNA action axis, and explore the expression of related genes in clear cell renal cell carcinoma tissues and cells.Methods:Gene expression profiles of GSE40435 and GSE71302 datasets were obtained from the Gene Expression Omnibus (GEO) database. TCGA-KIRC datasets were obtained from The Cancer Genome Atlas (TCGA) database. R software was used to identify the differentially expressed mRNA and miRNA, and the functional enrichment analysis was performed. STRING database and Cytoscape software were used to perform the protein interaction analysis. The prognosis-related differentially expressed miRNA was evaluated by the Oncomir database. The potential targeted genes regulated by miRNA were determined by using TargetScan and miRDB targeted gene prediction tools. The tissue samples and clinicopathological features of 34 patients with clear cell renal cell carcinoma in the First Hospital of Shanxi Medical University from June to December 2021 were collected, and normal renal cell line 293T and clear cell renal cell carcinoma cell line 786O were selected. The real-time fluorescence quantitative polymerase chain reaction (qRT-PCR), was used to detect the relative expression of genes; Western blotting and immunohistochemical staining were used to detect the expression levels of the targeted proteins. The dual luciferase reporter gene assay was carried out to verify the targeting relationship between genes.Results:A total of 1 351 differentially expressed mRNA and 50 differentially expressed miRNA were screened and identified. The result of functional enrichment analysis suggested that the fatty acid metabolism pathway and xenobiotic metabolism pathway were suppressed in clear cell renal cell carcinoma, while the apoptosis and immune response pathways were activated. Protein interaction analysis suggested that the signal transduction and protein ubiquitination pathways might play a key role in clear cell renal cell carcinoma. The screening results showed that miRNA-224-5p (miR-224-5p) was most closely associated with clear cell renal cell carcinoma progression and was highly expressed in tumor tissues, and its prognosis-related target gene was NEDD4L. The relative expression of NEDD4L mRNA in clear cell renal cell carcinoma tissues and paraneoplastic tissues were 0.138±0.103 and 1.000±0.026 ( t = 46.23, P < 0.05), and the relative expression of miR-224-5p was 1.000±0.043 and 0.129±0.108 ( t = 45.28, P < 0.05). The differences of NEDD4L mRNA and miR-224-5p expressions in different grades and stages of clear cell renal cell carcinoma tissues were statistically significant (all P < 0.05). The expression of NEDD4L protein was decreased in clear cell renal cell carcinoma. The relative expression of NEDD4L gene in 293T and 786O cells were 1.000±0.125 and 0.210±0.044 ( t = 17.52, P < 0.05); the relative expressions of miR-224-5p gene were 0.209±0.049 and 1.000±0.234 ( t = 10.61, P < 0.05). The relative expressions of NEDD4L mRNA in miRNA mimic group and negative control group were 0.236±0.062 and 1.000±0.024, and the difference was statistically significant ( t = 43.56, P < 0.05). NEDD4L protein expression was reduced in the miRNA mimic group. Dual luciferase reporter gene assay suggested that NEDD4L was a direct target gene of miR-224-5p. Conclusions:In clear cell renal cell carcinoma, miR-224-5p targets and regulates NEDD4L expression, and this mechanism may be related to carcinogenesis and progression of clear cell renal cell carcinoma.

