1.Roles of Keap1-Nrf2 and NF-κB signaling pathways in lipopolysaccharide-induced brain injury
Yichen LIU ; Dongfei YAN ; Zhixin LI ; Yingchun MAO ; Zhihui LI ; Guofu DONG ; Changzhen WANG
Military Medical Sciences 2024;48(7):481-486
Objective To investigate the role of Kelch-like-epichlorohydrin-associated protein 1/nuclear factor erythroid-derived factor-2-related factor 2(Keap1-Nrf2)and nuclear factor-κB(NF-κB)signaling pathways in sepsis-associated encephalopathy(SAE).Methods Male C57BL/6J mice of SPF were randomly divided into four groups(n=10):the control group and LPS 6 h,24 h and 48 h groups.The behavioral changes of the mice were assessed based on their general conditions and open field test(OFT).ELISA was used to measure the levels of pro-inflammatory cytokines in mouse serum,and the antioxidant capacity assay kit to examine antioxidant activity in brain tissues of mice.Real-time quantitative PCR(qPCR)was adopted to detect the mRNA levels of toll-like receptor4(Tlr4),NF-κB,Keap1 and Nrf2 in the hippocampus,and to determine protein expressions of NF-κB a Nrf2、Keap1 and Tlr4 with Western blotting.Results Compared to the control group,the serum concentrations of interleukin 6(IL-6)in lipopolysaccharide(LPS)groups increased at 6 h,and reached the peak at 24 h and 48 h(P<0.01).The levels of serum interleukin 18(IL-18)in the LPS groups increased significantly at 6 h and 24 h(P<0.01)but there was no statistically significant difference compared with the 48h group.The results indicated the activity of superoxide dismutase(SOD)and glutathione(GSH)in brain tissues in LPS groups increased(P<0.01).OFT results showed the time spent in the center of the open field,the distance covered around the center,and total distance covered by mice in LPS groups were significantly reduced(P<0.01),except for the time spent in the center of the open field in the LPS 24 h group.The mRNA expressions of Tlr4 and(LPS 6 h,48 h)NF-κB in the hippocampus tissue of mice in LPS groups were elevated(P<0.05),so were the mRNA expressions of Keap1 and Nrf2 in LPS 6 h group.Additionally,the protein expressions of NF-κB,Keap1 and Tlr4 increased in LPS groups,so did the protein expression of Nrf2 in LPS 24 h and 48 h groups(P<0.05).Conclusion Keap1-Nrf2 and NF-κB signaling pathways may play a certain role in SAE.
2.Influencing factors for unplanned re-hospitalization in adult renal transplant recipients after surgery
Weiwei CAO ; Minghuan ZHONG ; Zhou SUN ; Guofu LIANG ; Fu YAN ; Chao LIU ; Li MA ; Kejing ZHU ; Yanyan XU ; Bei DING ; Yulin NIU
Journal of Clinical Medicine in Practice 2024;28(18):117-122
Objective To investigate the influencing factors of unplanned rehospitalization within one year after surgery among adult renal transplant recipients. Methods The clinical data of 299 recipients who underwent renal transplant surgery in the Department of Organ Transplantation of the Affiliated Hospital of Guizhou Medical University from January 2020 to December 2022 were retrospectively analyzed. The recipients were divided into unplanned rehospitalization group and non-rehospitalization group based on whether they experienced unplanned rehospitalization within one year after surgery. Univariate analysis and binary Logistic regression analysis were performed to explore the influencing factors of unplanned rehospitalization within one year after renal transplantation. Results Among the 299 recipients, 102 experienced unplanned rehospitalization, with an incidence rate of 34.11%. Univariate analysis revealedstatistically significant differences were noted between the two groups in terms of gender, occupational status, preoperative underlying disease, rejection reactions, nosocomial infections, immunosuppressive medication regimens, serum creatinine, cystatin C, serum phosphorus, serum potassium, and initial hospitalization duration (
3.The effects of leptin downregulation by RNA interference on the proliferation ability transforming growth factor-β 1 and type Ⅰ collagen expression levels of pathological scar fibroblasts
Xiaojuan CHEN ; Guofu ZHOU ; Yan CAI ; Xiaoyan XIANG
Chinese Journal of Plastic Surgery 2022;38(2):215-221
