Objective To determine the changes in serum and urine vitamin D binding protein ( VDBP) concentrations in type 2 diabetes, and to explore the clinical significance. Methods The serum and urine VDBP concentrations in 102 healthy individuals and 106 type 2 diabetic patients were determined by ELISA. For analysis and comparison, 106 type 2 diabetic patients were divided into imperfect glycemic control subgroup and perfect glycemic control subgroup, microalbuminuria subgroup and normal albuminuria subgroup. Results The cut-off point of serum VDBP concentrations was 60. 6 μg/ ml and the cut-off point of the urine ratio of VDBP and creatinine was 7. 76 mg/ g, and both were determined according to the upper limit of 97. 5 % credit intervals in 110 healthy individuals. Serum VDBP concentration and the urine ratio of VDBP to creatinine in type 2 diabetic patients were significantly higher than those in the healthy individuals ( P < 0. 01 ), the imperfect glycemic control subgroup had higher serum VDBP concentrations and the urine ratio of VDBP to creatinine than those in the perfect glycemic control subgroup ( P <0. 05). The microalbuminuria subgroup had higher urine ratio of VDBP to creatinine than that in the normal albuminuria subgroup ( P<0. 01). Urine ratio of VDBP to creatinine in diagnosing early diabetic nephropathy had sensitivity of 96. 4 % , specificity of 68 % , and concordance of 83% . Conclusion Detection of serum VDBP levels has some reference value in understanding the state of diabetes. Combined determinations of urine ratio of VDBP to creatinine and ratio of albumin to creatinine have significant clinical value in the early diagnosis of diabetic nephropathy.

Objective: To investigate the role of piperine on the transformation of endothelial cells into fibroblasts. Methods: Cultured human umbilical vein endothelial cells (HUVECs, 4-6 passage) were used for the main experiments. The transformation models of endothelial cells into fibroblasts were induced by transforming growth factor β (TGF-β) stimulation. HUVECs were divided into 6 groups: control group, TGF-β group and 4 groups treated with various concentrations of piperine (1, 5, 10, 20 μmol/L). CKK-8 was used to detect cell proliferation. The CD31/α-smooth muscle actin (α-SMA) expression level was detected by fluorescent staining. The vascular endothelial cadherin (VE-cadherin)/vimentin expression was detected by immunofluorescence staining. RT-PCR was used detect the mRNA expressions of transformation markers. Western blot was used to detect the protein expression of snail and twist. Results: TGF-β increased HUVECs proliferation (P<0.05), which could be significantly inhibited by 10 and 20 μmol/L of piperine, but not by 1 and 5 μmol/L of piperine. Immunofluorescence results demonstrated that TGF-β increased HUVECs transformation to fibroblasts as shown by downregulated expression of endothelial markers CD31, VE-cadherin, and upregulated expression of α-SMA and vimentin, again, these effects could be attenuated by 10 and 20 μmol/L piperine. The expression levels of collagen type Ⅰ and type Ⅲ were significantly higher in TGF-β group than in control group (P<0.05), significantly lower in TGF-β+10 μmol/L piperine group and TGF-β+20 μmol/L piperine group than in TGF-β group (P<0.05).In addition, RT-PCR results showed that TGF-β increased mRNA expression of transformation markers (snail1, snail2, twist1, twist2), while 10 and 20 μmol/L of piperine could significantly downregulated the mRNA expressions of these markers. The protein expression levels of snail and twist were significantly higher in TGF-β group than in control group (both P<0.05), which was significantly lower in TGF-β+20 μmol/L piperine group than in TGF-β group (both P<0.05). Conclusions Piperine can inhibit the transformation of endothelial cells into fibroblasts. This effect might be viewed as one of the potential mechanisms of reduced myocardial fibrosis post piperine treatment.

