2.Development of visible protein microarrays based on GoldMag particles-labeling technique
Jinfan TANG ; Qiang JIAN ; Yali CUI ; Chao CHEN ; Zheng LI
Chinese Journal of Clinical Laboratory Science 1985;0(04):-
Objective To develop a visible detection system prepared by protein microarrays which was based on GoldMag particles-labeled technique and compare the results of detection using the protein which was labeled with GoldMag particles and colloidal gold.Methods Human IgG was printed on the glass slides modified with epoxy groups,and goat anti-human IgG conjugated with GoldMag particles and colloidal gold respectlively was then added on the slide.The glass slides were incubated,and then the black images of microarray spots were produced by immuno-gold-silver staining method and observed by naked eyes and recorded with common flatbed scanner.Results The high signal-to-noise ratio could be obtained when the optimized procedures of GoldMag particles-labeling probe were introduced to the protein chip.The conditions of optimum assay were as follows:the spotting concentration of human IgG was 0.2 mg/ml,the glass slides were incubated at 37 ℃ for 2 hour to immobilize human IgG,and the silver enhancement time was 10 min-15 min.Parallelly,the optimized conditions for colloidal gold were as follows:the spotting concentration of human IgG was 0.1 mg/ml,the slides were incubated at 37 ℃ for 1 hour to immobilize human IgG,and the silver enhancement time was 15 min-20 min.Conclusions GoldMag particles-labeled protein technique is comparable to colloidal gold in applying to protein microarrays.The method is considerably simple and practical,and the protein labeled by GoldMag particles could be quantitative.
3.Analysis of risk factors related to venous thromboembolism after knee arthroscopy
Chao LIN ; Tao LIU ; Qiang REN ; Jian LI
China Journal of Endoscopy 2017;23(5):64-68
Objective To determine the risk factors related to venous thromboembolism after knee arthroscopy. Methods A retrospective study including patients from Feb 2012 to Mar 2016 was carried out to analyze the risk factors of venous thromboembolism after knee arthroscopy. A 1 : 2 matched control group was generated according to the surgeon and the date. Preoperative and perioperative data were collected with respect to age, gender, body mass index, smoking, blood fat, surgical procedure, the time length of ligating tourniquet, anesthesia methods, surgery time and Caprini evaluation. Univariate and multivariate analyses were performed. Results 29 cases of venous thromboembolic events (VTEs) occurred, resulting in an incidence rate of 1.17%. Factors associated with an elevated risk of postoperative VTEs included age OR^ =1.09, 95%CI (1.03, 1.16) and Caprini evaluation OR^ =2.97, 95% CI (1.39, 6.32). Conclusions In this study, VTEs occurred infrequently. Considering the serious result of VTEs, it is important to prevent it. Age and Caprini are associated with elevated risk of postoperative VTEs. It is essential to target those at high risk for VTEs and appropriately treat those patients.
4.Influence of alprostadil on renal function, oxidative stress, and immune function in early diabetic nephropathy patients
Chao LIU ; Yan LIU ; Qiang LI ; Weili WU ; Qing HU
Journal of Chinese Physician 2017;19(3):396-398,402
Objective To investigate the influence of alprostadil on renal function,oxidative stress,and immune function in early diabetic nephropathy patients.Methods Eighty-six cases of early diabetic nephropathy patients in our hospital during April 2013 to January 2016 were chosen as research subjects,all patients were divided into observation (n =43) and control (n =43) groups according to random data form.Patients in observation group received alprostadil combined with pioglitazone treatment and those in control group received pioglitazone treatment only,both lasted for one month.Before treatment,one month after treatment,renal function indices were detected;serum oxidative stress indices were detected by automatic biochemical analyzer;T lymphocyte subsets in peripheral blood were measured by flow cytometry.Results Before treatment,there were no significant differences in renal function,oxidative stress,immune function between two groups (P > 0.05).One month after treatment,peripheral blood renal function indices such as blood urea nitrogen (BUN),estimated glomerular filtration rate (eGFR),and cystatin C (CysC) levels were lower in observation group than in control group,the level of serum albumin (Alb) was higher in observation group than in control group,the difference was statistically significant (P < 0.05);semm oxidation indices malondialdehyde (MDA),and advanced oxidation protein products (AOPP) were lower in observation group than in control group,human antioxidant indices such as superoxide dismutase (SOD),and total antioxidant capacity (T-AOC) were higher in observation group than in control group,the difference was statistically significant (P < 0.05);peripheral blood CD3 +,CD4 + T lymphocytes and the ratio of CD4 +/CD8 + were higher in observation group than in control group,CD8 +T lymphocyte level was lower in observation group than in control group,the difference was statistically significant (P < 0.05).Conclusions Alprostadil can optimize the renal function and immune function in early diabetic nephropathy patients,reduce the degree of oxidative stress.
