1.Application of dyes in polyacrylamide upper gel electrophoresis and im-munoblotting
Weiguang CHEN ; Chenjie YAN ; Zhaojin XU ; Yiting DONG ; Zhichao MAO ; Jiawei SUN ; Wantie WANG ; Linbo YUAN
Chinese Journal of Pathophysiology 2025;41(5):1036-1040
AIM:To investigate the effect of dyes,Remazol BrOrange yellow(RBY)and erythrosine(ERY),on the outcomes of immunoblotting analysis when used for staining the concentrate gel in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE).METHODS:Polyacrylamide gels were divided into five groups:the control group(prepared according to the conventional kit protocol),the RBY-stained group with a final concentration of 0.08 g/L,the RBY-stained group with a final concentration of 0.16 g/L,the ERY-stained group with a final concentration of 0.08 g/L,and the ERY-stained group with a final concentration of 0.8 g/L.Gels were prepared and subjected to electro-phoresis,followed by coomassie brilliant blue staining to visualize protein bands.Subsequently,proteins were transferred to PVDF membranes,which were then blocked,incubated with primary and secondary antibodies,washed,and finally ex-posed for imaging to observe the target protein vinculin bands.RESULTS:Compared with the unstained concentrate gel,the loading wells of the RBY or ERY pre-stained concentrate gel were more clearly visible.Analysis of the gels stained with coomassie brilliant blue after electrophoresis and marker visualization showed no significant different in protein elec-trophoretic mobility between prestained and unstained gels.Comparative analysis of the immunoblotting also indicated that the detection of protein samples transferred to PVDF membranes was unaffected.CONCLUSION:Prestaining concen-trate gels with RBY or ERY can enhance the efficiency of gel-based electrophoresis and immunoblotting analysis.
2.Clinical study on high-dose ilaprazole combined with amoxicillin for newly diagnosed elderly patients with Helicobacter pylori infection
Chen LI ; Yujing WANG ; Jianna MAO ; Hao GUO ; Yuhou SHEN ; Zhichao DONG ; Binbin YAN
China Pharmacy 2025;36(14):1792-1796
OBJECTIVE To evaluate the efficacy and safety of high-dose ilaprazole combined with amoxicillin for newly diagnosed elderly patients with Helicobacter pylori (Hp) infection, and analyze independent risk factors for failure of Hp infection eradication treatment. METHODS Totally 200 cases of newly diagnosed elderly patients with Hp infection in Xinxiang Central Hospital from August 1, 2021 to December 1, 2024 were selected and randomly divided into control group and study group, with 100 cases in each group. The control group was treated with classic quadruple therapy regimen (Amoxicillin capsules+ Clarithromycin tablets+Bismuth potassium citrate tablets+Ilaprazole enteric-coated tablets). The study group was treated with high- dose Ilaprazole enteric-coated tablets+Amoxicillin capsules. All patients were administered medication for 2 weeks. Hp eradication rates in the two groups were compared using intention-to-treat (ITT) and per-protocol (PP) analyses. The incidence of adverse reactions in both groups was also recorded. The multiple-factor Logistic regression analysis was used to identify independent risk factors for failure of Hp infection eradication treatment. RESULTS In ITT and PP analyses, there was no significant difference of Hp eradication rates between the two groups (P>0.05). There was no significant difference in incidence of mild to moderate adverse reactions between the two groups (P>0.05). BMI ≤18.5 kg/m2, BMI >23.9 kg/m2, rural residence, concomitant diabetes and concomitant heart disease were identified as independent risk factors influencing the failure of Hp infection eradication treatment (P<0.05). CONCLUSIONS The efficacy and safety of high-dose ilaprazole combined with amoxicillin are comparable to classic quadruple therapy regimen in treating newly diagnosed elderly patients with Hp infection. Independent risk factors influencing the failure of Hp infection eradication treatment include BMI ≤18.5 kg/m2, BMI >23.9 kg/m2, rural residence, concomitant diabetes and concomitant heart disease.
