1.Correlation between connexin37 C1019T polymorphism and ischemic stroke and its outcome
Dan LIU ; Jiangong SUN ; Hongying SUN ; Guangwei ZHANG ; Jia ZHANG ; Guixi WANG ; Jing YANG ; Ruiqi SONG
International Journal of Cerebrovascular Diseases 2012;20(4):251-256
Objective To investigate the correlation between cornexin37 (Cx37) CI019T polymorphism and ischemic stroke and its outcome.Methods Restriction fragment length polymorphism analysis was used to detect the distribution of Cx37 C1019T polymorphism in a ischemic stroke group and a control group.The modified Rankin scale (mRS) was used to evaluate the neurological outcome at 3 months after onset.Results A total of 235 patients in the control group,and 232 patients in the ischemic stroke goup were recruited.In the ischemic stroke group,210 had a good outcome (mRS <3) and 22 had a poor outcome (mRS≥ 3).The TT genotype (12.93% vs.6.39% ; x2 =10.087,P =0.006) and T allele (31.25% vs.21.49% ; x2 =11.466,P=0.001) frequency in the ischemic stroke group were significantly higher than those in the control group.Multivariatelogistic regression analysis showed that TT genotype (odds ratio [OR] 5.794; 95% confidence interval [CI] 1.405-23.894; P =0.015) and T allele (OR 131.016,95% CI 6.943 -2 472.477; P =0.001)signifkantly increased the risk of ischemic stroke.Univariate analysis showed that TT genotype (OR 0.650,95% CI 0.144 - 2,934; P =0.575),CT genotype (OR 0.622,95% CI 0.234 - 1.655; P =0.342),and CC genotype (OR 0.654,95% CI 0.268 - 1.595; P =0.350) had no significant correlation with the outcome of ischemic stroke.Conclusions Cx37 1019TT genotype and T allele may increase the risk of ischemic stroke.T allele is one of genetic susceptibility factors for ischemic stroke; however,its gene polymorphism is not associated with the outcome of ischemic stroke at 3 months after onset.
2.Association between codon 54 polymorphism of intestinal fatty acid-binding protein 2 gene and plasma lipids in middle-aged and old populations
Xiaotong CHANG ; Zhenhui WANG ; Lijuan HOU ; Biaoying LI ; Minggang DONG ; Guixi LI
Chinese Journal of Tissue Engineering Research 2008;12(7):1397-1400
BACKGROUND: Studies have shown that alanine (A) to threonine (T) substitution at codon 54 of intestinal fatty acid-binding protein (FABP2) in different populations is associated with dyslipidemia and other characteristics of metabolic syndrome.OBJECTIVE: To investigate the frequency of encoding 54Ala/Thr (A/T) single nucleotide polymorphism in the FABP2 in middle-aged and old people, and explore the association between 54T FABP2 and plasma lipids.DESIGN: A case-controlled analysis. SETTING: Department of Biochemistry, Hebei North University and Department of Clinical Laboratory, the 251 Hospital of Chinese PLA.PARTICIPANTS: 469 physical examinees were selected from the Medical Examination Center, the 251 Hospital of Chinese PLA between October 2003 and April 2005. The subjects included 217 males with mean age of (56±10) years, and 252 females with mean age of (55±13) years. Only people with normal liver and kidney function, and with no blood relation were recruited. The informed consent to this study was obtained from all subjects. The experiment was admitted by Hospital Ethics Committee. METHODS: ①After fasting for 12 hours, automatic analyzer (Olympus AU 6400) was adopted to measure plasma total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C), apoprotein A1(Apo A1) and Apo B levels. ②1 mL venous blood was extracted and immediately mixed with anti-coagulants containing citric acid, natrium citricum and glucose. White blood cells were separated and genomic DNA was isolated using standard methods with proteinase K digestion and phenol/chloroform purification. The genotype distribution frequency in each group was detected by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). MAIN OUTCOME MEASURES: ①Plasma TC, TG, HDL-C, LDL-C, Apo A1and Apo B levels; ②Distributions of FABP2 genotypes at codon 54. RESULTS: ①The genotype frequencies of A/A, A/T, T/T were 0.48, 0.42, and 0.10 in males, and 0.44, 0.46, and 0.10 in females, respectively. The allelic frequency of point mutant 54Thr in FABP2 gene was 0.31 in males and 0.33 in females, respectively. There was no difference between males and females (χ2=0.47, P > 0.05). ②The LDL-C and Apo B concentrations in fasting plasma of males with 54T allele were significantly higher than those with 54A allele (P < 0.05). The TC and LDL-C concentrations in fasting plasma of females with 54T allele were significantly higher than those with 54A allele (P < 0.05). CONCLUSION: In the middle-aged and old populations, the frequency of encoding 54Ala/Thr polymorphism in FABP2 gene is not correlated with gender, but with high lipoprotein profile.
