1.miR-520d over-expression reverses chemotherapy resistance of TNBC via inhibiting autophagy protein Beclin1
LIU Huan ; LI Hongchang ; CHEN Yafeng ; XU Ke ; LI Jie ; FENG Dianxu
Chinese Journal of Cancer Biotherapy 2019;26(10):1062-1067
Objective: To investigate the role and molecular mechanism of miR-520d in reversing the chemoresistance of triple negative breast cancer (TNBC) by regulating autophagy. Methods: Docetaxel (Doc) resistant cell lines MDA-MB-231/Doc and MDA-MB468/Doc were constructed by using human TNBC cell lines MDA-MB-231 and MDA-MB-468 as parental cells, and the cells were divided into blank group (parental cells), control group (drug-resistant group), and miR-520d over-expression group. The expression levels of miR-520d in cells of the blank and drug-resistant groups were detected by qPCR. The Doc-sensitivity of resistant cells over-expressing miR-520d was detected by MTT assay.After MDC staining, the generation of autophagosome in cells was observed under fluorescence microscopy; the number of miR-520d over-expressed resistant cells with positive LC3 expression was observed under confocal microscopy. The luciferase reporter gene assay was used to verify the targeting relationship between miR-520d and Beclin1. The effect of miR-520d mimics on the expression of autophagy-associated protein Beclin1, and LC3Ⅰ, LC3Ⅱ in cells was detected by WB assay. Results: The results of qPCR showed that the expression of miR-520d in the drug-resistant TNBC cells was significantly lower than that of normal cells (P<0.01). In drug-resistant cells over-expressing miR-520d, the Doc-sensitivity was significantly improved, while the autophagy activity was significantly reduced (all P<0.01).At the same time, luciferase experiments demonstrated that Beclin1 was a possible target molecule of miR-520d (P<0.05). WB results showed that the combination of docetaxel and miR-520d mimics reduced the LC3-II/I ratio and the expression of autophagy protein Beclin1 in drug-resistant TNBC cells (all P<0.05). Conclusion: The regulation of miR-520d levels may alter the expression of autophagy protein Beclin1, thereby reversing Doc chemotherapy resistance in TNBC cells.
2.Effects of Huanglian Ointment promoting Angiogenesis by PI3K/AKT/eNos pathway in Full-thickness Skin Defect Mice
Xiaofen ZHANG ; Jing SONG ; Hongchang LI ; Yafeng CHEN ; Dianxu FENG
World Science and Technology-Modernization of Traditional Chinese Medicine 2018;20(4):527-533
Objective: to observe the effects of Huanglian ointment promote wound healing and angiogenesis by the AKT/VEGF/eNos pathway in full-thickness skin defect mice. Methods: 7.5 mm diameter full-thickness skin excision modelwas made in the back of the 45 male C57 BL/6 J mice respectively. That were subsequently randomly placed into 3 groupswith Random number table method; i.e., vehicle, Huanglian ointment groups and the control group. In the Huanglianointment group, topical Huanglian ointment was applied to the wound, in the vehicle group were treated with vehicleointment, and in the control group were treated with nothing. Changes in the size of their wounds was monitored by takingpictures with a digital camera on days 0, 3, 7, 10, and 14 after wound creation. The mice were sacrificed on the 3, 7, and14 days after wound creation, and the tissue samples of the wounds were obtained for m RNA level of b FGF and PDGF、CD-31 cells and expression of AKT、VEGF-A、eNos were measured too. Results: Comparison of the sizes of the woundsamong the groups showed that there was no significant difference on the 0, 3 and 7 days, the most significant decreaseswere found in experimental Huanglian ointment group on the day10 (Huanglian ointment versus vehicle: (76±7) % VS (48±9) %, huanglian ointment versus control: (76±7) % VS (46±7) %, P<0.01), and day14: (Huanglian ointment versus vehicle: (93±5) % VS (68 ±11) %, huanglian ointment versus control: (93 ±5) % VS (64±9) %, P<0.01) . The percentage of CD-31 cells on the Huanglian ointment group were significantly higher than that of the vehicle and control groups on the 3、7 days, (Huanglian ointment versus vehicle: day3: (16.3±3.2) % VS (12.5±4.6) %, P<0.05;day7: (33.6±5.0) % VS (19.2±4.0) %, P<0.01; (Huanglian ointment versus control: day3: (16.3±3.2) % VS (8.4±2.4) %, P<0.05;day7: (33.6±5.0) % VS (17.8±6.0) %, P< 0.01. The m RAN of b FGF on the Huanglian ointment group were significantly higher than the vehicleand control groups on the 3 and 7 days, (day3: Huanglian ointment versus vehicle: (1.75±0.22) VS (0.96±0.13), day7: (2.98±0.35) VS (1.53±0.24), P<0.01) ); (day3: Huanglian ointment versus control (1.75±0.22) VS (0.78±0.24), and day7: (2.98 ± 0.35) VS (1.64 ± 0.31), P<0.01) . But on 14 day, the vehicle and control groups were significantly higher thanHuanglian ointment group, Huanglian ointment versus vehicle (1.43±0.42) VS (1.88±0.38), Huanglian ointment versuscontrol (1.43±0.42) VS (2.03±0.21), P< 0.05. The m RAN of PDGF on the Huanglian ointment group were significantlyhigher than the vehicle and control groups on the 3、7 days, (day3: Huanglian ointment versus vehicle (1.04±0.28) VS (0.56±0.15), Huanglian ointment versus control (1.04±0.28) VS (0.67±0.20) (P<0.01); day7: Huanglian ointment versusvehicle (1.82±0.25) VS (1.38±0.21), Huanglian ointment versus control (1.82±0.25) VS (1.45±0.26) (P<0.05) . On the 7 day, the protein of P-AKTS308 and P-AKTS437 in wound tissue on the Huanglian ointment group were significantlyhigher than the vehicle and control groups, (P-AKTS308: Huanglian ointment versus vehicle: (0.45±0.04) VS (0.23±0.06), Huanglian ointment versus control: (0.45 ± 0.04) VS (0.19 ± 0.08), (P<0.05); (P-AKTS437: Huanglian ointmentversus vehicle: (0.27±0.03) VS (0.16±0.04); Huanglian ointment versus control: (0.27±0.03) VS (0.20±0.05), (P<0.01) .the protein of VEGF-A and e NOS on the Huanglian ointment group were significantly higher than the vehicle and controlgroups too, Huanglian ointment versus vehicle: (VEGF-A: (0.18±0.02) VS (0.26±0.04), P<0.01, e NOS: (0.12±0.05) VS (0.14±0.07, P<0.01) ); Huanglian ointment versus control: (VEGF-A: (0.18±0.02) VS (0.13±0.06), P<0.01, e NOS: (0.12±0.05) VS ((0.17±0.03), (P<0.01) ) . Conclusions: Huanglian ointment promotes wound healing and enhance b FGF, PDGFand VEGF-A content by increasing the angiogenesis with the AKT/VEGF/eNos pathway.
3.Shengjiyuhong ointment inhibits hypertrophic scar formation
Guifang SUN ; Xiaofen ZHANG ; Hongchang LI ; Liyun PAN ; Yafeng CHEN ; Ke XU ; Dianxu FENG
Chinese Journal of Tissue Engineering Research 2016;20(33):4890-4898
BACKGROUND:Shengjiyuhong ointment has been reported to inhibit hypertrophic scarring. OBJECTIVE:To verify the effects of Shengjiyuhong ointment on hypertrophic scarring of in a rabbit ear model. METHODS:Each ear of thirty-six Japanese rabbits was used to make four 1-cm-diameter circular ful-thickness skin wounds with the entire perichondrium removed. Final y, 288 wounds were made and randomly divided into 6 groups:model, negative control (no drugs were administered), low-, moderate-, high-crude herbal dose drugs (Shengjiyuhong ointment was administered topical y at concentrations of 8.39%, 25.18%, and 75.54%), and positive control (recombinant bovine basic fibroblast growth factor was administered topical y). Shengjiyuhong ointment was administered twice daily til wound healing. The wounds were evaluated by the Vancouver scar scale (VSS). Scar elevation index (SEI) of scar specimens was calculated under a microscope at 40× magnification. mRNA expression levels of type I and III col agen, connective tissue growth factor, fibronectin, andα-smooth muscle actin (α-SMA) were determined by fluorescent quantitative PCR. Protein expression levels of type I and III col agen andα-SMA were detected by western blot assay.α-SMA immunoreactivity was determined by immunofluorescent staining. RESULTS AND CONCLUSION:VSS scores and SEI were significantly increased in each group at 30 days (P<0.05). VSS scores and SEI were significantly decreased in the moderate-and high-crude herbal dose drug groups and positive control groups compared with the model, negative control, and low-crude herbal dose drug groups (P<0.05 or P<0.01). mRNA expression levels of type I and III col agen, connective tissue growth factor and fibronectin, and protein expression levels of type I and III col agen andα-SMA were significantly inhibited after moderate-crude herbal dose Shengjiyuhong ointment and positive drug treatment (P<0.01). These findings suggest that Shengjiyuhong ointment can reduce hypertrophic scars by inhibiting fibroblast proliferation and col agen deposition.