3.Expression and clinical significance of serum CXCL1 and PRDM5 in lymph node metastasis of progressive gastric cancer
Xiaoying DING ; Zhichao DONG ; Jianna MAO ; Changqing GUO ; Aimin YUE
The Journal of Practical Medicine 2025;41(20):3206-3213
Objective To investigate the relationship between serum CXC chemokine ligand 1(CXCL1)and positive regulatory zone zinc finger protein 5(PRDM5)levels and lymph node metastasis of progressive gastric cancer and to analyze their predictive value for patients'prognosis.Methods 203 patients with progressive gastric cancer diagnosed in our hospital from June 2020 to March 2023 were selected and divided into the lymph node metastasis group(n=90)and the no-lymph node metastasis group(n=113)based on the presence or absence of lymph node metastasis,and the differences in the general information of the two groups were analyzed and compared,and the diagnostic value of CXCL1 and PRDM5 in lymph node metastasis of progressive gastric cancer was analyzed by plotting the ROC curve.Logistic regression was used to analyze the risk factors of lymph node metastasis in patients with progressive gastric cancer.Follow up for 2 years,draw Kapan Meier curves to compare the prognosis of patients with advanced gastric cancer lymph node metastasis at different levels of CXCL1 and PRDM5.Results The CXCL1 level in the lymph node metastasis group was higher than that in the no-lymph node metastasis group,and its PRDM5 level was lower than that in the no-lymph node metastasis group(P<0.05).The AUCs for diagnosing lymph node metastasis of progressed gastric cancer were 0.755 and 0.844 for CXCL1 and PRDM5,respectively,and the AUC for the combination of the two was 0.898(95%CI 0.848~0.936).The sensitivity and specificity were 88.89%and 77.88%,respectively(P<0.05).Tumor size,differentation degree,serum CEA,serum CA19-9,CXCL1,and PRDM5 levels were all risk factors for lymph node metastasis in patients with progressive gastric cancer(P<0.05).The survival time of patients with CXCL1>96.13 pg/mL is(15.13±0.85)months,while the survival time of patients with CXCL1≤96.13 pg/mL is(19.06±0.66)months.The survival time of patients with CXCL1≤96.13 pg/mL is longer than that of patients with CXCL1>96.13 pg/mL(P<0.05).The survival time of patients with PRDM>100.85 pg/mL is(18.62±0.69)months,while the survival time of patients with PRDM≤100.85 pg/mL is(14.60±0.78)months.The survival time of patients with PRDM>100.85 pg/mL is longer than that of patients with PRDM≤100.85 pg/mL(P<0.05).Conclusion The abnormal expression of CXCL1 and PRDM5 is related to lymph node metastasis in patients with progressive gastric cancer,and the combined detection of the two is of high value in the assessment of lymph node metastasis and prognosis in patients with progressive gastric cancer.
4.Expression and clinical significance of serum CXCL1 and PRDM5 in lymph node metastasis of progressive gastric cancer
Xiaoying DING ; Zhichao DONG ; Jianna MAO ; Changqing GUO ; Aimin YUE
The Journal of Practical Medicine 2025;41(20):3206-3213
Objective To investigate the relationship between serum CXC chemokine ligand 1(CXCL1)and positive regulatory zone zinc finger protein 5(PRDM5)levels and lymph node metastasis of progressive gastric cancer and to analyze their predictive value for patients'prognosis.Methods 203 patients with progressive gastric cancer diagnosed in our hospital from June 2020 to March 2023 were selected and divided into the lymph node metastasis group(n=90)and the no-lymph node metastasis group(n=113)based on the presence or absence of lymph node metastasis,and the differences in the general information of the two groups were analyzed and compared,and the diagnostic value of CXCL1 and PRDM5 