6.Effects of nano porous beta-tricalcium phosphate/collagen scaffold modified with human bone morphogenetic protein 2 gene on differentiation of MC3T3-E1 cell lines
Qiang RUAN ; Gang ZHAO ; Rui GUO ; Yue XIAO ; Chao LI
Chinese Journal of Tissue Engineering Research 2016;20(38):5657-5663
BACKGROUND:Bone tissue transplantation or osteogenic material fil ing is after used for bone defect repair. To remove autologous bone tissues can lead to additional damage and secondary deformity, therefore, it is extremely urgent to search for a new osteogenic material. OBJECTIVE:To construct the porousβ-tricalcium phosphate (β-TCP)/col agen scaffold modified with human bone morphogenetic protein 2 (hBMP2) gene, and to observe its effects on differentiation of MC3T3-E1 cel lines. METHODS:The porousβ-TCP/col agen scaffold modified with hBMP2 gene was prepared. Then in vitro culture system of MC3T3-E1 cel lines with composite scaffold was established. There were scaffold and plate groups, and each group was divided into two subgroups according to the different concentrations of plasmid. Samples were col ected and observed morphological y by scanning electron microscope and light microscope after complex culture. After 1, 3, 7 and 14 days of induction, calcium nodules were observed through alizarin red staining, the cel cycle was detected by real-time PCR, and expressions ofαI-chain col agen type I gene, Osterix and bone sialoprotein were observed. RESULTS AND CONCLUSION:The number of cel s adhered, differentated and distributed on the composite scaffold was significantly higher than that of the single scaffold (P<0.05). Alizarin red staining and real-time PCR detection showed that the osteogenesis ability of MC3T3-E1 cel lines in the scaffold group was stronger than that in the plate group. To conclude, the porousβ-TCP/col agen scaffold modified with hBMP2 gene is an appropriate candidate for bone defect repair.
7.Expression of CXC chemokine receptor 4 in Barrett esophagus, esophageal adenocarcinoma and esophageal squamous cell carcinoma and its clinical significance
Chao ZHANG ; Hui TANG ; Lin LI ; Xinmin YAN ; Qiang GUO
Chinese Journal of Digestive Endoscopy 2010;27(11):589-593
Objective To investigate the expressions of CXCR4 in Barrett esophagus (BE), esophageal adenocarcinoma (EADC) and esophageal squamous cell carcinoma (ESCC), and its relationship with pathology, clinical staging and lymph node metastasis. Methods The expressions of CXCR4 in 56 cases of normal esophageal mucosa, 80 BE (including 22 BE with multifocal dysplasia), 25 EADC and 48 ESCC were examined with immunohistochemical method. Results CXCR4 was expressed in most samples of BE (80. 8% ), EADC (68. 0% ) and ESCC (78.4%) without significant difference ( P > 0. 05 ), which was significantly higher than that in normal esophageal mucosa (39. 3%, P <0. 01 ). The level of CXCR4 expression in BE, EADC or ESCC were not related with gender, age, or location of the foci ( P > 0. 05). There was no significant difference in CXCR4 expression between BE without dysplasia or BE with multifocal dysplasia ( P > 0. 05 ). CXCR4 expression level in well-differentiated EADC was significantly higher than that of mild or poorly differentiated (P < 0. 05 ). CXCR4 expression level was higher in EADC with lymph node metastasis than those without ( P < 0. 05 ). CXCR4 level in ESCC with TNM staging grades Ⅲ -Ⅳ was higher than that of grades Ⅰ - Ⅱ, and this variable was also higher in cases with lymph node metastasis than those without (P < 0. 05), so was the case of well and poorly differentiated ESCC (P < 0. 01 ). Conclusion Increased expression level of CXCR4 may be a common feature of EADC and ESCC, which is irrelevant to pathological types. CXCR4 level rises at the stage of BE, which is associated with the degree of tumor differentiation, lymph node metastasis and TNM staging. CXCR4 expression is of guiding significance in the diagnosis of BE, EADC and ESCC, and is the potential drug target.
8.K-ras mutations in colorectal cancer at different stages
Yongqi SHEN ; Yunbin YE ; Xiongwei ZHENG ; Chao LI ; Qiang CHEN
Tumor 2010;(2):134-137
Objective:To study the alteration of K-ras mutations in different stages of colorectal cancer(CRC) and its influence on the progression of CRC. Methods:The 20 paraffin-embedded tissues, including primary foci, metastatic lymph nodes, remoter metastatic foci, colorectal adenoma, and normal colorectal tissues, were collected from 20 patients with colorectal cancer. The sequence of PCR-amplified products were analyzed. Results:The wild K-ras gene was expressed in normal colorectal tissues. The mutation frequency of K-ras gene was 20.0% (4/20) in colorectal adenoma, 30.0% (6/20) in primary foci, 25.0% (5/20) in metastatic lymph nodes, and 30% (6/20) in remote metastatic lesions. In the samples with K-ras mutations, the consistency of the types of K-ras mutations between primary foci and colorectal carcinoma, lymph node metastatic lesions, remote metastatic lesions was 0.0%(0/4), 40.0%(2/5), and 50.0%(3/6), respectively.Conclusion:The colorectal adenoma, metastatic lymph nodes and remote metastatic lesions were not suited for K-ras analysis as routine samples in clinical practice. If the samples of primary lesions were not available, the detection results of metastatic lymph nodes and remote metastatic remote lesions will provide some reference values. K-ras gene had several different mutations in the progression of CRC.