Objective:To investigate the effects of RNA interference on the expression level of leptin and its effect on proliferation, TGF-β 1 and collagen type Ⅰ production in human pathological scar fibroblasts in vitro. Methods:Pathological scar tissues (hypertrophic scar, keloid) were collected from the Department of Burn Plastic Surgery in Affiliated Hospital of North Sichuan Medical College. After primary cell culture and cell passage, passage 3 cells were selected for experimental study. The cells were divided into two groups: the experimental group which was transfected with leptin siRNA, and the negative control group which was transfected with empty vector. Examinations were carried out 48 hours after transfection. Cell proliferation was determined by CCK-8. The transcription levels of leptin, TGF-β 1 and type Ⅰ collagen genes were detected by polymerase chain reaction (PCR). The protein expression levels of leptin, TGF-β 1 and type Ⅰ collagen were determined by immunofluorescence and Western blotting. Student′s t-test was used for comparison between groups, and P<0.05 was considered statistically significant. Results:Five hypertrophic scars and five keloids were included. Compared with the negative control groups, the proliferation ability ( A450) of leptin-SiRNA transfected fibroblasts were not significantly different ( P>0.05). The relative mRNA expression levels of leptin, TGF-β 1 and type Ⅰ collagen in hypertrophic scars and keloids were significantly decreased in the siRNA transfection groups compared with the negative control groups ( P<0.05). Immunofluorescence results showed that the expression levels of leptin, TGF-β 1 and type I collagen in keloids were higher than those in hypertrophic scars, and that siRNA induced leptin downregulation significantly reduced the levels of leptin, TGF-β 1 and type I collagen in both hypertrophic scars and keloids. Western blotting showed that the protein levels of leptin, TGF-β 1 and type Ⅰ collagen were significantly decreased in hypertrophic scar and keloid fibroblasts after leptin siRNA interference ( P<0.05). Conclusions:Downregulation of leptin gene expression by RNA interference inhibited TGF-β 1 and typeⅠ collagen expression, which could be used in treating pathological scar.
4.The effects of leptin downregulation by RNA interference on the proliferation ability transforming growth factor-β 1 and type Ⅰ collagen expression levels of pathological scar fibroblasts
Xiaojuan CHEN ; Guofu ZHOU ; Yan CAI ; Xiaoyan XIANG
Chinese Journal of Plastic Surgery 2022;38(2):215-221
Objective:To investigate the effects of RNA interference on the expression level of leptin and its effect on proliferation, TGF-β 1 and collagen type Ⅰ production in human pathological scar fibroblasts in vitro. Methods:Pathological scar tissues (hypertrophic scar, keloid) were collected from the Department of Burn Plastic Surgery in Affiliated Hospital of North Sichuan Medical College. After primary cell culture and cell passage, passage 3 cells were selected for experimental study. The cells were divided into two groups: the experimental group which was transfected with leptin siRNA, and the negative control group which was transfected with empty vector. Examinations were carried out 48 hours after transfection. Cell proliferation was determined by CCK-8. The transcription levels of leptin, TGF-β 1 and type Ⅰ collagen genes were detected by polymerase chain reaction (PCR). The protein expression levels of leptin, TGF-β 1 and type Ⅰ collagen were determined by immunofluorescence and Western blotting. Student′s t-test was used for comparison between groups, and P<0.05 was considered statistically significant. Results:Five hypertrophic scars and five keloids were included. Compared with the negative control groups, the proliferation ability ( A450) of leptin-SiRNA transfected fibroblasts were not significantly different ( P>0.05). The relative mRNA expression levels of leptin, TGF-β 1 and type Ⅰ collagen in hypertrophic scars and keloids were significantly decreased in the siRNA transfection groups compared with the negative control groups ( P<0.05). Immunofluorescence results showed that the expression levels of leptin, TGF-β 1 and type I collagen in keloids were higher than those in hypertrophic scars, and that siRNA induced leptin downregulation significantly reduced the levels of leptin, TGF-β 1 and type I collagen in both hypertrophic scars and keloids. Western blotting showed that the protein levels of leptin, TGF-β 1 and type Ⅰ collagen were significantly decreased in hypertrophic scar and keloid fibroblasts after leptin siRNA interference ( P<0.05). Conclusions:Downregulation of leptin gene expression by RNA interference inhibited TGF-β 1 and typeⅠ collagen expression, which could be used in treating pathological scar.