in lymph node metastasis of progressive gastric cancer was analyzed by plotting the ROC curve.Logistic regression was used to analyze the risk factors of lymph node metastasis in patients with progressive gastric cancer.Follow up for 2 years,draw Kapan Meier curves to compare the prognosis of patients with advanced gastric cancer lymph node metastasis at different levels of CXCL1 and PRDM5.Results The CXCL1 level in the lymph node metastasis group was higher than that in the no-lymph node metastasis group,and its PRDM5 level was lower than that in the no-lymph node metastasis group(P<0.05).The AUCs for diagnosing lymph node metastasis of progressed gastric cancer were 0.755 and 0.844 for CXCL1 and PRDM5,respectively,and the AUC for the combination of the two was 0.898(95%CI 0.848~0.936).The sensitivity and specificity were 88.89%and 77.88%,respectively(P<0.05).Tumor size,differentation degree,serum CEA,serum CA19-9,CXCL1,and PRDM5 levels were all risk factors for lymph node metastasis in patients with progressive gastric cancer(P<0.05).The survival time of patients with CXCL1>96.13 pg/mL is(15.13±0.85)months,while the survival time of patients with CXCL1≤96.13 pg/mL is(19.06±0.66)months.The survival time of patients with CXCL1≤96.13 pg/mL is longer than that of patients with CXCL1>96.13 pg/mL(P<0.05).The survival time of patients with PRDM>100.85 pg/mL is(18.62±0.69)months,while the survival time of patients with PRDM≤100.85 pg/mL is(14.60±0.78)months.The survival time of patients with PRDM>100.85 pg/mL is longer than that of patients with PRDM≤100.85 pg/mL(P<0.05).Conclusion The abnormal expression of CXCL1 and PRDM5 is related to lymph node metastasis in patients with progressive gastric cancer,and the combined detection of the two is of high value in the assessment of lymph node metastasis and prognosis in patients with progressive gastric cancer.
5.Application of dyes in polyacrylamide upper gel electrophoresis and im-munoblotting
Weiguang CHEN ; Chenjie YAN ; Zhaojin XU ; Yiting DONG ; Zhichao MAO ; Jiawei SUN ; Wantie WANG ; Linbo YUAN
Chinese Journal of Pathophysiology 2025;41(5):1036-1040
AIM:To investigate the effect of dyes,Remazol BrOrange yellow(RBY)and erythrosine(ERY),on the outcomes of immunoblotting analysis when used for staining the concentrate gel in sodium dodecyl sulfate-polyacryl-amide gel electrophoresis(SDS-PAGE).METHODS:Polyacrylamide gels were divided into five groups:the control group(prepared according to the conventional kit protocol),the RBY-stained group with a final concentration of 0.08 g/L,the RBY-stained group with a final concentration of 0.16 g/L,the ERY-stained group with a final concentration of 0.08 g/L,and the ERY-stained group with a final concentration of 0.8 g/L.Gels were prepared and subjected to electro-phoresis,followed by coomassie brilliant blue staining to visualize protein bands.Subsequently,proteins were transferred to PVDF membranes,which were then blocked,incubated with primary and secondary antibodies,washed,and finally ex-posed for imaging to observe the target protein vinculin bands.RESULTS:Compared with the unstained concentrate gel,the loading wells of the RBY or ERY pre-stained concentrate gel were more clearly visible.Analysis of the gels stained with coomassie brilliant blue after electrophoresis and marker visualization showed no significant different in protein elec-trophoretic mobility between prestained and unstained gels.Comparative analysis of the immunoblotting also indicated that the detection of protein samples transferred to PVDF membranes was unaffected.CONCLUSION:Prestaining concen-trate gels with RBY or ERY can enhance the efficiency of gel-based electrophoresis and immunoblotting analysis.