Objective:To investigate the value of a cuproptosis-related differential long non-coding RNA (lncRNA) scoring formula related to the prognosis of clear cell renal cell carcinoma (ccRCC) patients in the clinical diagnosis, prognosis prediction and treatment options based on bioinformatics.Methods:Gene matrix and clinical data of ccRCC patients were obtained from the Cancer Genome Atlas (TCGA) database (update to 29 March, 2022). The expression data of 539 ccRCC tissues and 72 paracancerous normal tissues were collected from gene matrix; the data of 530 ccRCC were collected from clinical data. Pearson correlation analysis, Wilcoxon signed rank test and univariate Cox proportional risk model were used to analyze the screened cuproptosis-related differential lncRNA related to the prognosis. R software was used to randomly divide 530 ccRCC patients with survival data into training set (266 cases) and validation set (264 cases) according to approximate 1∶1 ratio. LASSO regression analysis was used to construct a cuproptosis-related differential lncRNA scoring formula and cross-validation was performed. Receiver operating characteristic (ROC) curve analysis was used to evaluate the specificity and sensitivity of cuproptosis-related differential lncRNA scoring formula, and the area of the curve (AUC) was calculated. According to the median risk value, all patients were divided into the low-risk group and high-risk group; Kaplan-Meier method was used to analyze the difference in the overall survival (OS) of patients in the low-risk group and high-risk group. T test was used to detect the differences in the risk value of patients with different clinicopathological characteristics. R package rms was used to construct the nomogram for predicting 1-year, 3-year, 5-year OS rates of ccRCC patients, R package pRRophetic was used to predict the half-inhibitory concentration ( IC50) of common targeted drugs such as sorafenib and sunitinib in clinical treatment of ccRCC patients, and IC50 value of patients in low-risk group and high-risk group was compared by using Wilcoxon signed rank test. Tissue samples of 20 ccRCC patients who underwent radical nephrectomy and were diagnosed with pathology and the matched paracancerous normal tissues were collected from the First Hospital of Shanxi Medical University between June 2021 and December 2021. Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of key lncRNA in ccRCC tissues. Results:Based on the expression matrix of 10 cuproptosis genes (FDX1, LIAS, LIPT1, DLD, DLAT, PDHA1, PDHB, MTF1, GLS, CDKN2A) of ccRCC patients in TCGA database, 153 cuproptosis-related differential lncRNA related to the prognosis were identified. According to LASSO regression analysis, a scoring formula of 4 cuproptosis-related differential lncRNA related to the prognosis was obtained, risk value was calculated as 0.020×AC015912.3+0.011×AC026401.3+0.063×AC103706.1+(-0.076)×EPB41L4A-DT. All patients were divided into high-risk group (≥0.76) and low-risk group (<0.76) based on the median value (0.76). ROC curve analysis showed that the scoring formula had good prediction accuracy in 1-year, 3-year, 5-year OS rates. In training set, validation set, the total cohort, the OS of patients in the high-risk group was worse than that in the low-risk group (all P < 0.001). The age, pathological degree, tumor staging, risk value calculated by cuproptosis-related differential lncRNA were independent influencing factors of OS (all P < 0.001). There were statistically significant differences in the risk value calculated by cuproptosis-related differential lncRNA scoring formula among patients with different pathological degree, tumor staging, T staging, N staging, M staging (all P < 0.01), while there were no statistically significant differences among patients with different gender and age (all P > 0.05). The established nomogram had good prediction accuracy in the 1-year, 3-year, 5-year OS rates. Sunitinib and sirolimus showed higher sensitivity in the high-risk group; axitinib, sorafenib and pazopanib showed higher sensitivity in the low-risk group. qRT-PCR results showed that relative expression level of AC015912.3 in ccRCC tissues was up-regulated compared with paracancerous tissues (1.00±0.04 vs. 0.68±0.24, t = 6.37, P < 0.01); the relative expression level of AC026401.3 in ccRCC tissues was up-regulated compared with paracancerous tissues (1.00±0.05 vs. 0.64±0.22, t = 7.29, P < 0.01); the relative expression level of AC103706.1 in ccRCC tissues was up-regulated compared with paracancerous tissues (1.00±0.04 vs. 0.64±0.21, t = 7.49, P < 0.01); the relative expression level of EPB41L4A-DT in ccRCC tissues was up-regulated compared with paracancerous tissues (1.00±0.06 vs. 0.73±0.10, t = 10.68, P < 0.01). Conclusions:Cuproptosis-related differential lncRNA scoring formula based on TCGA database can be used as a new marker for clinical diagnosis and prognosis prediction of ccRCC patients, which can help guide the clinical drug treatment of patients and facilitate accurate diagnosis and treatment.