5.Low molecular weight heparin through TLR4/MyD88/NF-κB pathway protects PC12 cells against oxygen glucose deprivation induced inflammation
Guofu LI ; Yan CHEN ; Wangbin DENG ; Yongkang ZHANG ; Yanping GUO ; Da MA ; Zhuo CHEN ; Yuan LIANG
Journal of Chinese Physician 2021;23(6):848-852,858
Objective:To investigate the effect of low molecular weight heparin (LMWH) on the inflammatory response of PC12 cells induced by oxygen glucose deprivation (OGD) and its related mechanism.Methods:The PC12 cells were cultured in vitro were randomly divided into sham(control) group, OGD group, LMWH group and blocking agent group. The latter group was divided into six groups: Eritoran+ OGD group, LMWH+ Eritoran+ OGD group, ST2825+ OGD group, LMWH+ ST2825+ OGD group, pyrrolidinedithiocarbamate (PDTC)+ OGD group and LMWH+ PDTC+ OGD group. OGD cell model was established. Cell counting kit-8 (CCK-8) assay was used to detect cell activity. The expressions of toll-like receptor 4 (TLR4), MyD88 and nuclear factor κB (NF-κB) mRNA and protein were detected by real time polymerase chain reaction (qRT-PCR) and Western blot. The concentration of interleukin (IL)-1β, IL-6, tumor necrosis factor-α(TNF-α) and S100β were determined by enzyme linked immunosorbent assay (ELISA). Results:The cell activity of OGD group was significantly lower than that of control group on the first, second, third day ( P<0.05). Compared with OGD group, the activity of LMWH group was increased on the second, third day ( P<0.05), but lower than that of control group ( P<0.01). The mRNA expression of TLR4, MyD88 and NF-κB was significantly increased in OGD group compared with the control group ( F=144.9, F=710.5, 79.51, P<0.01). Compared with OGD group, the mRNA expression of TLR4, MyD88 and NF-κB were significantly decreased after treatment with LMWH ( P<0.01), and the specific inhibitor of TLR4, MyD88 and NF-κB enhanced the anti-inflammatory effect of LMWH. The protein expression of this pathway was consistent with that of the gene. The concentration of IL-1β, IL-6, TNF-α and S100β in OGD group was significantly higher than control group ( P<0.05). After treatment with LMWH, the concentrations of inflammatory factors and S100β were significantly decreased compared with OGD group ( P<0.01). When hinder TLR4, MyD88 and NF-κB respectively by Eritoran, ST2825 and PDTC, the concentrations of inflammatory factors and S100β were significantly decreased, but it was still higher than control group ( P<0.05). Conclusions:OGD can cause pathological damage of PC12 cells, including high expression level of S100β and aggravation of inflammatory reaction. LMWH can improve cell activity, down-regulate inflammatory reaction degree and protect the cells. Using inhibitors of TLR4/MyD88/NF-κB pathway to inhibit the corresponding target, the up-regulation of inflammatory factors by OGD can be inhibited in varying degrees. These suggested that LMWH may regulate inflammatory reaction of PC12 cells induced by OGD through TLR4/MyD88/NF-κB pathway.
6.Clinical application of new type individualized fixation device for radiotherapy of rectal cancer
Guofu CHEN ; Guiming YAN ; Feiyan ZHANG
Chinese Journal of Primary Medicine and Pharmacy 2019;26(1):2-5
Objective To explore the clinical effect of new type rectal cancer radiotherapy individualized fixation device in the radiotherapy of rectal cancer. Methods From June 2015 to December 2016,60 patients with rectal cancer who accepted the external irradiation in Zhejiang Tumor Hospital were divided into two groups by random number table method. A group(31 cases) received new type rectal cancer radiotherapy position fixation devices with thermoplastic film. B group(29 cases) received simple foam pad with thermoplastic film. Before each treatment,Cone beam CT(CBCT) scan was conducted. The applied CBCT image and the planned reconstruction image were compared in the direction of X(left and right),Y(upper and lower)and Z(front and rear) axis. The setup error was recorded, and the correlation between the two groups was analyzed. Results The average setup error of patients in A group in X (left and right),Y(upper and lower),Z(front and rear) axis were (1. 61 ± 0. 18)mm,(1. 82 ± 0. 13)mm,(1. 91 ± 0. 11)mm,respectively. The average setup error of patients in B group in X(left and right),Y(upper and lower),Z (front and rear) axis were (2. 22 ± 0. 13)mm,(2. 43 ± 0. 14)mm,(2. 36 ± 0. 13)mm,respectively. There were statistically significant differences between the two groups(t=14. 958,17. 501,11. 283,all P<0. 001). Conclusion The new type of rectal cancer radiotherapy position fixing device is more comfortable than the simple foam pad,and the setting error is smaller than the simple foam pad.
