1.Application of LIIR Automatic Analysis System of Infrared Spectroscopy in the determination of urinary stone composition
Xizhao SUN ; Luming SHEN ; Xiaoming CONG ; Yun WANG ; Jianlin Lü ; Lei HE
Chinese Journal of Urology 2011;32(1):24-26
Objective To determine the value of applying LIIR Automatic Analysis System of Infrared Spectroscopy in analyzing urinary stone composition. Methods 1450 samples of urinary stones were collected from 1032 male and 418 female patients. The age of patients ranged from 6 months to 88 years. The mean ages were 41.7±15.3 and 42.0±15.6 years for male and female patients, respectively. Of 1450 stones, 875 cases were located in kidney (60.34%), 504 cases in ureter (34.76%) and 71 cases in bladder (4.90%). All stones were analyzed by LIIR Automatic Analysis System of Infrared Spectroscopy (Tianjin). Analysis results were reevaluated by the artificial analysis of spectrogram, if necessary, with polarization microscope, chemical analysis, and X-ray diffraction.Results Calcium oxalate monohydrate stones were found in 714 cases (49. 24%), carbonate apatite stones in 444 cases (30.62%), anhydrous uric acid stones in 93 cases (6.41%), calcium oxalate dihydrate stones in 92 cases (6. 34 % ), ammonium magnesium phosphate hexahydrate stones in 28 cases (1.93%), cystine stones in 23 cases (1.59%), ammonium urate stones in 20 cases (1.38%), uric acid dihydrate stones in 16 cases (1.10%), brushite stones in 12 cases (0.83%), sodium urate monohydrate stones in 2 cases (0. 14%), calcium carbonate stones in 1 cases (0. 07%), and other stone types in 5 cases (0. 34%). Most urinary stones were composed of 2 or more compositions, and pure stones were only observed in 397 cases (27.38%). Most of the mixed stones contained calcium and non-calcium mixed stone was rarely observed. In addition, 15 stones were found in infants who had consumed melamine-contaminated milk powder. These stones were composed of uric acid dihydrate and ammonium urate. The results of reevaluation by artificial analysis showed the following: among pure and mixed stones, false detection occurred in 6 cases (0.41%), of which the composition was ammonium urate or carbonate apatite determined by automatic system but the true composition was anhydrous uric acid. False negative detection occurred in 9 cases (0.62%), of which the composition was ammonium magnesium phosphate hexahydrate or carbonate apatite in 7 cases, but in other 2 cases the composition could not be determined by artificial analysis. The false negative detection of components with relatively low content occurred in 6 cases and 10 cases in stones with 2 components and 3 components, respectively. The undetected composition in these cases was ammonium magnesium phosphate hexahydrate or carbonate apatite. Conclusion Automatic Analysis System of Infrared Spectroscopy has many advantages in accuracy, automation and is quick in analyzing the composition of urinary stones, and is worthy of promotion in clinical use.
2.A multicenter,double-blind, placebo-controlled, randomized, phase Ⅲ clinical study of etanercept in treatment of ankylosing spondylitis
Feng HUANG ; Jie ZHANG ; Jianlin HUANG ; Donghai WU ; Zhanguo LI ; Shunle CHEN ; Yunfeng PAN ; Li MA ; Shi CHEN ; Liangjing Lü ; Zunming YANG
Chinese Journal of Internal Medicine 2010;49(9):741-745
Objective To evaluate the short-term efficacy and safety of etanercept treatment in Chinese patients with active ankylosing spondylitis ( AS ). Methods This was a 12-week multicenter,double-blind, placebo-controlled, randomized phase Ⅲ clinical study. The first part was a 6-week placebocontrolled period followed by a 6-week open-label period. The primary efficacy endpoint was the percentage of subjects achieving a 20% improvement in assessment in ankylosing spondylitis (ASAS) ( ASAS 20). The secondary efficacy endpoints were the percentage of patients achieving a 40% improvement in ASAS (ASAS 40), achieving a 50% improvement in ASAS( ASAS 50), achieving a 70% improvement in ASAS (ASAS 70), and ASAS 5/6 responses at all visits, and the improvement in subject global assessment,physician global assessment, nocturnal and total back pain, bath AS functional index ( BASFI ), bath AS disease activity index (BASDAI), spinal mobility, joint assessment and quality of life assessment. All subjects in the study were evaluated for safety. Results The primary endpoint, ASAS 20 at week 6, was achieved by 86. 5% (64/74) patients in the etanercept group compared to 29. 5% (23/78) patients in the placebo group(P <0. 001 ). As early as week 2, the percentages of patients achieving the ASAS 20 between the two groups were significantly different. Furthermore, the majority of secondary efficacy end points were also significantly improved. Most of adverse events (AE) were mild in nature, the commonest adverse events were elevated liver function levels, injection site reactions and nasopharyngitis. No death or serious AE were observed. Conclusion Etanercept can improve symptoms fastly,significantly and safely in Chinese patients with active AS.