9.Effects of the intensity of lower limb training after stroke
Nana FENG ; Qiang WANG ; Lin LI ; Chao HAN
Chinese Journal of Physical Medicine and Rehabilitation 2013;(4):290-294
Objective To investigate the effects of lower limb training at different intensities on the recovery of walking function after stroke.Methods Thirty-six stroke patients were randomly divided into a 40-minutes of training group,an 80-minutes of training group and a 120-minutes of training group.Because of three missing followup cases,the final numbers of cases were 12,11 and 10 cases respectively.All of the patients received lower limb training based on regular rehabilitation therapy,but at the three different intensities:40 minutes,80 minutes or 120 minutes a day,five days per week,for 4 weeks.Walking function was assessed using the Holden functional ambulation categories (FACs) and the Fugl-Meyer assessment (FMA).These were carried out before treatment,and after two and four weeks of treatment.Results Before treatment,the average FACs of the three groups were (1.83 ±0.94),(1.73 ± 1.01) and (1.80 ± 1.03) respectively.Their average FMA scores were (19.17 ±5.52),(23.00±4.71) and (19.40 ±7.90).After two weeks of treatment,the average FAC in the 120-minutes training group was (3.30 ± 0.48),significantly higher than in the 40-minutes training group.After four weeks the average FACs in the 40-minutes,80-minutes and 120-minutes training groups were (2.67 ± 0.65),(3.18 ± 0.60) and (3.80 ±0.42) respectively.The differences between the 120-minutes group and the 80-minutes group as well as between the 80-minutes group and the 40-minutes training group were statistically significant.The average FMA scores of the three groups after four weeks were (25.08±4.46),(28.64±3.56) and (25.90±5.19) respectively.All the differences were significant compared with pre-treatment.There were no significant differences in FMA scores among the three groups after two weeks or four weeks of treatment.Before treatment,the proportion of patients able to walk independently in the 40-minutes,80-minutes and 120-minutes training groups were 16.7%,18.2% and 20.0%.After two weeks the proportions had risen to 33.3%,36.4% and 100% respectively,so the results in the 120-minutes training group were significantly better.After four weeks of treatment the proportions of the three groups were 58.3%,90.9% and 100%.All these improvements were significant compared with pre-treatment,but when compared with the 80-minutes training group,neither of the others showed a significant difference.Conclusions Intensive training can accelerate the recovery of walking function of patients after stroke and promote their ability to participate in daily activities.
10.Relationship between the functional single nucleotide polymorphism rs1052133 in the 8-oxoguanine DNA glycosylase 1 (OGG1) gene and vitiligo in a Chinese Han population
Chao WEI ; Huini QIANG ; Tianwen GAO ; Chunying LI
Chinese Journal of Dermatology 2013;(4):227-230
Objective To estimate the relationship of the functional single nucleotide polymorphism (SNP) rs1052133 in the 8-oxoguanine DNA glycosylase 1 (OGG1) gene with vitiligo in a Chinese Han population.Methods Blood samples were collected from 800 patients with vitiligo and 800 healthy human controls,and subjected to genomic DNA extraction.PCR-restriction fragment length polymorphism (PCR-RFLP) analysis was performed to analyze the genotype of the SNP rs1052133 in the OGG1 gene.The relationship between the SNP and the risk of vitiligo was evaluated by chi-square test and unconditional logistic regression analysis.Enzyme linked immunosorbent assay (ELISA) was carried out to assess the serum level of 8-hydroxydeoxyguanosine (8-OHdG) in 83 patients with vitiligo and 83 healthy human controls,then,t test was used to compare the serum 8-OHdG level between the patients and controls.Results The frequency of CC,CG and GG genotype of the SNP rs1052133 was 16.8%,54.0% and 29.2% respectively in the patients,21.4%,52.8% and 25.8%respectively in the controls (x2 =6.26,P < 0.05).Increased frequency of G allele of the SNP rs1052133 was observed in the patients with vitiligo compared with the controls (56.2% vs.52.2%,x2 =5.16,P < 0.05).A statistically increased risk of vitiligo was associated with the CG (x2 =3.98,P < 0.05,adjusted odds ratio 1.31,95% confidence interval:1.01-1.70) and GG (x2 =6.01,P < 0.05,adjusted odds ratio 1.45,95% confidence interval:1.08-1.94) genotype of SNP rs1052133 compared with the CC genotype,which was more evident among the patients with the following characteristics:female,nonsegmental vitiligo,active vitiligo,long clinical course (> 12 months),a family history of vitiligo,and no accompanied autoimmune diseases.In addition,the patients with the CG or GG genotype of SNP rs1052133 had a higher serum 8-OHdG level than those with the CC genotype ((838.23 ± 294.11) μg/L vs.(593.84 ± 190.14) μg/L,t =3.63,P < 0.01).Conclusions The SNP rs1052133 in the OGG1 gene may be responsible for the development of vitiligo in Chinese Han populations,which is likely to be associated with defects in DNA repair.