4.Effect and mechnism study of Shengjiyuhong cream on transdifferentiation and collagen production of normal human dermal fibroblasts
Guifang SUN ; Xiaofen ZHANG ; Hongchang LI ; Liyun PAN ; Yafeng CHEN ; Ke XU ; Dianxu FENG
Chinese Journal of Plastic Surgery 2016;32(5):365-372
Objective To explore the effects of Shengjiyuhong cream(SJYHC) on proliferation,transdifferentiation,collagen production and TGF-β1/Smads signaling of normal human dermal fibroblasts (NHDFs).Methods Primary cultured NHDFs between 3-6 passages derived from 6 hypertrophic scar samples were all treated with TGF-β1 (0,2,5,10 ng/ml)stimulation added 5 μg/ml SJYHC or not.After culturing 72 h,CCK-8 solution was added to record absorbance at 450 nm to test proliferation of NHDFs.Fluorescence quantitative PCR was used for testing mRNA expression of α-SMA and type Ⅰ and Ⅲ collagen.Digestion method was to test the hydroxyproline content in the supernatant liquor.Western Blot was used for testing protein expression of α-SMA,type Ⅰ and Ⅲ collagen and Smad2,Smad3,P-Smad2 and P-Smad3.One-way analysis of variance were uesd to analyze differences among more than two groups,while LSD-t test as post hoc test were uesd to make paired-comparisons among the groups.P < 0.05 indicated significant difference.Results With the stimulation of 2,5,10 ng/ml TGF-β1,the absorbance values(A values),mRNA and protein expression of α-SMA and type Ⅰ and Ⅲ collagen,hydroxyproline content,and protein expression of Smad2,Smad3,P-Smad2 and P-Smad3 were all elevated contrasted with the control group (P < 0.05,P < 0.01).Without TGF-β1 stimulation,SJYHC only increased the absorbance values(A values) from 1.645 ±0.052 to 1.796 ±0.060(P <0.05),while mRNA and protein expression of α-SMA and type Ⅰ and Ⅲ collagen,hydroxyproline content,and protein expression of Smad2,Smad3,P-Smad2 and P-Smad3 were infinitely variable(P > 0.05).With stimulation of 2,5 ng/ml TGF-β1,SJYHC elevated the the absorbance values (A values) from 1.814 ± 0.052,1.970 ± 0.045 to 1.981 ± 0.061,2.133 ± 0.059 (P < 0.05).While stimulated with 10 ng/ml TGF-β1,SJYHC declined the absorbance values(A values) from 2.130 ± 0.050 to 1.958 ± 0.045 (P < 0.05).With stimulation of 2,5,10 ng/ml TGF-β1,mRNA expression of α-SMA were declined by SJYHC from 1.04 ±0.06,2.42 ±0.07,7.17±0.11 to 0.28 ±0.06,0.36 ±0.06,1.89 ±0.08 respectively,protein expression from 0.48± 0.05,1.17 ±0.09,2.04 ±0.09 to 0.18 ±0.03,0.21 ±0.08,0.91 ±0.11 respectively (P<0.01),mRNA expression of Col Ⅰ from 0.73 ± 0.08,1.52 ± 0.08,3.05 ± 0.11 to 0.45 ± 0.07 0.46 ± 0.05,1.28±0.09 respectively,protein expression from 0.36 ±0.11,0.94 ±0.10,2.13 ±0.13 to 0.21 ± 0.13,0.24 ±0.08,0.87 ±0.09 respectively (P <0.01),mRNA expression of Col Ⅲ from 1.51 ±0.09,3.28 ±0.09,6.96 ±0.14 to 0.66 ±0.08,0.69 ±0.08,2.23 ±0.10 respectively,protein expression from 0.26 ± 0.08,0.96 ±0.09,1.96 ±0.15 to 0.08 ±0.02,0.12 ±0.02,0.43 ±0.06 respectively (P <0.01),hydroxyproline content from (7.219 ±0.590) μg/ml,(8.745 ±0.514) μg/ml,(10.969 ± 0.489) μg/ml to (6.242 ±0.225) μg/ml,(6.603±0.336) μg/ml,(7.516±0.511) μg/ml (P< 0.05).Under stimulation of 5 ng/ml TGF-β1,SJYHC had no significant effect on protein expression of Smad2 and Smad3 (P > 0.05),while protein expression of P-Smad2,P-Smad3 were all declined from 0.56±0.08,0.87 ±0.13 to 0.31 ±0.07,0.46 ± 0.05 (P <0.01).Conclusions SJYHC may accelerate wound healing and prevent HS by promoting proliferation,inhibiting transdifferation and collagen production and secretion of NHDFs.