3.A study on the serum proteomics in patients of colorectal cancer
Xiaomei DENG ; Guixi ZHENG ; Chuanxin WANG ; Jian ZHANG ; Wei LI ; Xiao ZHANG
Chinese Journal of General Surgery 2008;23(9):713-715
Objective To detect biomarker proteins in relation to metastasis by comparing serum protein profiles of primary colorectal cancer patients with or without metastasis. Methods A total of 219 serum samples were analyzed using surface enhanced laser desorption ionization time of flight mass spectrometry(SELDI-TOF-MS).The samples were divided into two groups:the training group consisting of 57 primary colorectal cancer patients、63 metastatic colorectal cancer patients and 42 healthy controls,and the test group consisting of 26 primary colorectal cancer patients and 31 metastatic colorectal cancer patients.Samples in training group were analyzed to identify serum biomarker proteins which could differentiate colorectal cancer patients with or without metastasis.The sensitivity and specificity of biomarker proteins were examined by results from blind test group. Results In the m/z region of 2000~30000,31 proteins had statistically significant difference between primary colorectal cancer patients and healthy controls.The m/z of difierentiated proteins were respectively 3240.7、9289.3、5334.4、4596.1 and 4792.4 according to P value.38 proteins had statistically significant diffefence between metastatic colorectal cancer patients and healthy controls.The m/z of differentiated proteins were respectively 3240.7、9289.3、9184.4、3191.5 and 9340.9 according to P value.Only two protein peaks(9184.4 and 9340.9)were found of statistical difference between primary eolorectal cancer patients and metastatic colorectal cancer patients.The sensitivity and specificity of the combination use of the two biomarkers were respectively 90.3% and 88.5% in the test group. Conclusion SELDI-TOF-MS was helpful to find protein biomarkers with relation to metastasis in colorectal carcinoma patients.
4.Establishment of detection method for serum circulating Bmi-1 mRNA and its application in diagnosis of colorectal cancer
Xin ZHANG ; Haiyan WANG ; Guixi ZHENG ; Lili WANG ; Peilong LI ; Tong LIU ; Yongmei YANG ; Lutao DU ; Juan LI ; Chuanxin WANG
Chinese Journal of Laboratory Medicine 2014;(9):678-682
Objective To establish a direct reverse transcription real-time fluorescence quantitative polymerase chain reaction ( RT-qPCR-D ) method for detecting serum circulating B cell-specific moloney murine leukemia virus integration site-1 (Bmi-1) mRNA, and analyze the levels of serum circulating Bmi-1 mRNA in colorectal cancer patients by using of this method for exploring its diagnosis value in colorectal cancer.Methods Methodology establishment.RNA was extracted from colorectal cancer HT 29 cell line, and detection standard curves of Bmi-1, ubiquitin C ( UBC), glyceraldehyde-3-phosphate dehydrogenase ( GAPDH) mRNAs were established , then the amplification efficiencies were calculated.Bmi-1 mRNA level was directly detected in serum and preparation buffer mixture , then the specificity of assay was evaluated by melting curve, and detection limit was observed through diluted serum samples.The serum circulating Bmi-1 mRNA levels were detected by ELISA in 158 cases with colorectal cancer , of which there were 26 cases of tumor node metastasis ( TNM)Ⅰstage, 53 cases of TNMⅡ, 47 cases of TNMⅢ, 32 cases of TNMⅣand 53 cases of controls with normal colonoscopy collected from January 2008 to January 2009 in Qilu Hospital of Shandong University.Comparisons of groups were determined by applying Mann-Whitney U test or Kruskal-Wallis test, and receiver operating characteristic ( ROC) curves were