Objective To investigate the correlation between the level of cystatin C (Cys C) and different body mass index (BMI) in patients with type 2 diabetes mellitus (T2DM).Methods The clinical data of 160 patients with T2DM was retrospectively analyzed.They were divided into normal weight (NW) group (n =88),over weight (OW) group (n =42) and obesity (OB) group (n =30).NW and OB were defined as BMI ＜ 25 kg/m2 and BMI ≥ 28 kg/m2 according to the WHO-Western Pacific Region diagnostic set forth in 2000.The basic clinical data and biochemical indexes of all the subjects were collected.Spearman correlation analysis and Logistic regression analysis were used to evaluate the influencing factors for the different BMI in T2DM.Results Compared with T2DM-NW group,the level of serum Cys C was significantly increased in T2DM-OW group and T2DM-OB group,and Cys C in T2DM-OB group was significantly higher than T2DM-OW group (P ＜0.05 or P ＜ 0.01).T2DM-NW/OW/OB was positively correlated with BMI,hypertension,ALT,AST,TC,TG,LDL-C,ApoB and Cys C (P ＜ 0.05 or P ＜ 0.01).Logistic regression analysis showed that ALT,TG,LDL-C and Cys C were the independent risk factors for T2DM with OW/OB (P＜ 0.05 or P ＜ 0.01).Conclusion With increasing of BMI in T2DM,serum Cys C level gradually increase.It suggests that the increased Cys C maybe play an important role in the pathogenesis of the different BMI in T2DM.

Objective:To analyze the clinical features and genetic background of adrenoleukodystrophy(ALD).Methods:In this study, we reported a rare case of ALD who initially presented with progressive bilateral lower limb weakness. The clinical data of the patient and his family members were collected and the ABCD1 gene was sequenced for the patient and his three daughters by a high-throughput sequencing method.Results:The proband had a later onset of symptoms, a prolonged course of the disease, and initially exhibited bilateral lower limb weakness and nocturnal muscle spasms. The disease progressed to spastic quadriplegia, aphasia, dementia, swallowing difficulties, and urinary and fecal incontinence. Serum very-long-chain fatty acid concentrations were elevated. Subclinical cortisol secretion abnormalities were observed. Cranial imaging indicated symmetrical reduction in density around the lateral ventricles and white matter degeneration. The proband′s ABCD1 gene analysis revealed a novel heterozygous mutation c. 1367G>A, p. R456H. His three daughters carried the same nucleotide heterozygous mutation.Conclusion:This study investigates the clinical characteristics of ALD, providing additional clinical evidence for the diagnosis and treatment of this condition. Additionally, a novel mutation in the ABCD1 gene was identified, contributing to the genetic variation database.

Objective To investigate the correlation between the level of cystatin C (Cys C) and different body mass index (BMI) in patients with type 2 diabetes mellitus (T2DM).Methods The clinical data of 160 patients with T2DM was retrospectively analyzed.They were divided into normal weight (NW) group (n =88),over weight (OW) group (n =42) and obesity (OB) group (n =30).NW and OB were defined as BMI ＜ 25 kg/m2 and BMI ≥ 28 kg/m2 according to the WHO-Western Pacific Region diagnostic set forth in 2000.The basic clinical data and biochemical indexes of all the subjects were collected.Spearman correlation analysis and Logistic regression analysis were used to evaluate the influencing factors for the different BMI in T2DM.Results Compared with T2DM-NW group,the level of serum Cys C was significantly increased in T2DM-OW group and T2DM-OB group,and Cys C in T2DM-OB group was significantly higher than T2DM-OW group (P ＜0.05 or P ＜ 0.01).T2DM-NW/OW/OB was positively correlated with BMI,hypertension,ALT,AST,TC,TG,LDL-C,ApoB and Cys C (P ＜ 0.05 or P ＜ 0.01).Logistic regression analysis showed that ALT,TG,LDL-C and Cys C were the independent risk factors for T2DM with OW/OB (P＜ 0.05 or P ＜ 0.01).Conclusion With increasing of BMI in T2DM,serum Cys C level gradually increase.It suggests that the increased Cys C maybe play an important role in the pathogenesis of the different BMI in T2DM.