5.Effects of Huanglian ointment on wound healing of mice with full-thickness skin defect and the related mechanism
Xiaofen ZHANG ; Guifang SUN ; Yafeng CHEN ; Jiaoyang MA ; Chunfang GAO ; Xia SHENG ; Dianxu FENG
Chinese Journal of Burns 2016;32(12):714-720
Objective To observe the effects of Huanglian ointment on wound healing of mice with full-thickness skin defect,and to explore the related mechanism.Methods Thirty male C57BL/6J mice were divided into Huanglian ointment group and vehicle group according to the random number table after round wounds of full-thickness skin defect with diameter of 7.5 mm were inflicted on the back of each mouse,with 15 mice in each group.Wounds of mice in Huanglian ointment group and vehicle group were treated with Huanglian ointment and vehicle respectively from post injury day (PID) 1 on,2 times each day.Five mice from each group were selected to observe wound changes on PID 0,3,7,10,and 14,and wound healing rates were calculated.Five mice out of the 10 mice that hadn't been used for general observation in each group were sacrificed on PID 3 and 7 respectively,and 5 mice after being used for general observation in each group were sacrificed on PID 14.Wound and skin tissue within 2 mm from the edge of wound was collected.Histologic scoring was conducted based on the histomorphological observation with HE staining.The expression of double positive cells of alpha smooth muscle actin (αt-SMA) and Ki-67 (myofibroblast) in tissue of wounds of mice was observed by immunofluorescence staining.Protein expressions of transforming growth factor beta (TGF-β) and collagen in tissue of wounds of mice were determined by enzyme-linked immunosorbent assay.Data were processed with analysis of variance for repeated measurement,analysis of variance of factorial design,t test of two independent samples,one-way analysis of variance,and Bonferronni test or correction.Results (1) Wounds of mice in two groups were red and swollen on PID 0,while they were neither red nor swollen with scabs on PID 3 and 7.On PID 10,woundsof mice in Huanglian ointment group contracted obviously,while the contracted wounds of mice in vehicle group were smaller than those in Huanglian ointment group.On PID 14,wounds of most mice in Huanglian ointment group were healed,while wounds of some mice in vehicle group failed to heal.Wound healing rates of mice in two groups were close on PID 3 and 7 (with t values respectively 0.64 and 1.90,P values above 0.05).Wound healing rates of mice in Huanglian ointment group on PID 10 and 14 were (76 ±7)% and (93 ±5)% respectively,significantly higher than those of vehicle group [(48 ± 9) % and (68 ± 11) %,with t values respectively 7.44 and 3.89,P values below 0.01].Wound healing rates of mice in two groups on PID 7,10,and 14 were significantly higher than those on the previous time points of the same group (with P values below 0.01).(2) Histologic scores of wounds of mice in two groups were close on PID 3 (t =-0.76,P >0.05).Histologic scores of wounds of mice in Huanglian ointment group on PID 7 and 14 were (7.0 ± 1.6) and (11.6 ± 2.1) points respectively,significantly higher than those of vehicle group [(4.2 ± 1.3) and (7.2 ± 1.3) points,with t values respectively 1.96 and 2.50,P < 0.05 or P < 0.01].Histologic scores of wounds of mice in two groups on PID 7 and 14 were significantly higher than those on the previous time points of the same group (with P values below 0.01).