Urine proteomics identifies biomarkers for diabetic kidney disease at different stages.

BACKGROUND: Type 2 diabetic kidney disease is the most common cause of chronic kidney diseases (CKD) and end-stage renal diseases (ESRD). Although kidney biopsy is considered as the 'gold standard' for diabetic kidney disease (DKD) diagnosis, it is an invasive procedure, and the diagnosis can be influenced by sampling bias and personal judgement. It is desirable to establish a non-invasive procedure that can complement kidney biopsy in diagnosis and tracking the DKD progress. METHODS: In this cross-sectional study, we collected 252 urine samples, including 134 uncomplicated diabetes, 65 DKD, 40 CKD without diabetes and 13 follow-up diabetic samples, and analyzed the urine proteomes with liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS). We built logistic regression models to distinguish uncomplicated diabetes, DKD and other CKDs. RESULTS: We quantified 559 ± 202 gene products (GPs) (Mean ± SD) on a single sample and 2946 GPs in total. Based on logistic regression models, DKD patients could be differentiated from the uncomplicated diabetic patients with 2 urinary proteins (AUC = 0.928), and the stage 3 (DKD3) and stage 4 (DKD4) DKD patients with 3 urinary proteins (AUC = 0.949). These results were validated in an independent dataset. Finally, a 4-protein classifier identified putative pre-DKD3 patients, who showed DKD3 proteomic features but were not diagnosed by clinical standards. Follow-up studies on 11 patients indicated that 2 putative pre-DKD patients have progressed to DKD3. CONCLUSIONS: Our study demonstrated the potential for urinary proteomics as a noninvasive method for DKD diagnosis and identifying high-risk patients for progression monitoring.

HT-RCM: Hashimoto's Thyroiditis Ultrasound Image Classification Model Based on Res-FCT and Res-CAM.

The early lesions of Hashimoto's thyroiditis are inconspicuous, and the ultrasonic features of these early lesions are indistinguishable from other thyroid diseases. This paper proposes a Hashimoto Thyroiditis ultrasound image classification model HT-RCM which consists of a Residual Full Convolution Transformer (Res-FCT) model and a Residual Channel Attention Module (Res-CAM). To collect the low-order information caused by hypoechoic signals accurately, the residual connection is injected between FCTs to form Res-FCT which helps HT-RCM superimpose the low-order input information and high-order output information together. Res-FCT can make HT-RCM focus more on hypoechoic information while avoiding gradient dispersion. The initial feature map is inserted into Res-FCT again through a down-sampling component, which further helps HT-RCM exact multi-level original semantic information in the ultrasound image. Res-CAM is constructed by implementing a residual connection between a channel attention module and a convolution layer. Res-CAM can effectively increase the weights of the lesion channels while suppressing the weights of the noise channels, which makes HT-RCM focus more on the lesion regions. The experimental results on our collected dataset show that HT-RCM outperforms the mainstream models and obtains state-of-the-art performance in HT ultrasound image classification.

FBENet: Feature-Level Boosting Ensemble Network for Hashimoto's Thyroiditis Ultrasound Image Classification.

Distinguishing Hashimoto's thyroiditis (HT) lesions from ordinary thyroid tissues is difficult with ultrasound images. Challenges in achieving high performance of HT ultrasound image classification include the low resolution, blurred features and large area of irrelevant noise. To address these problems, we propose a Feature-level Boosting Ensemble Network (FBENet) for HT ultrasound image classification. Specifically, to capture the features of suspicious HT lesions efficiently, an Ensemble Feature Boosting Module (EFBM) is introduced into the feature-level ensemble to boost the blurred features. Then, the spatial attention mechanism is adopted in backbone models to improve the feature focusing performance and representation ability. Furthermore, feature-level ensemble technique is employed in the training process to achieve more comprehensive feature representation ability. Experimentally, FBENet was trained on 6,503 HT ultrasound images, and tested on 1,626 HT ultrasound images with 82.92% accuracy and 89.24% AUC on average.

Integrated metabolomics and transcriptomics analysis reveals the mechanism of Tangbi capsule for diabetic lower extremities arterial disease.