established to illustrate the diagnostic performance.Results The log values of Bmi-1, UBC and GAPDH showed good linear correlations with quantification cycle (Cq) values(R2 =0.990, 0.990, 0.991, all P <0.001), and the amplification efficiencies were 0.875, 0.917 and 0.935, respectively.Using the established RT-qPCR-D method, the peak of melting curve of Bmi-1, UBC and GAPDH mRNAs were single, the detection limit was up to 1.25μl.The levels of serum circulating Bmi-1 mRNA detected by RT-qPCR-D were 0.138 ( 0.078-0.228 ) in colorectal cancer stage Ⅰ patients, 0.163(0.067 -0.287) instage Ⅱ patients, 0.217(0.072-0.267) instage Ⅲpatients, 0.273(0.139 -0.419) in stage Ⅳ patients and 0.021(0.008 -0.029) in health controls, a significant difference was found among groups ( H =89.5, P <0.001 ).The levels of serum circulating Bmi-1 mRNA in each stage colorectal cancer were all significantly higher than that in control group(U=58.0, 287, 246, 72.5,all P<0.001).The levels in Ⅳstage patients were significantly higher than those in other stages patients (U=247, 590, 540,P=0.008, 0.020, 0.035), while no significant differences among Ⅰstage,Ⅱstage and Ⅲstage patients(U=633, 514, 1170,all P>0.05).ROC curve analysis showed area under the ROC curve ( AUC) for serum circulating Bmi-1 mRNA was 0.921(95%CI=0.876-0.953), which was significantly superior to the AUC of CEA (0.745, 95%CI=0.680-0.802, Z=4.697, P<0.001 ).When cutoff value was 0.034, the diagnostic sensitivity and specificity was 89.2%(141/158) and 90.6%(48/53), while 41.8%(66/158) and 73.6%(39/53) using CEA.The AUC for combination of circulating Bmi-1 mRNA and CEA was 0.933(95%CI=0.890-0.963 ) , which was no statistical significance when compared with the AUC of circulating Bmi-1 mRNA(Z=4.697, P>0.05).Conclusions The study establishes a higher sensitive, specific for detecting serum circulating Bmi-1 mRNA. Based on this method , serum circulating Bmi-1 mRNA is found to be increased in colorectal cancer , and is superior to traditional tumor marker CEA in diagnosis of colorectal cancer, which may become a potential detection index for early detection of colorectal cancer.
5.Percutaneous transluminal angioplasty for the treatment of diabetic foot
Wenduo ZHANG ; Guixi JIANG ; Qiang LI ; Zhijun XUE ; Jinping GUAN ; Jin MIAO ; Wenxian HU ; Juanzi ZHANG ; Jinjun WANG
Chinese Journal of General Surgery 2011;26(7):573-576
Objective To evaluate percutaneous transluminal angioplasty ( PTA) for stenosed arteries of the lower extremities in patients with ischaemic diabetic foot. Methods We retrospectively analyzed the clinical and follow-up data of using PTA to treat diseased infrapopliteal arteries in diabetic patients who were hospitalized from Oct,2006 to May,2008. Results Technical success rate was 87% , procedure related complications developed in 8. 9% of patients, postoperative complications were 11. 1% , perioperative mortality was 2. 5% , limb salvage rate was 90% , pain symptom was significantly mitigated or relieved, ulcer healed well. The median hospitalstay was 10 days. Restenosis rates were 38. 1 % , 50% respectively at 1 year and 2 years. Rest pain and ulcer recurrence rates were 10% and 12% at 1 year and 2 years respectively; Amputation rates were 10% and 15. 3% at 1 year and 2 years. Restenosis ( or occlusion) , rest pain or ulcer recurrence and amputation rate in Fontain Ⅳ group is significantly poorer than that in Fontain Ⅰ - Ⅲ group (P <0. 05). Conclusions Percutaneous transluminal angioplasty (PTA) for critical limb ischeamia in patients with ischaemic diabetic foot are feasible, with minimal invasiveness, low complications. Fontain classification predicts PTA thrapeutic results.