7.MRI measurement for normal fetal brain biometric parameters in second and third trimesters
Xiaoliang MA ; Guofu ZHANG ; Yan ZHUANG ; Xuefen LIU
Chinese Journal of Perinatal Medicine 2018;21(11):724-730
Objective To analyze the correlations of commonly used biometric parameters of normal fetal brain with fetal gender and gestational age (GA) and to evaluate the feasibility of establishing MRI reference standards for normal fetal brain biometry in second and third trimesters. Methods MRI measurements of 263 fetuses without central nervous system abnormalities were retrospectively collected from the Obstetrics and Gynecology Hospital of Fudan University from June 2012 to April 2017. MRI measurements of fetal biparietal diameter (BPD), occipital-frontal diameter (OFD), head circumference cross-sectional area (HCS), transverse cerebellar diameter (TCD), anterior-posterior diameter of the vermis (APDV), vermian height (VH) and vermian cross-sectional area (VS) were obtained in every standard plane. Head circumference (HC) and (BPD+OFD)/2 were calculated. Spearman correlation analysis, independent samples t-test, Pearson correlation analysis and linear regression analysis were used for statistic analysis. Results Fetal BPD, OFD and HCS were statistically correlated with gender in second and third trimesters (r= - 0.155, - 0.149 and- 0.159; P=0.032, 0.038 and 0.027). The mean values of BPD, OFD and HCS of male fetuses were greater than those of female ones [(7.25±1.22) vs (6.87±1.28) cm, (8.59±1.38) vs (8.18±1.41) cm, (53.62±16.82) vs (48.48±16.84) cm2; t=2.101, 2.011 and 2.102; all P<0.05] and the mean differences between them were 0.38 cm, 0.41 cm and 5.14 cm2, respectively. BPD, OFD, HCS, HC, (BPD+OFD)/2, TCD, APDV, VH and VS were positively correlated with gestational age (r=0.950, 0.947, 0.962, 0.957, 0.957, 0.976, 0.931, 0.943 and 0.960, respectively; all P<0.001). Among them, TCD (r=0.976) was the closest relating factor, followed by HCS (r=0.962) and VS (r=0.960). The linear regression equations were as follows: BPD (cm)=0.276×GA-0.453, R2=0.903; OFD (cm)=0.308×GA+0.010, R2=0.896; (BPD+OFD)/2 (cm)=0.292×GA - 0.222, R2=0.916;HC (cm)=0.473×GA - 0.359, R2=0.916; HCS (cm2)=3.795×GA - 52.232, R2=0.926; TCD (cm)=0.180× GA-1.858, R2=0.952; APDV (cm)=0.047×GA-0.353, R2=0.867; VH (cm)=0.071×GA-0.592, R2=0.890;VS (cm2)=0.143×GA-2.396, R2=0.919, all P<0.001. Conclusions Some fetal brain biometric parameters are correlated with fetal gender. It is essential and conducive to establish different reference values for male and female fetuses to achieve accurate prenatal evaluation and diagnosis. Fetal BPD, OFD, HCS, HC, (BPD+OFD)/2, TCD, APDV, VH and VS have strong linear relationship with gestational age in second and third trimester and can be served as good indicators for evaluating fetal brain growth and development. MRI quantitative measurement of fetal brain biometric parameters can provide more reliable reference data for prenatal evaluation of fetal brain growth and development. It is feasible to establish MRI reference standards for fetal brain biometry.