Objective: Tangbi capsule (TBC) is a traditional Chinese medicine prescription, which has the potential to improve the vascular insufficiency of lower extremities and limb numbness in diabetes. However, the potential mechanism remains unknown. This study aims to investigate the pharmacological effects and mechanism of TBC on rats with diabetic lower extremities arterial disease (LEAD). Methods: The mechanism of TBC on diabetic LEAD was investigated through metabolomics and transcriptomics analysis, and the main components of TBC were determined by mass spectrometry. The efficacy and mechanism of TBC on diabetic LEAD rats were investigated through in vitro experiments, histopathology, blood flow monitoring, western blot, and real-time polymerase chain reaction. Results: Mass spectrometry analysis identified 31 active chemical components in TBC including (2R)-2,3-Dihydroxypropanoic acid, catechin, citric acid, miquelianin, carminic acid, salicylic acid, formononetin, etc. In vitro analysis showed that TBC could reduce endothelial cell apoptosis and promote angiogenesis. Histopathological analysis showed that TBC led to an obvious improvement in diabetic LEAD as it improved fibrous tissue proliferation and reduced arterial wall thickening. In addition, TBC could significantly increase the expression levels of HIF-1α, eNOS, and VEGFA proteins and genes while reducing that of calpain-1 and TGF-ß, suggesting that TBC can repair vascular injury. Compared with the model group, there were 47 differentially expressed genes in the whole blood of TBC groups, with 25 genes upregulated and 22 downregulated. Eighty-seven altered metabolites were identified from the serum samples. Combining the changes in differentially expressed genes and metabolites, we found that TBC could regulate arginine biosynthesis, phenylalanine metabolism, pyrimidine metabolism, arachidonic acid metabolism, pyrimidine metabolism, arachidonic acid metabolism, nucleotide metabolism, vitamin B6 metabolism and other metabolic pathways related to angiogenesis, immune-inflammatory response, and cell growth to improve diabetic LEAD. Conclusion: TBC improved vascular endothelial injury, apoptosis, lipid accumulation, liver and kidney function, and restored blood flow in the lower extremities of diabetic LEAD rats. The mechanism of TBC in the treatment of diabetic LEAD may be related to the modulation of inflammatory immunity, lipid metabolism, and amino acid metabolism. This study presented preliminary evidence to guide the use of TBC as a therapy option for diabetic LEAD.

Effect and Safety of Herbal Medicine Foot Baths in Patients with Diabetic Peripheral Neuropathy: A Multicenter Double-Blind Randomized Controlled Trial.

OBJECTIVE: To evaluate the effect and safety of foot baths with Tangbi Waixi Decoction (TW) in treating patients with diabetic peripheral neuropathy (DPN). METHODS: It is a multicenter double-blinded randomized controlled trial. Participants with DPN were recruited between November 18, 2016 and May 30, 2018 from 8 hospitals in China. All patients received basic treatments for glycemic management. Patients received foot baths with TW herbal granules either 66.9 g (intervention group) or 6.69 g (control group) for 30 min once a day for 2 weeks and followed by a 2-week rest, as a therapeutic course. If the Toronto Clinical Scoring System total score (TCSS-TS) ⩾6 points, the patients received a total of 3 therapeutic courses (for 12 weeks) and were followed up for 12 weeks. The primary outcome was change in TCSS-TS score at 12 and 24 weeks. Secondary outcomes included changes in bilateral motor nerve conduction velocity (MNCV) and sensory nerve conduction velocity (SNCV) of the median and common peroneal nerve. Safety was also assessed. RESULTS: Totally 632 patients were enrolled, and 317 and 315 were randomized to the intervention and control groups, respectively. After the 12-week intervention, patients in both groups showed significant declines in TCSSTS scores, and significant increases in MNCV and SNCV of the median and common peroneal nerves compared with pre-treatment (P<0.05). The reduction of TCSS-TS score at 12 weeks and the increase of SNCV of median nerve at 24 weeks in the control group were greater than those in the intervention group (P<0.05). The number of adverse events did not differ significantly between groups (P>0.05), and no serious adverse event was related with treatment. CONCLUSION: Treatment of TW foot baths was safe and significantly benefitted patients with DPN. A low dose of TW appeared to be more effective than a high dose. (Registry No. ChiCTR-IOR-16009331).

Molecular complexity analysis of the diagnosis of Gitelman syndrome in China.

Gitelman syndrome (GS) is an autosomal recessive renal tubal disease characterized by hypomagnesemia, hypokalemia, and hypocalciuria. The disease is caused by defects in the SLC12A3 gene, which encodes the thiazide diuretic-sensitive sodium chloride cotransporter (NCCT). In this study, a 20-year-old female patient with recurrent hypokalemia was tested for a hypokalemia-related panel using Next Generation Sequencing. Pedigree analysis was performed on her parents (non-consanguineous) and sister using Sanger sequencing. The results revealed that the patient carried compound heterozygous variants of the SLC12A3 gene: c.179C > T (p.T60M) and c.1001G > A (p.R334Q). Furthermore, her asymptomatic 6-year-old sister also carried both mutations. While the p.T60M mutation had been reported previously, the p.R334Q mutation was novel, and amino acid position 334 was identified as a mutation hotspot. Our findings provide an accurate molecular diagnosis that is essential for the diagnosis, counseling, and management of not only the symptomatic patient but also her asymptomatic sister. This study contributes to our understanding of the GS, which has a prevalence of approximately 1 in 40,000 and a heterozygous mutation carrier rate of 1% in Caucasians. Specifically, we observed a compound heterozygous mutation of the SLC12A3 gene in a 20-year-old female patient presenting with clinical symptoms consistent with GS.