OBJECTIVETo investigate 4 populations of 80 samples of Vitex trifolia var. simplicifolia collected from Shandong and Jiangxi province and analyze their intraspecies genetic variance.

METHODInter-simple sequence repeat (ISSR) technique was applied for the study.

RESULTFifteen specific and stable primers were selected from 100 primers. A total of 129 sites were generated, and 115 of them (89.15%) were polymorphic. The data analyzed by PopGene demonstrated that the average polymorphic site percentage among the four populations was 71.89%. The average Shannon's information index was 0. 220 4. According to cluster analysis and the law of geographic variation, the populations were classified into two large groups: the Shandong group and the Jiangxi group.

CONCLUSIONThese results will provide the information for protection and utilization of V. trifolia var. simplicifolia and also further data for the study of genetic variation and species differentiation of V. trifolia var. simplicifolia.

Objective:To establish a rat model of neurogenic bladder and analyze the changes in kidney morphology and function and the expression of proteins in AngiotensinⅡ(AngⅡ)/transforming growth factor β1 (TGF-β1)/Smads pathway.Methods:Sprague-Dawley rats were randomly divided into experimental group (spinal nerve amputation, n=36) and control group (sham operation, n=12). At 6, 12, and 24 weeks, the bladder compliance was measured by cystometry, the kidney morphology was detected by B-ultrasound, blood urea nitrogen (BUN) and serum creatinine (Scr) in blood samples were examined, the kidney pathological changes were detected by Masson and HE staining, the distribution of AngⅡ/TGF-β1/Smads pathway proteins was analyzed by immunohistochemisty, and the protein expressions in kidney were detected by Western blotting. Results:Urodynamics showed that the basic bladder pressure in experimental group was higher than that in control group. B-ultrasound showed that compared with the control group, the diameter of the renal pelvis of the rats with nerve dissection gradually increased ( P<0.05), and the hydronephrosis was gradually obvious. Compared with the control group, the BUN and Scr in experimental group gradually increased (both P<0.01). Masson and HE staining showed that compared with the control group, the collagen expression and renal tubulointerstitial scores in experimental group were gradually increased (both P<0.01). Immunohistochemisty showed that compared with the control group, in experimental group the expression of angiotensinⅡ receptor type 1 (AT1), TGF-β receptor 1(TGF-βR1), phosphorylated Smad2 gradually increased (all P<0.01), the pathway inhibitor Smad6 gradually decreased ( P<0.01), and the distribution of each protein in kidney was consistent. Western blotting showed a corresponding expression trend with immunohistochemisty. Conclusions:In neurogenic bladder caused by bilateral spinal nerve amputation, due to bladder dysfunction, increased bladder pressure induces hydronephrosis, destruction of the nephron structure, activation of AngⅡ/TGF-β1/Smads pathway, and renal fibrosis. This method is effective and has clinical similarities, laying a foundation for exploring neurogenic bladder treatment.