8.Application of interventional embolization of fallopian tube in the prevention of tubal stump pregnancy
Tianping WANG ; Zhiping YAN ; Guofu ZHANG
Journal of Interventional Radiology 2018;27(2):147-150
Objective To investigate the clinical characteristics of tubal stump pregnancy, and to assess the value of interventional embolization in preventing tubal stump pregnancy. Methods Among the patients who were planned to receive preoperative treatment of hydrosalpinx before the performance of in vitro fertilization-embryo transfer (IVF-ET) at authors' hospital, 35 patients had stump of fallopian tube. Of the35 patients, previous surgery of unilateral fallopian tube was present in 28 and previous surgery of bilateral fallopian tubes in 7. The length of tubal stump ranged from 10mm to 45mm, with a mean of 25mm. Interventional embolization with micro-coils was carried out in all patients, after which IVF-ET was performed. The pregnancy rate and ectopic pregnancy rate were calculated. Results Embolization of both fallopian tubes was successfully accomplished in all 35 patients. The IVF-ET pregnancy rate was 48.5% (17/35), and no tubal stump pregnancy occurred. Conclusion The use of interventional embolization to occlude tubal stump can effectively avoid the occurrence of tubal stump pregnancy.
9.Survey on cognitive status quo of nursing staffs on patients safety culture in second-class and tertiary hospitals of Nanchang City
Jingjing ZHANG ; Xia QIAN ; Yuan CHEN ; Guofu HUANG ; Yan LIU ; Ping ZHANG ; Yingshan SHI ; Fengbao XIONG
Chongqing Medicine 2017;46(12):1659-1662,1665
Objective To investigate the current situation of nursing staffs in second-class and tertiary hospitals of Nanchang City and influencing factors in order to provide a basis for the management and research of patients safety.Methods The convenience cluster sampling method was adopted to extract the whole nursing staffs from 7 hospitals(4 second-class hospitals and 3 tertiary hospitals)in Nanchang City as the research subjects.Then the multiple regression method was adopted to analyze the influencing factors of patients safety culture in the second-class and tertiary hospitals of Nanchang City.Results The advantage fields of cognition of the nursing staffs in the second-class hospitals on the patients safety culture were the intra-department teamwork,expectation actions of managers for promoting safety,organizational learning and continuous improvement,feedback and communication of mistakes,while the fields needing to be improved were the communication openness,personnel allocation,non-penalty reactions on mistakes;the advantage fields in the tertiary hospitals were same to those in the second-class hospitals,and the fields needing to be improved included the inter-department cooperation,personnel allocation,non-penalty reactions on mistakes.Conclusion The cognition of nursing staffs in 7 hospitals of Nanchang City on the patients safety culture is in a higher level,but insufficiency still exists,partial fields need to be continuously improved.
10.Analysis on relationship between serum anti-desmoglein antibodies and disease in patients with pemphigus
Dan KE ; Shuqing DENG ; Kaiyan CAI ; Shudan HE ; Xi WANG ; Honglin LI ; Guofu YAN
Chongqing Medicine 2016;45(18):2498-2499,2503
Objective To investigate the application value of ELISA for detecting the serum anti desmoglein (Dsg) 1 and Dsg 3 in the diagnosis and treatment of pemphigus .Methods Forty‐seven patients with pemphigus in our hospital from January to De‐cember 2014 were selected as the observation group and contemporaneous 52 patients with excluding pemphigus were selected as the control group .The Dsg antibodies were detected by using indirect immunofluorescence method and Dsg 1 and Dsg3 were deter‐mined by ELISA ;their correlation with pemphigus characteristics was analyzed .Results The sensitivity and specificity of ELISA for detecting anti‐Dsg antibodies were 95 .74% and 92 .31% respectively ,while which of IIF were 93 .62% and 86 .54% respective‐ly ,showing no statistically significant difference between the two test methods (P>0 .05) .In 30 cases of pemphigus vulgaris ,16 ca‐ses (16/30) were positive Dsg1 and Dsg 3 ,8 cases of pemphigus erythematosus and 5 cases pemphigus foliaceus were positive Dsg1 only ,and 2 cases of pemphigus vegetans were both positive Dsgl and Dsg3 .The Dsgl and Dsg3 titers of pemphigus vulgaris and pemphigus vegetans were 130 .85 ± 86 and 112 .30 ± 85 .05 ,respectively ,and the disease activity score was (5 .10 ± 1 .86) points ,the correlation coefficient(r)=0 .476(P=0 .008) ,r=0 .816(P=0 .001) ,respectively .The Dsgl titer of pemphigus erythematosus and pemphigus foliaceus were 142 .59 ± 78 .52 ,and the disease activity score was (2 .77 ± 0 .92) points(r=0 .800 ,P=0 .001) .Conclu‐sion ELISA for detecting Dsg1 and Dsg3 has high sensitivity and specificity ,and is conducive to the diagnosis of pemphigus and e‐valuation of disease severity .


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