Objective To investigate the polymorphisms of human cytomegalovirus ( HCMV ) UL146 gene in asymptomatic children. Methods Urine samples were collected from 47 asymptomatic chil-dren who were positive for HCMV DNA. PCR was performed to amplify the open reading frame ( ORF) of UL146 gene. Positive bands were sequenced and variations in UL146 gene were analyzed by using bioinfor-matics software. Results Seventeen samples were successfully amplified and sequenced. Variations spread all over the sequence of UL146 gene and the variability in nucleotide and amino acid sequences ranged from 0％ to 42. 5％ and 0％ to 67. 7％ respectively. Compared with the Towne strain, there was diversity in sig-nal sequence and C-terminal region. Phylogenetic analysis indicated that UL146 in the 17 asymptomatic chil-dren belonged to four genotypes, which were G1, G8, G9 and G11. Forms of post-translational modification varied greatly among the four genotypes, while the important functional region of ELRCXC chemokine was highly conservative. Secondary structure prediction showed that random-coli conformation was the predomi-nant structure of active proteins. Isoelectric point ( PI) and molecular weight ( MW) were dissimilar among the four genotypes. Conclusion HCMV UL146 gene in asymptomatic children was hypervariable in both nucleotide sequence and amino acid structure. However, the important functional region was highly con-served. The predominant genotypes of UL146 in these children were G1, G8, G9 and G11, and the geno-type distribution in them showed no significant difference with previous findings in children with symptomatic HCMV infection.

Objective:To observe the presence or absence of necroptosis in PC12 cells after radiation injury, and to detect the expression of receptor-interacting protein 3(RIP3) and evaluate its regulatory effect on necroptosis.Methods:PC12 cells were treated with different doses of irradiation and their necroptosis was detected by lactate dehydrogenase (LDH) release at different time points. After pretreatment with necroptosis inhibitor Necrostatin-1(Nec-1), the changes of cell necroptosis were detected by LDH. The expression level of RIP3 after irradiation intervention was detected by Western blot (WB). After pretreatment with the RIP3-specific inhibitor GSK′872, the changes of cell necroptosis were detected by LDH. The best transfection sequence of RIP3 knockout was screened by WB. The cells were divided into the control group, irradiation group, solvent control group, no-load control group and pretreatment group. WB, immunofluorescence staining, MTT, LDH and Annex V-fluorescein Isothiocyanate/Propidium Iodide (AnnexV-FITC/PI) flow cytometry were used for detection and analysis.Results:After 4 Gy irradiation, the degree of cell necrosis was the highest after 3 hours of culture, and the expression level of RIP3 protein was up-regulated. The cell necrosis was decreased after Nec-1, GSK′872 and RIP3 gene knockdown pretreatment.Conclusions:The radiation injury of 4 Gy can induce the necroptosis of PC12 cells, and the most significant effect can be observed when cultured for 3 hours after irradiation. RIP3 is involved in the process of necroptosis of PC12 cells induced by radiation injury, and plays a pivotal positive regulatory role.

Objective: By detecting the expression of interleukin-13 (IL-13) and periostin in the airway of asthmatic patients, the pathological changes and pulmonary functions of airway tissues in asthmatic patients were evaluated, and the role of IL-13 and periostin airway remodeling in bronchial asthma was preliminarily explored. Methods: The bronchial tissues adjacent to tumor nest were obtained from 12 patients with lung cancer complicated with bronchial asthma (asthmatic group) and 12 lung cancer patients without bronchial asthma (non-asthmatic group) after lung cancer resection. Pulmonary function was measured for all subjects before surgery. Pathological changes of airway tissues and degree of airway remodeling were assessed by hematoxylin-eosin (H&E) staining, masson′s trichrome staining, and periodic acid-silver methenamine (PASM) staining of paraffin-embedded sections. The expression of IL-13 and periostin in bronchial tissues were evaluated by immunohistochemistry. Results: Values of the forced expiratory volume in 1 second of the predicted value (FEV1% pred) and FEV1/forced vital capacity (FEV1/FVC%) in asthmatic patients were significantly decreased compared with the non-asthmatic patients (P<0.05), indicating that lung function was impaired in asthmatic patients. There was more severe airway remodeling representing as thickening of basement membranes, collagen deposition, and increasing of goblet cells and fibroblasts in asthmatic patients than in non-asthmatic patients (all P<0.05). The expression of IL-13 and periostin were higher in asthmatic tissues than in non-asthmatic tissues (P<0.05). The immunohistochemical expression of IL-13 and periostin in bronchial tissues were positively correlated with the degree of airway remodeling in asthmatic patients, and the expression of IL-13 and periostin in bronchial tissues were positively correlated with each other. Conclusions The expression of IL-13 and periostin were increased in bronchial tissue in patients with asthma. They work together to promote the occurrence of airway remodeling, which eventually lead to a decline in lung function.