Objective:To assess the value of urine heat-shock protein-70 (HSP-70) in the early diagnosis of acute kidney injury (AKI) after cardiac cardiopulmonary bypass (CPB).Methods:Patients with cardiopulmonary bypass from May 2018 to July 2018 in Henan Provincial People's Hospital were enrolled as subjects. Urine samples were collected before and after cardiopulmonary bypass at 0 h, 2 h, 4 h, 6 h, 8 h, 12 h, 24 h and 48 h. Patients were divided into AKI group and non-AKI group according to the Kidney Disease: Improving Global Outcomes Guide. Urinary HSP-70, tissue inhibitor of metalloproteinase-2 (TIMP-2) and insulin-like growth factor-binding protein 7 (IGFBP7) were detected by enzyme-linked immunosorbent assay (ELISA) and urine neutrophil gelatinase-associated lipocalin (NGAL) was determined by immunoturbidimetry. The receiver operating characteristic (ROC) curve was plotted to calculate the critical value, sensitivity and specificity of urine HSP-70, [TIMP-2]×[IGFBP7] and NGAL for the diagnosis of postoperative AKI after CPB.Results:A total of 45 patients were enrolled in the study. There were 24 cases in AKI group and 21 cases in non-AKI group. The level of urinary HSP-70, [TIMP-2]×[IGFBP7] and NGAL in AKI group were significantly higher than in the non-AKI group at each postoperative time point, with statistically significant differences (all P<0.05). The level of urinary HSP-70 in AKI group peaked at 2 h after CPB, which was significantly earlier than the peak time of urine [TIMP-2]×[IGFBP7] and urine NGAL (12 h after CBP and 4 h after CBP, respectively). Urinary HSP-70≥2.1 μg/L could predict postoperative AKI of CPB at 2 h after CPB, with the area under the curve ( AUC) of 1.00, the sensitivity of 100.0% and the specificity of 100.0%. Urinary [TIMP-2]×[IGFBP7]>19.1 μg 2/L 2 could predict postoperative AKI of CPB at 12 h after CPB with the AUC of 0.94, the sensitivity of 87.5%, and the specificity of 100.0%. Urinary NGAL>27.4 μg/L could predict postoperative AKI of CPB at 4 h after CPB with the AUC of 0.95, the sensitivity of 95.8%, and the specificity of 85.7%. The positive predictive value of urine HSP-70≥2.1 μg/L at 2 h after CPB was 100.0%, and the negative predictive value was 100.0%. Conclusions:The level of urinary HSP-70 increases earlier than that of urinary [TIMP-2]×[IGFBP7] and NGAL in patients with AKI after CPB. Clinical monitoring of urinary HSP-70 level contributes to early diagnosis of AKI.

ObjectiveTo analyze and determine the differential components of freeze-dried and sun-dried Panacis Quinquefolii Radix（PQR）， and to compare the differences in their pro-angiogenic activities. MethodFingerprints of freeze-dried and sun-dried PQR were established based on ultra performance liquid chromatography（UPLC）， and chemometrics methods such as principal component analysis（PCA） and orthogonal partial least squares-discriminant analysis（OPLS-DA） were combined to determine the differential saponin composition of the two decoction pieces， and six representative saponins were selected and their contents in freeze-dried and sun-dried PQR were determined by UPLC. Transgenic zebrafish line Tg（fli1a∶EGFP） embryos fertilized for 24 h were selected， and different doses of 70% methanol extracts of freeze-dried and sun-dried PQR（10， 30 mg·L^-1） were used to intervene in normal zebrafish and in a zebrafish model of intersegmental vascular（ISV） injury induced by vascular endothelial growth factor（VEGF） receptor tyrosine kinase inhibitor Ⅱ（PTK787）， then the development of subintestinal vein（SIV） and ISV of zebrafish was observed， SIV diameter， mean number of crossings and mean number of germinations were determined， and the ISV vascular index was calculated， in order to compare the pro-angiogenic activities of the two decoction pieces. ResultThe similarity of the fingerprints of freeze-dried and sun-dried PQR decoction pieces was>0.950， and 17 common peaks were identified， of which 6 common peaks were designated as peak 6（ginsenoside Rg₁）， peak 7（ginsenoside Re）， peak 8（ginsenoside Rb₁）， peak 11（ginsenoside Rc）， peak 13（ginsenoside Rb₂）， and peak 16（ginsenoside Rd）， respectively. A total of 11 differential saponin components were screened by PCA and OPLS-DA， indicating that there were some differences in the contents of the components in the two decoction pieces. The results of determination showed that the contents of ginsenosides Rg₁， Re， Rb₁ and Rb₂ in freeze-dried PQR were higher than those in sun-dried PQR， while the contents of ginsenosides Rc and Rd were lower than those in sun-dried PQR（P<0.05， P<0.01）. In the study of the pro-angiogenic effect on normal zebrafish embryos， compared with the blank group， and the SIV vessel diameter， mean germination rate and mean crossover rate were significantly higher in the high-dose groups of freeze-dried and sun-dried PQR（P<0.01）， and the vessel diameter， mean numbers of crossings and germinations in the freeze-dried PQR group were higher than those of the sun-dried PQR group（P<0.05）. In the study of the pro-angiogenic effect on zebrafish embryos with ISV injury， the development of ISV in the model group was significantly inhibited when compared with the blank group， compared with the model group， different dose groups of freeze-dried and sun-dried PQR could promote the growth and sprouting of ISV， and the number of normal blood vessels in the freeze-dried PQR group was significantly higher than that in the sun-dried PQR group at the same dosage（P<0.05）. ConclusionFreeze-drying can effectively avoid the loss and secondary transformation of ginsenosides in PQR， and its angiogenic activity is better than that of sun-dried PQR， which can provide a reference for the production and development of high-quality PQR decoction pieces.