Objective:To explore the effects of kangaroo nursing after cesarean section on the paternal role adaptation and subjective well-being of first-time fathers.Methods:In this randomized controlled trial, a total of 160 first-time fathers of newborns delivered by cesarean section from May 2021 to May 2022 in the First Affiliated Hospital of Wenzhou Medical University were continuously selected as research objects and randomly divided into experimental group (80 cases) and control group (80 cases) by random number table method. In the control group, the nursing staff carried out routine obstetric care after the birth of the newborns, the newborns were swaddled to keep warm and transferred to the ward, the fathers took care of them by the side; and the fathers were given postnatal routine education and guidance once a day, including newborn bathing, touching and umbilical buttock care, maternal breast care, breastfeeding, diet, medication and discharge precautions etc. The experimental group were given kangaroo nursing on care basis of control group: the father took a comfortable position that half leaned on the recliner with his shirt unbuttoned, the naked newborn was placed on the father′s chest at an angle of approximately 60 degree on the head up and feet down for maximum skin to skin contact with the help of researchers. The newborn′s back was covered with a blanket and wrapped in the father′s clothes, soft music was playing throughout the whole process, meanwhile, researchers encouraged interactions through verbal communications and touching between the father and the newborn. After newborns returned to the ward, they were given kangaroo nursing immediately by the fathers for the first time, and then twice a day in the same way, lasting for 30-40 min every time, and a total of 6 times of kangaroo nursing were completed during in first 3 days after birth. The average hospitalization period of both groups was 5 days. The father-role adjustment scale and subjective well-being scale were asseessed in the fathers, the independent sample t test was used to compare differences of father-role adjustment level and subjective well-being at the time of discharge between the two groups, the Pearson correlation analysis was used to evaluate the correlation between the two variables. Results:The total score of paternal role adaptation, and the three dimensions scores including role identity, parent-child attachment establishment and nursing behavior implementation in the experimental group were all significantly higher than those in the control group (83.91±7.90 vs 78.79±9.44, 28.94±2.64 vs 27.85±3.36, 28.39±3.15 vs 26.46±3.52, 26.59±4.26 vs 24.48±3.81) ( t=3.725, 2.276, 3.647, 3.309, all P<0.05). The total score of subjective well-being, and the three dimensions scores including health worry, energy, melancholy or pleasant mood in the experimental were all significantly higher than those in the control group (88.34±6.38 vs 83.43±7.26, 7.70±1.77 vs 6.95±1.46, 20.03±3.15 vs 18.76±2.94, 19.41±2.01 vs 17.99±2.68) ( t=4.546, 2.928, 2.625, 3.805, all P<0.05), and were all significantly higher than those before the intervention (74.83±5.84, 6.11±2.22, 16.98±2.68, 15.18±2.13) ( t=13.069, 5.207, 6.261, 13.332, all P<0.05). There was a positive correlation between fathers′ subjective well-being and paternal role adaptation ( r=0.614, P<0.001). Conclusions:The father-centered kangaroo nursing after cesarean section can improve postpartum paternal role adaptation and subjective well-being of first-time fathers. There is a positive correlation between the subjective well-being and the paternal role adaptation level.

Objective:To evaluate the safety and effectiveness of the model repair strategy for preputial defect in hypospadias surgery.Methods:From February 2017 to December 2022, 59 children in our hospital with an average age of (6.9±3.2) years were retrospectively analyzed. All of them were children with multiple failed hypospadias operations. According to the condition of penile scrotum skin, different methods of penile skin reconstruction were selected. Among them, 20 patients underwent penile skin flap reconstruction with simple foreskin and multiple small incision reduction, 22 patients with penile and scrotal transposition underwent penile lateral scrotum flap to complete ventral penis coverage, and 12 patients underwent middle scrotum flap with pedicled flap to cover ventral urethra. The penis was covered by inferior epigastric artery perforator flap in 5 patients.Results:The wound healed completely one week after the operation and no obvious scar formation was observed. The ventral flap of the penis was covered by the lateral scrotal flap of the penis, and no flap necrosis and wound infection were observed. The ventral urethra was covered with pedicled skin flap in the middle scrotal suture. The flap survived without scrotal hematoma. The inferior epigastric artery perforator flap was used to cover the patients with penile defect, all the flaps survived, and no complications such as wound infection and abdominal hernia occurred. No urethral fistula or urethral diverticulum was reported in all the patients. Urethral stricture occurred in 1 case of scrotal suture pedicled flap group and 1 case of lateral penis scrotal flap group, which was cured after expansion. One case with small incision and one case with lateral scrotal skin were cured after secondary repair by Mathieu method.Conclusions:For children with a history of multiple hypospadias operations, coverage of ventral skin defects of the penis is essential to reduce complications and obtain good appearance. This study summarized four effective strategies for covering penile defect from simple to complex. The small incision was used to reduce the expansion of all flaps. The lateral penile scrotal flap is more suitable for patients with penile scrotal transposition. The pedicled flap of scrotal suture and inferior superficial artery perforator flap are suitable for the repair of larger defect area.