6.Mesenchymal stem cells inhibit hypoxic damage to rat pulmonary microvascular endothelial cells by regulating oxidative stress
Zhuang MAO ; Xue LI ; Changyao WANG ; Lin LYU ; Hu CAO ; Zhichao HE ; Zuyin YU ; Hua WANG
Chinese Journal of Pharmacology and Toxicology 2024;38(7):504-510
OBJECTIVE To explore the role and mechanism of dental pulp stem cells(DPSCs)in repairing hypoxic injury to rats pulmonary microvascular endothelial cells(PMVECs).METHODS ①PMVECs were treated with cobalt chloride at 0,10,25,50 and 100 μmol·L-1 for 72 h.CCK-8 was used to detect the cell viability,and the protein levels of hypoxia-inducible factor 1α(HIF-1α),zona occludens small-band protein 1(ZO-1),and occludin(OCLN)were detected by Western blotting.②There was a cell control group,model group,and model+DPSCs group,and the levels of reactive oxygen species(ROS)was detected by immunofluorescence staining after at 24 and 48 h of action.The levels of ZO-1 and OCLN proteins were detected by Western blotting.③ A cell control group,model group,model+DPSC group and model+DPSC cell knockdown superoxide dismutase 1(SOD1)group were set up.The mRNA level of SOD1 was detected by real-time fluorescence quantitative PCR 24 and 48 h later,while the protein levels of ZO-1 and OCLN were detected by Western blotting.RESULTS ① Com-pared with the cell control group,72 h of cobalt chloride 100 μmol·L-1 treatment of PMVECs resulted in a cell survival rate above 80%,a significant increase in the level of HIF-1α protein(P<0.05),a signifi-cant decrease in the levels of ZO-1 and OCLN proteins(P<0.01),and establishment of a model of hypoxic injury in PMVECs.② Compared with the cell control group,the ROS level was significantly higher in the model group(P<0.01).Compared with the model group,the ROS level was significantly lower in the model+DPSCs group(P<0.01),while the levels of ZO-1 and OCLN proteins were signifi-cantly higher in the model+DPSCs group(P<0.05).③ Compared with the DPSC group,ZO-1 and OCLN expressions were significantly decreased after knockdown of SOD1 in DPSCs(P<0.05,P<0.01).CONCLUSIONS DPSCs can repair hypoxic injury to PMVECs,and the anti-oxidative stress capacity of DPSCs plays an important role in hypoxic injury repair of PMVECs.
7.Delay in identification, healthcare-seeking, and definitive diagnosis of tuberculosis among students in Urumqi City from 2010 to 2019
Li MA ; Zhichao LIANG ; Yanggui CHEN ; Weisheng ZHANG ; Hongkai MAO ; Wanting XU ; Mingqin CAO
Journal of Preventive Medicine 2023;35(1):53-56
Objective:
To investigate the delay in identification, healthcare-seeking, and definitive diagnosis of tuberculosis among students in Urumqi City from 2010 to 2019, and to identify the influencing factors, so as to provide insights into tuberculosis control among students.
Methods:
The demographic and diagnosis data of tuberculosis patients in Urumqi City from 2010 to 2019 were captured from the Tuberculosis Information Management System of Chinese Disease Control and Prevention Information System. The delay in identification, healthcare-seeking and definitive diagnosis of tuberculosis was analyzed among students, and the factors affecting the delay in identification, healthcare-seeking and definitive diagnosis of tuberculosis were identified using a multivariable logistic regression model.
Results:
A total of 996 tuberculosis cases were identified among students in Urumqi City from 2010 to 2019. There were 702 students with delay in identification of tuberculosis (70.48%), 500 students with delay in healthcare-seeking (55.22%) and 534 students with delay in definitive diagnosis (53.61%). Multivariable logistic regression analysis identified active identification (OR=0.116, 95%CI: 0.032-0.420) as a factor affecting delay in identification of tuberculosis, women (OR=1.424, 95%CI: 1.104-1.836), non-local household registration (OR=1.311, 95%CI: 1.016-1.694) and active identification (OR=0.232, 95%CI: 0.064-0.848) as factors affecting delay in healthcare-seeking, and active identification (OR=0.143, 95%CI: 0.032-0.644) as a factor affecting delay in definitive diagnosis of tuberculosis among students.
Conclusions
There is a high proportion of delay in identification, healthcare-seeking and definitive diagnosis of tuberculosis among students in Urumqi City from 2010 to 2019, and female and non-locally household-registered students were at a high risk of delay in healthcare-seeking for tuberculosis. Active detection and screening of tuberculosis should be reinforced.