6.Effect of Miao Nationality Herbs Spray for Serum SOD, MDA and Expression of Fas-mRNA and Caspase-3 mRNA in Lung Tissues of Silica-treated Rats
Gao HUANG ; Guangzhi HE ; Guixi SONG ; Xia LIU ; Feng CAO ; Wenjia WANG ; Yuchen LIU ; Changfu YANG ; Kun CAI ; Bangxiang XU ; Chuan QIN ; Jinhuo WAN ; Zhenlian LUO ; Jianmei HUANG
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(5):1172-1177
This study was aimed to prepare the spraying agent of prescriptions of Miao nationality herb and investigate the effect of Miao nationality herbs spray for serum SOD, MDA, and expression of Fas and Caspase-3 mRNA in lung tissues of silica-treated rats. The healthy SD rats were divided into 5 groups. Silica dust suspension was used in the model establishment of 4 groups. After the model was successfully established, 3 groups were randomly selected and given glucocorticoids atomization inhalation, Miao nationality herbs spray, Miao nationality herbs spray combined with intragastric administration of herbal medicine, respectively. After 40-day treatment, water-solubletetrazolium salt (WST-1) was used in the detection of serum superoxide dismutase (SOD). Thiobarbituric acid (TBA) was used in the detection of malondialdehyde (MDA). The mRNA expression variance of the Fas and Caspase-3 were detected by RT-PCR. The results showed that compared with the silica dust suspension group, the SOD activity of serum in the Miao nationality herbs spray group was significantly increased (P< 0.05). MDA content and the mRNA of Fas and Caspase-3 were significantly lower in the Miao nationality herbs spray group (P< 0.05). It was concluded that Miao nationality herbs spray group was able to increase the SOD activity of serum, decrease MDA content, and obviously decrease the expression of Fas and Caspase-3 of lung tissues among silica dust suspension rats.
7.Regulation Mechanism of Occludin Function and Factors Affecting Its Expression
Shi-Jia ZHANG ; Juan-Xia ZHENG ; Cheng-Wei WANG
Progress in Biochemistry and Biophysics 2024;51(11):2832-2844
Tight-junction (TJ) is a complex supramolecular entity composed of complete membrane proteins, membranes and soluble cytoplasmic proteins, which is distributed in almost all barrier structures in the body. It can maintain the polarity of epithelial cells, close the intercellular space and prevent the overflow of materials in the epithelial space, and is a highly dynamic signaling entity. Occludin is one of the most representative members of TJ proteins, mainly responsible for sealing intercellular connections, maintaining intercellular permeability, and participating in maintaining the integrity of vascular endothelium. The integrity of occludin is related to the integrity of TJ, and the function of occludin is often associated with the barrier properties of various tissues, and the abnormal expression of occludin is related to the occurrence and development of various diseases. Occludin contains abundant Ser and Thr residues and has multiple phosphorylation sites. Phosphorylation is necessary for the combination of occludin and TJ, which can regulate the location of occludin, regulate the expression of occludin, and enhance the permeability and barrier function of TJ. Therefore, phosphorylation regulation is a mechanism that cannot be ignored in the regulation of occludin function. Occludin also interacts with many other proteins, such as co-forming the cytoskeleton with ZO-1, and is regulated by a variety of transcription factors. Studies have confirmed that in pathological conditions, a variety of signaling pathways can disrupt the integrity of cell barrier by regulating the expression and distribution of occludin. Myosin light chain kinase (MLCK) signal transduction pathway is one of the important ways to regulate the structure and function of TJ. It influences the expression of occludin by altering the cytoskeleton. MLCK mainly uses the phosphorylation of myosin light chain (MLC) as a medium to promote actin contraction, secondary decomposition of tightly binding proteins, resulting in increased or changed cellular barrier permeability, and increased MLC phosphorylation is also a biochemical marker of actomyosin contraction. Activation of MLCK causes Thr18 and Ser19 phosphorylation of MLC, which promotes the assembly of myosin II into myosin fibers and activates the hydrolysis of ATP, which relaxes the intercellular connections and reduces the ability of upper cortex to resist external invaders. Protein kinase C (PKC) plays an important role in the regulation of tightly connected signaling molecules, affecting the dynamic changes of paracellular permeability. PKC pathway is a key link in many cell signal transduction pathways, which influences all aspects of cell activities by catalyzing Ser/Thr residues phosphorylation of membrane proteins and many enzyme proteins. After PKC activation, it can regulate cellular barrier function by phosphorylating occludin and inducing its redistribution, and directly affect TJ action. Specific PKC subunits such as PKCα, PKCδ and PKCγ are activated and act on occludin molecules to promote their phosphorylation and cause the increase of TEER. The increase of TEER helps to regulate intercellular TJ and enhance the tightness of intercellular connections. Mitogen-activated protein kinases (MAPK) are usually activated by inflammatory factors, during which different signal transduction pathway subfamilies are formed to regulate occludin expression and affect tight junction and mucosal barrier functional integrity. Meanwhile, occludin is easily affected by various factors (such as cytokines and flora toxins), and abnormal expression of occludin will lead to structural damage of TJ and further damage of the intercellular barrier. Therefore, this paper summarizes the molecular structure and physiological function of occludin, and further summarizes its related signal regulation pathways and influencing factors, in order to provide theoretical support for maintaining the integrity of barrier function of occludin.