Objective:This study aimed to explore the feasibility and clinical value of monitoring the progression of early kidney injury in type 2 diabetic patients by assessment of the urinary C-terminal agrin fragment (uCAF) with enzymatic chemiluminescence immunoassay.Methods:A total of 251 patients with type 2 diabetes, who attended the Second Affiliated Hospital of Wenzhou Medical University from October 2018 to March 2020, were included in this retrospective analysis. One hundred and fifty-six participants undergoing health check-up at the Second Affiliated Hospital of Zhejiang University School of Medicine in February 2021 served as controls. Basic clinical information, glycosylated hemoglobin type A 1c and serum creatinine values were recorded, and urine specimens were collected for urinary creatinine, urinary α 1 microglobulin(uα 1M), urinary immunoglobulin G (uIgG), urinary albumin, urinary N-Acetyl-B-D-glycosaminidase (uNAG) and uCAF measurements. Based on the estimated glomerular filtration rate (eGFR), 251 patients were classified into G1~G5 stage groups with 116, 22, 28, 55 and 30 patients in each group. One hundred and sixty-six patients with early diabetic kidney disease (stage G1-G3) were divided into subgroups A1 (79), A2 (48) and A3 (39) according to the urinary albumin/creatinine ratio (UACR), the uα1M levels were divided into uα1M subgroup 1 (83 cases), uα1M subgroup 2 (42 cases), and uα1M subgroup 3 (41 cases), and uIgG subgroup 1 (83 cases), uIgG subgroup 2 (42 cases), and uIgG subgroup 3 (41 cases) according to uIgG levels. The Spearman method was used to analyze the correlation between uCAF levels and eGFR, UACR, uα1M and uIgG levels. Results:(1) The linear range of the uCAF detected by enzymatic chemiluminescence immunoassay was 3.97-2 000.00 ng/ml, with a detection limit of 2.28 ng/ml, intra-batch coefficients of variation of 1.15% and 1.57%, inter-batch coefficients of variation of 1.63% and 5.78%, and a biological reference interval of <95.35 μg/g Cr. (2) The uCAF level and positive rate (UACR≥30 mg/g) increased with the decrease of eGFR from G1-G3, uCAF level was negatively correlated with eGFR value ( r=-0.543, P<0.000 1), and the positive rate increased from 24.14% (28/116) to 85.71% (24/28) from G1-G3. The uCAF level and positivity rate decreased with the decrease of eGFR from G4 to G5. uCAF level was positively correlated with eGFR value ( r=0.495, P<0.001), and the positivity rate decreased from 30.91% (17/55) to 23.33% (7/30) from G4 to G5. (3) In patients with early diabetic kidney disease, uCAF levels and positivity rates increased gradually with the increase of UACR. uCAF levels were positively correlated with UACR values ( r=0.602, P<0.001), and the uCAF positivity rate reached 21.52% (17/79) in the A1 subgroup. (4) uCAF level was positively correlated with uα1M and uIgG levels in patients with early diabetic kidney disease ( r=0.757, 0.596, both P<0.001). Conclusion:Analytical performance of enzyme chemiluminescence immunoassay for the detection of CAF is satisfactory and could be used a biomarker for monitoring damage and progression of early diabetic kidney disease in patients with type 2 diabetes.