8.MicroRNA-130a Increases and Predicts Cardiotoxicity during Adjuvant Chemotherapy in Human Epidermal Growth Factor Receptor-2-Positive Breast Cancer
Qiang FENG ; Yanbin REN ; Aijun HOU ; Jing GUO ; Zhezhe MAO ; Shaojun LIU ; Boya WANG ; Zhichao BAI ; Xiaoying HOU
Journal of Breast Cancer 2021;24(2):153-163
Purpose:
This study aimed to investigate the changes in microRNA-130a (miR-130a) and its correlation with cardiotoxicity during epirubicin/cyclophosphamide followed by docetaxel plus trastuzumab (EC-D+T) adjuvant chemotherapy in human epidermal growth factor receptor-2-positive (HER2+) breast cancer patients.
Methods:
A total of 72 HER2+ breast cancer patients who underwent resection and were scheduled to receive EC-D+T adjuvant therapy were consecutively enrolled. The expression of miR-130a and cardiotoxicity (defined as any of the following situations: 1) absolute decline of left ventricular ejection fraction (LVEF) ≥ 10% and LVEF < 53%; 2) heart failure; 3) acute coronary artery syndromes; and 4) fatal arrhythmia) were assessed every 3 months throughout the 15-month EC-D+T treatment.
Results:
The accumulating cardiotoxicity rate was 12 (16.7%), of which the incidence of heart failure, acute coronary syndrome, life-threatening arrhythmias, ΔLVEF ≥ 10%, and LVEF < 53% was 0 (0.0%), 1 (1.4%), 0 (0.0%), and 12 (16.7%), respectively. Baseline miR-130a expression was negatively correlated with LVEF (%) and positively correlated with cardiac troponin I. The expression of miR-130a gradually increased in both cardiotoxicity and noncardiotoxicity patients during EC-D+T treatment, while the increment of miR-130a was more obvious in cardiotoxicity patients compared with non-cardiotoxicity patients. Further logistic regression and receiver operating characteristic curve analysis indicated that miR-130a was an independent predictive factor for increased cardiotoxicity risk.
Conclusion
MiR-130a increases constantly and predicts high cardiotoxicity risk during ECD+T adjuvant chemotherapy in HER2+ breast cancer patients.
9.MicroRNA-130a Increases and Predicts Cardiotoxicity during Adjuvant Chemotherapy in Human Epidermal Growth Factor Receptor-2-Positive Breast Cancer
Qiang FENG ; Yanbin REN ; Aijun HOU ; Jing GUO ; Zhezhe MAO ; Shaojun LIU ; Boya WANG ; Zhichao BAI ; Xiaoying HOU
Journal of Breast Cancer 2021;24(2):153-163
Purpose:
This study aimed to investigate the changes in microRNA-130a (miR-130a) and its correlation with cardiotoxicity during epirubicin/cyclophosphamide followed by docetaxel plus trastuzumab (EC-D+T) adjuvant chemotherapy in human epidermal growth factor receptor-2-positive (HER2+) breast cancer patients.
Methods:
A total of 72 HER2+ breast cancer patients who underwent resection and were scheduled to receive EC-D+T adjuvant therapy were consecutively enrolled. The expression of miR-130a and cardiotoxicity (defined as any of the following situations: 1) absolute decline of left ventricular ejection fraction (LVEF) ≥ 10% and LVEF < 53%; 2) heart failure; 3) acute coronary artery syndromes; and 4) fatal arrhythmia) were assessed every 3 months throughout the 15-month EC-D+T treatment.
Results:
The accumulating cardiotoxicity rate was 12 (16.7%), of which the incidence of heart failure, acute coronary syndrome, life-threatening arrhythmias, ΔLVEF ≥ 10%, and LVEF < 53% was 0 (0.0%), 1 (1.4%), 0 (0.0%), and 12 (16.7%), respectively. Baseline miR-130a expression was negatively correlated with LVEF (%) and positively correlated with cardiac troponin I. The expression of miR-130a gradually increased in both cardiotoxicity and noncardiotoxicity patients during EC-D+T treatment, while the increment of miR-130a was more obvious in cardiotoxicity patients compared with non-cardiotoxicity patients. Further logistic regression and receiver operating characteristic curve analysis indicated that miR-130a was an independent predictive factor for increased cardiotoxicity risk.