3.A recombinant adeno-associated virus expressing secretory TGF-β type Ⅱ receptor inhibits triple-negative murine breast cancer 4T1 cell proliferation and lung metastasis in mice
Zhi CUI ; Cuijiao MA ; Qianru WANG ; Jinhao CHEN ; Ziyang YAN ; Jianlin YANG ; Yafeng LÜ ; Chunyu CAO
Journal of Southern Medical University 2024;44(5):818-826
Objective To investigate the effects of an adeno-associated virus(AAV2)vector expressing secretory transforming growth factor-β(TGF-β)type Ⅱ receptor(sTβRⅡ)extracellular domain-IgG2a Fc fusion protein(sTβRⅡ-Fc)on proliferation and migration of triple-negative murine breast cancer 4T1 cells in mice.Methods The pAAV-sTβRⅡ-Fc vector expressing sTβRⅡ-Fc fusion protein constructed by molecular cloning,the capsid protein-expressing vector pAAV2 and the helper vector were co-transfected into HEK 293T cells to prepare the recombinant AAV2-sTβRⅡ virus,which was purified by density gradient centrifugation with iodixanol.Western blotting was used to examine the effects of AAV-sTβRⅡ virus on Smad2/3 phosphorylation in 4T1 cells and on expression levels of E-cadherin,vimentin and p-Smad2/3 in 4T1 cell xenografts in mice.BALB/c mice bearing subcutaneous xenografts of luciferase-expressing 4T1 cells received intravenous injections of AAV-sTβRⅡ virus,AAV-GFP virus or PBS(n=6)through the tail vein,and the proliferation and migration of 4T1 cells were analyzed with in vivo imaging.Ki67 expression in the tumor tissues and sTβRⅡ protein expressions in mouse livers were detected with immunohistochemistry and immunofluorescence staining,and tumor metastases in the vital organs were examined with HE staining.Results The recombinant pAAV-sTβRⅡ-Fc vector successfully expressed sTβRⅡ in HEK 293T cells.Infection with AAV2-sTβRⅡ virus significantly reduced TGF-β1-induced Smad2/3 phosphorylation in 4T1 cells and effectively inhibited proliferation and lung metastasis of 4T1 xenografts in mice(P<0.05).In the tumor-bearing mice,intravenous injection of AAV-sTβRⅡ virus significantly increased E-cadherin expression,reduced vimentin and Ki67 protein expressions and Smad2/3 phosphorylation level in the tumor tissues(P<0.05 or 0.01),and induced liver-specific sTβRⅡ expression without causing body weight loss or heart,liver,spleen or kidney pathologies.Conclusion The recombinant AVV2 vector encoding sTβRⅡ extracellular domain is capable of blocking the TGF-β signaling pathway to inhibit the proliferation and lung metastasis of 4T1 cells in mice.
4.A recombinant adeno-associated virus expressing secretory TGF-β type Ⅱ receptor inhibits triple-negative murine breast cancer 4T1 cell proliferation and lung metastasis in mice
Zhi CUI ; Cuijiao MA ; Qianru WANG ; Jinhao CHEN ; Ziyang YAN ; Jianlin YANG ; Yafeng LÜ ; Chunyu CAO
Journal of Southern Medical University 2024;44(5):818-826
Objective To investigate the effects of an adeno-associated virus(AAV2)vector expressing secretory transforming growth factor-β(TGF-β)type Ⅱ receptor(sTβRⅡ)extracellular domain-IgG2a Fc fusion protein(sTβRⅡ-Fc)on proliferation and migration of triple-negative murine breast cancer 4T1 cells in mice.Methods The pAAV-sTβRⅡ-Fc vector expressing sTβRⅡ-Fc fusion protein constructed by molecular cloning,the capsid protein-expressing vector pAAV2 and the helper vector were co-transfected into HEK 293T cells to prepare the recombinant AAV2-sTβRⅡ virus,which was purified by density gradient centrifugation with iodixanol.Western blotting was used to examine the effects of AAV-sTβRⅡ virus on Smad2/3 phosphorylation in 4T1 cells and on expression levels of E-cadherin,vimentin and p-Smad2/3 in 4T1 cell xenografts in mice.BALB/c mice bearing subcutaneous xenografts of luciferase-expressing 4T1 cells received intravenous injections of AAV-sTβRⅡ virus,AAV-GFP virus or PBS(n=6)through the tail vein,and the proliferation and migration of 4T1 cells were analyzed with in vivo imaging.Ki67 expression in the tumor tissues and sTβRⅡ protein expressions in mouse livers were detected with immunohistochemistry and immunofluorescence staining,and tumor metastases in the vital organs were examined with HE staining.Results The recombinant pAAV-sTβRⅡ-Fc vector successfully expressed sTβRⅡ in HEK 293T cells.Infection with AAV2-sTβRⅡ virus significantly reduced TGF-β1-induced Smad2/3 phosphorylation in 4T1 cells and effectively inhibited proliferation and lung metastasis of 4T1 xenografts in mice(P<0.05).In the tumor-bearing mice,intravenous injection of AAV-sTβRⅡ virus significantly increased E-cadherin expression,reduced vimentin and Ki67 protein expressions and Smad2/3 phosphorylation level in the tumor tissues(P<0.05 or 0.01),and induced liver-specific sTβRⅡ expression without causing body weight loss or heart,liver,spleen or kidney pathologies.Conclusion The recombinant AVV2 vector encoding sTβRⅡ extracellular domain is capable of blocking the TGF-β signaling pathway to inhibit the proliferation and lung metastasis of 4T1 cells in mice.