8.Optimization of the Extraction Technology of Total Flavonoids from Typhonium divaricatum
Mengmeng WANG ; Dian XIE ; Yufang ZHOU ; Xiaojing LI ; Bubin XU ; Bin ZHOU
China Pharmacy 2019;30(14):1949-1952
OBJECTIVE: To establish a method for the content determination of total flavonoids from Typhonium divaricatum, and to optimize its extraction technology. METHODS: The content of total flavonoids in T. divaricatum was determined by UV spectrophotometry. Using the extraction amount of total flavonoids from T. divaricatum as index, the volume fraction of ethanol, the ratio of material to liquid, the extraction time and times as factors, the extraction technology of total flavonoids from T. divaricatum was optimized by L9(34) orthogonal design, based on the single factor test. RESULTS: The linear range of rutin were 8-48 μg/mL (r=0.999 7); the quantification limit was 0.54 μg/mL, and the detection limit was 0.18 μg/mL; RSDs of precision, stability and repeatability tests were all lower than 2%. The recoveries were 99.61%-102.38%(RSD=1.15%, n=6). The optimal extraction technology was as follows: ethanol concentration of 70%, solid-liquid ratio of 1 ∶ 20 (g/mL), extraction time of 45 min, extracting for 2 times. Under this condition, the average content of total flavonoids from T. divaricatum was 2.74 mg/g. CONCLUSIONS: Established method is simple and accurate; the extraction process is stable and feasible.
9. Follow-up of people living with HIV/AIDS by primary health care institutions in rural area of Jiangxi province
Pengfei FAN ; Qing YANG ; Yurong MAO ; Qiang HU ; Houlin TANG ; Jian LI ; Yaling LUO ; Fen WANG ; Huanqing ZHAN ; Siming ZANG
Chinese Journal of Epidemiology 2019;40(3):346-349
Objective:
To understand the current status of follow up of people living with HIV/AIDS by health service at grass root in rural area of Jiangxi province and related factors, and provide references for the promotion of the follow up by grass root health service.
Methods:
People living HIV/AIDS aged ≥18 years and diagnosed before 31 December 2017 in 6 townships of Xinjian, Yushan counties and Guixi city were included in the study in Jiangxi province. They had been followed up for more than one time after the first epidemiologic survey. The information about their demographic characteristics and HIV infection status were collected by using self-designed questionnaire. Univariate and multivariate logistic regression analyses were conducted to identify the factors that influencing the acceptance of follow up by grass root health service.
Results:
Of the 373 surveyed HIV infected subjects aged (53.06±16.15) years, 261 were males (70.0
10. The situation and related factors of satisfaction of HIV-infected people towards the follow-up management of primary medical and health care institutions in rural areas of Jiangxi Province
Pengfei FAN ; Qing YANG ; Yurong MAO ; Houlin TANG ; Jian LI ; Fen WANG ; Huanqing ZHAN ; Siming ZANG
Chinese Journal of Preventive Medicine 2019;53(5):522-526
The HIV-infected people were investigated for their satisfactory situation towards the follow-up management of primary medical and health care institutions in Xinjian County, Yushan County and Guixi City of Jiangxi Province from January to July 2018 and related factors were also analyzed. The total score of the Infected Patients′ Follow-up Management Satisfaction Scale higher than 59 was defined as the satisfactory. 75.68% of 259 infected patients were satisfied with the follow-up management. Compared to farmers and other follow-up modes, non-farmers (