Short-Term Effect of Continuous Subcutaneous Insulin Infusion and Multiple Daily Injection in Perioperative Patients with Type 2 Diabetes Mellitus.

Background: Hyperglycemia is common and difficult to control in perioperative patients with type 2 diabetes mellitus (T2DM), which impacts their prognosis after operation. Our study investigated the short-term effect of continuous subcutaneous insulin infusion (CSII) and multiple daily injection (MDI) in perioperative T2DM patients using the data envelopment analysis (DEA). Methods: T2DM patients (n = 639) who underwent surgeries in Guangdong Provincial Hospital of Traditional Chinese Medicine (2009.01-2017.12) were included. Insulin was provided to each patient during the study and separated into a CSII group (n = 369) and an MDI group (n = 270). DEA was performed to compare the therapeutic indexes and investigate the short-term effect of the CSII group and MDI group. Results: Scale efficiencies of the CSII group with CCR model and BCC model were better than that of the MDI group. Regarding slack variables, with higher surgical levels, the CSII group was closer to the ideal state than the MDI group, which indicated in improving the average fasting blood glucose (AFBG), antibiotic use days (AUD), preoperative blood glucose control time (PBGCT), first postoperative day fasting blood glucose (FPDFBG), and postoperative hospitalization days (PHD). Conclusion: CSII could effectively control blood glucose levels and shorten perioperative hospitalizing time for T2DM patients, indicating that CSII was beneficial in perioperative period and should be promoted clinically.

HTC-Net: Hashimoto's thyroiditis ultrasound image classification model based on residual network reinforced by channel attention mechanism.

Convolutional neural network (CNN) is efficient in extracting and aggregating local features in the spatial dimension of the images. However, obtaining the inapparent texture information of the low-echo area in the ultrasound images is not easy, and it is especially challenging for the early lesion recognition in Hashimoto's thyroiditis (HT) ultrasound images. In this paper, a HT ultrasound image classification model HTC-Net based on residual network reinforced by channel attention mechanism is proposed. HTC-Net strengthens the features of the important channels by reinforced channel attention mechanism through which the high-level semantic information is enchanced and the low-level semantic information is suppressed. Residual network assists HTC-Net focus on the key local areas of the ultrasound images while pay attention to the global semantic information. Furthermore, in order to solve the problem of uneven distribution caused by large amount of difficult-to-classify samples in the data sets, a new feature loss function TanCELoss with weight factor dynamically adjusting is constructed. TanCELoss function can better assist HTC-Net to transform difficult-to-classify samples into easy-to-classify samples gradually, and improve the balancing distribution of the samples. The experiments are implemented based on data sets collected by the Endocrinology Department of four branches from Guangdong Provincial Hospital of Chinese Medicine. Both quantitative testing and visualization results show that HTC-Net obtains STOA performance for early lesions recognition in HT ultrasound images. HTC-Net has great application value especially under the condition of owning only small data samples.

[Comparative study of enhancing effect on mRNA expression of hematopoietic growth factors in rat bone marrow mesenchymal stem cells by ginseng polysaccharide and ginsenoside].

OBJECTIVE: To study mRNA expression levels of main hematopoietic growth factors in bone marrow mesenchymal stem cells (BM-MSC), and to compare effect on mRNA expression levels treated by ginseng polysaccharide and ginsenoside. METHODS: Relative quantification real-time polymerase chain reaction (RT-PCR) was used to observe mRNA expression levels of IL4, Csf2, Kitlg, Csf1, IL6, Lif, Csf3, IL11, Epo, and IL3, etc. in rat BM-MSC treated with ginseng polysaccharide (20 microg/mL) or ginsenoside (20 microg/mL) at 12, 24, and 36 h. RESULTS: IL4 and Csf2 mRNA expressions were not detected. Relative expression of Kitlg, Csf1, IL6, Lif, Csf3, IL11, Epo and IL3 mRNA ranked in an attenuating order when compared with Gapdh mRNA. mRNA expression of Epo and IL3 was not significantly changed at any time point by treatment of ginseng polysaccharide or ginsenoside in rat BM-MSC (P > 0.05). mRNA expression of Csf1, IL6, Lif, Csf3 and IL11 were significantly enhanced at 12 and 36 h by treatment of ginseng polysaccharide (P < 0.05) and that of Csf1, IL6, Lif, Csf3, and Kitlg were significantly enhanced at 24 h in rat BM-MSC (P < 0.05). The enhanced mRNA expression was Csf3 at 12 h, Csf3, IL6 and Lif at 24 h, and Csf3, IL6, Lif, IL11, and Kitlg, respectively at 36 h by treatment of ginsenoside in rat BM-MSC. CONCLUSIONS: The enhancement of ginseng polysaccharide was stronger than that of ginsenoside on mRNA expression of hematopoietic growth factors in the initial stage. As time went by, the enhancement of ginsenoside gradually increased and exceeded that of ginseng polysaccharide.