Conclusion
MiR-130a increases constantly and predicts high cardiotoxicity risk during ECD+T adjuvant chemotherapy in HER2+ breast cancer patients.
10.The efficacy and complications of minimally invasive vs. the traditional open transforaminal lumbar interbody fusion for the treatment of lumbar spondylolisthesis
Aimin WU ; Zhichao HU ; Zhenhua FENG ; Xiaobing LI ; Hui XU ; Shen WANG ; Qishan HUANG ; Fangmin MAO ; Yan LIN ; Xiangyang WANG ; Wenfei NI
Chinese Journal of Orthopaedics 2018;38(20):1230-1239
Objective To investigate the clinical efficacy and complications of minimally invasive transforaminal lumbar-interbody fusion (TLIF) in the treatment of lumbar spondylolisthesis. Methods Total 142 patients with single level spondylolis-thesis who treated by TLIF from 2010.01 to 2015.06 were included in this study, with 68 cases in minimally invasive TLIF (MIS-TLIF) group and 74 cases in traditional open TLIF group. The general information (age, gender, isthmic or degenerative type, per-centage of slip degree, levels), operative time, blood loss, length of postoperative hospital stay, Visual Analogue Scale (VAS) of low-back pain and leg pain, and Oswestry Disability Index (ODI) were recorded and collected. The posterior height of the interverte-bralpace and segmental lordosis, reduction of spondylolisthesis and cross-sectional area of spinal canal were measured. Results There was no statistically significant difference between the two groups in age, gender ratio, percentage of slip degree, and sur-gicallevels distribution. Total of 66 cases in MIS-TLIF group and 71 cases in Open TLIF group finished 2 years follow up, and 25 cases in MIS-TLIF group and 31 cases in Open TLIF group finished 5 years follow up. The blood loss of the MIS-TLIF group was 164.7±51.7 ml, significantly lower than the open TLIF group of 239±69.3 ml(P<0.001). The length of postoperative hospital stay was 5.9 ± 1.5 days in MIS-TLIF group, significantly shorter than the open TLIF group of 7.3 ± 3.1 days(P<0.001). The operative time of MIS-TLIF and Open TLIF was 146.3±21.9 mins, 152.0±20.4 mins, respectively, and no significant differ-ence was found between them. The VAS ofback pain, leg pain, ODI in MIS-TLIF group was 1.76±1.16, 1.91±1.36 and 23.5± 7.3 at 2 years follow up, and in Open TLIF was 1.73±1.10, 1.83±1.36 and 23.8±6.7, respectively, all of them were significant-ly different to pre-operation, however, no significant difference was found between two groups. The VAS of back pain, leg pain, ODI in MIS-TLIF group was 1.73±1.21, 1.93±1.48, and 25.4±6.8 at 5years follow up, and in Open TLIF was 1.85±1.02, 1.85± 1.33 and 26.1 ± 6.5, respectively, no significant difference between twogroups. The posterior height of the intervertebral space and segmental lordosis of MIS-TLIF was 9.52±1.67 mm and 12.11°±3.44° at 2 years follow up, while the open TLIF was 9.88± 1.54 mm and 12.98 ± 3.83° , all of them were significantly different to pre-operation,however, no significant difference between two groups. The posterior height of the intervertebral space and segmental lordosis of MIS-TLIF was 9.37 ± 1.46 mm and 11.55° ± 2.77° , while the open TLIF was 9.66 ± 1.68 mm and 12.59° ± 4.23° , no significant difference between two groups. The percentage of slip degree was reduced to 5.2%±4.6% in MIS-TLIF and 5.6%±4.3% in open TLIF, the cross-sectional area of spinal canal was enlarged to 139.7±19.5 mm2 and 141.7±20.7 mm2, no significant difference between two groups either. Con-clusion MIS-TLIF has less blood loss, shorter postoperative hospital stay than open TLIF, and similar clinical pain and function-al outcomes. MIS-TLIF is suggested to be a safe and effective choice in the treatment of lower grade lumbar spondylolisthesis (Grade II or less).


Result Analysis
Print
Save
E-mail