ObjectiveTo analyze the quantity-quality transfer of standard decoction of Ginseng Radix et Rhizoma（GRR） decoction pieces produced by fresh and traditional cutting， and to provide reference for quality control and application development of the decoction pieces produced by fresh cutting. MethodTen batches of representative GRR decoction pieces produced by fresh and traditional cutting and their standard decoctions were prepared by standard process， and high performance liquid chromatography（HPLC） fingerprint of the standard decoction was established and performed on an Agilent EC-C₁₈ column（4.6 mm×150 mm， 2.7 μm） with acetonitrile（A）-0.1% phosphoric acid aqueous solution（B） as the mobile phase for gradient elution（0-23 min， 18%-21%A； 23-35 min， 21%-28%A； 35-80 min， 28%-32%A）， and the detection wavelength was 203 nm. Then similarity evaluation， principal component analysis（PCA） and partial least squares-discriminant analysis（PLS-DA） of fingerprint of the standard decoction were performed to screen the differential components with variable importance in the projection（VIP） value>1. Quantitative analysis was carried out on the screened known differential components， and combined with the indicators of the dry extract rate and the transfer rate， to explore the differences in the quantity-quality transfer between the standard decoction of GRR decoction pieces produced by fresh and traditional cutting. ResultThe fingerprint similarity of the standard decoction of GRR decoction pieces produced by fresh and traditional cutting was more than 0.950， and 18 common peaks were identified， including 9 identified common peaks. The results of PCA and PLS-DA showed that there were some differences in the contents of index components between the two standard decoctions. The contents of ginsenoside Rg₁， Re and Ro in GRR decoction pieces produced by fresh cutting were higher than those in traditional decoction pieces， while the contents of ginsenoside Rb₁， Rc ， Rb₂ and Rd were lower than those in traditional decoction pieces. The contents of ginsenoside Rg₁， Re， Rb₁ and Ro in the standard decoction of GRR decoction pieces produced by fresh cutting were higher than those in the standard decoction of traditional decoction pieces， while the contents of ginsenoside Rc ， Rb₂ and Rd were comparable between the two standard decoctions. Compared with the standard decoction of the traditional decoction pieces， the average transfer rates of ginsenoside Rg₁， Rb₁， Rc， Rb₂ and dry extract rate of the standard decoction of GRR decoction pieces produced by fresh cutting were significantly increased（P<0.05）， and the average transfer rate of ginsenoside Re and Rd also increased， but the difference was not statistically significant. ConclusionThe dry extract rate， content and transfer rate of index components of standard decoction of GRR decoction pieces produced by fresh cutting are better than those of the standard decoction of traditional decoction pieces， which can provides data support for the subsequent clinical application of fresh cutting products.

Cullin-RING ligases (CRLs) recognize and interact with substrates for ubiquitination and degradation, and can be targeted for disease treatment when the abnormal expression of substrates involves pathologic processes. Phosphorylation, either of substrates or receptors of CRLs, can alter their interaction. Phosphorylation-dependent ubiquitination and proteasome degradation influence various cellular processes and can contribute to the occurrence of various diseases, most often tumorigenesis. These processes have the potential to be used for tumor intervention through the regulation of the activities of related kinases, along with the regulation of the stability of specific oncoproteins and tumor suppressors. This review describes the mechanisms and biological functions of crosstalk between phosphorylation and ubiquitination, and most importantly its influence on tumorigenesis, to provide new directions and strategies for tumor therapy.