Respiratory viruses can cause a variety of serious respiratory infections and diseases of tissues and organs outside the respiratory tract, raising a potentially severe threat to the society.Virus replication and survival rely on the internal mechanism of host cells, and the latter also produce a variety of restriction factors that target viral invasion, genome transcription and replication, and assembly and release to block viral infection.Herein, this study reviewed the research progress of the antiviral effects of the host restriction factors of common respiratory viruses and their underlying mechanisms.

OBJECTIVETo use next generation sequencing (NGS) to identify unknown abnormality of chromosome 9 in a fetus and explore its mechanism.

METHODSA pregnant woman with abnormal fetal ultrasound finding underwent amniocentesis for G-banded chromosomal analysis. Karyotyping was also performed on peripheral blood samples derived from its parents. Fetal blood sample was obtained for NGS testing to identify abnormality unrecognized by karyotyping.

RESULTSAnalysis of amniocytes has revealed a 46,XX,der(9)(?::p21 to qter) karyotype, while both parents had a normal karyotype. NGS analysis of the fetus revealed a 20.67 Mb duplication (4 454 279-25 126 275) at 9p21.3p24.2, which overlapped with that of the 9p duplication syndrome, and a 4.43 Mb deletion at 9p24.2p24.3 (10 001-4 442 364), which partially overlapped with that of 9p deletion syndrome and 46,XY sex reversal 4 region. Comparison of the sequencing data with reference genome database indicated direct duplication of 9p21.3p24.2, which was also supported by review of the morphology of chromosome 9p. Therefore, the karyotype of the fetus was verified to be 46,XX,der(9) dir dup(9)(p21.3p24.2), del(9)(p24.2p24.3).

CONCLUSIONCombined G-banded karyotyping and NGS can identify dir dup del(9p) with accuracy. Delineation of the mechanism of dir dup del(9p) and its genotype-phenotype correlation may facilitate genetic counseling and estimation of recurrence risk.

Adult ; Chromosome Banding ; Chromosome Deletion ; Chromosomes, Human, Pair 9 ; genetics ; Female ; Fetal Diseases ; diagnosis ; genetics ; Humans ; Male ; Pregnancy ; Prenatal Diagnosis ; Trisomy ; genetics

OBJECTIVETo analyze a case with Angelman syndrome (AS) using single nucleotide polymorphism array (SNP array) and explore its genotype-phenotype correlation.

METHODSG-banded karyotyping and SNP array were performed on a child featuring congenital malformations, intellectual disability and developmental delay. Mendelian error checking based on the SNP information was used to delineate the parental origin of detected abnormality. Result of the SNP array was validated with fluorescence in situ hybridization (FISH).

RESULTSThe SNP array has detected a 6.053 Mb deletion at 15q11.2q13.1 (22,770,421- 28,823,722) which overlapped with the critical region of AS (type 1). The parents of the child showed no abnormal results for G-banded karyotyping, SNP array and FISH analysis, indicating a de novo origin of the deletion. Mendelian error checking based on the SNP information suggested that the 15q11.2q13.1 deletion was of maternal origin.

CONCLUSIONSNP array can accurately define the size, location and parental origin of chromosomal microdeletions, which may facilitate the diagnosis of AS due to 15q11q13 deletion and better understanding of its genotype-phenotype correlation.

Angelman Syndrome ; genetics ; Child ; Genotype ; Humans ; Karyotyping ; methods ; Male ; Phenotype ; Polymorphism, Single Nucleotide ; genetics

A reliable UPLC-MS method was developed for the simultaneous determination of 4 chemicals (Sudan Ⅰ, tetrabromobisphenol A, tris ( 1, 3-dichloroisopropyl ) phosphate and tris ( chlorisopropyl ) Phosphate) in children products. The samples were ultrasonic extracted with acetonitrile, and then the four chemicals were separated on a C18 column in 3 min. Results showed that the limit of quantification of the method was between 5 and 500μg/kg. The calibration curves were linear within 2-3 orders of magnitude with typical correlation coefficient above 0 . 9995 . The recoveries ranged from 83 . 7% to 97 . 8% with three addition levels. The sensitivity, recovery and selectivity of the method could fully meet the requirements of practical work.