(3) Percentages of double positive cells of α-SMA and Ki-67 in tissue of wounds of mice in Huanglian ointment group on PID 3 and 7 were (35:± 12)% and (62 ± 10) % respectively,significantly higher than those of vehicle group [(17 ± 12) % and (34 ± 6) %,with t values respectively-2.48 and-5.25,P <0.05 or P <0.01].The percentage of double positive cells of α-SMA and Ki-67 in tissue of wounds of mice in Huanglian ointment group on PID 14 was (25 ± 5) %,significantly lower than that of vehicle group [(44 ± 17) %,t =2.50,P < 0.05].The percentage of double positive cells of α-SMA and Ki-67 in tissue of wounds of mice on PID 7 was significantly higher than that on PID 3 or 14 in Huanglian ointment group (with P values below 0.01).Percentages of double positive cells of α-SMA and Ki-67 in tissue of wounds of mice on PID 7 and 14 were significantly higher than those on the previous time points in vehicle group (with P values below 0.05).(4) Protein expressions of TGF-β in tissue of wounds of mice in Huanglian ointment group on PID 3 and 7 were (396 ± 45) and (722 ± 96) pg/mL respectively,significantly higher than those of vehicle group [(290 ± 42) and (382 ± 62) pg/mL,with t values respectively-8.17 and-6.65,P values below 0.01].Protein expressions of TGF-β in tissue of wounds of mice in two groups were close on PID 14 (t =1.60,P > 0.05).The protein expression of TGF-β in tissue of wounds of mice in Huanglian ointment group on PID 7 was significantly higher than that on P1D 3 or 14 (with P values below 0.01).Protein expressions of TGF-β in tissue of wounds of mice in vehicle group on PID 7 and 14 were significantly higher than those on the previous time points (with P values below 0.05).Protein expressions of collagen in tissue of wounds of mice in two groups were close on PID 3 (t =1.99,P > 0.05).Protein expressions of collagen in tissue of wounds of mice in Huanglian ointment on PID 7 and 14 were (47 ± 10) and (70 ± 14) ng/mL respectively,significantly higher than those of vehicle group [(34 ± 10) and (42 ± 12) ng/mL,with t values respectively 3.15 and 3.52,P < 0.05 or P < 0.01].Protein expressions of collagen in tissue of wounds of mice in two groups on PID 7 and 14 were significantly higher than those on the previous time points of the same group (P < 0.05 or P < 0.01).Conclusions Huanglian ointment can promote wound healing of full-thickness skin defect of mice through increasing production of myofibroblasts and protein expressions of TGF-β and collagen.
6.Effect and mechnism study of Shengjiyuhong cream on transdifferentiation and collagen production of normal human dermal fibroblasts
Guifang SUN ; Xiaofen ZHANG ; Hongchang LI ; Liyun PAN ; Yafeng CHEN ; Ke XU ; Dianxu FENG
Chinese Journal of Plastic Surgery 2016;32(5):365-372
Objective To explore the effects of Shengjiyuhong cream(SJYHC) on proliferation,transdifferentiation,collagen production and TGF-β1/Smads signaling of normal human dermal fibroblasts (NHDFs).Methods Primary cultured NHDFs between 3-6 passages derived from 6 hypertrophic scar samples were all treated with TGF-β1 (0,2,5,10 ng/ml)stimulation added 5 μg/ml SJYHC or not.After culturing 72 h,CCK-8 solution was added to record absorbance at 450 nm to test proliferation of NHDFs.Fluorescence quantitative PCR was used for testing mRNA expression of α-SMA and type Ⅰ and Ⅲ collagen.Digestion method was to test the hydroxyproline content in the supernatant liquor.Western Blot was used for testing protein expression of α-SMA,type Ⅰ and Ⅲ collagen and Smad2,Smad3,P-Smad2 and P-Smad3.One-way analysis of variance were uesd to analyze differences among more than two groups,while LSD-t test as post hoc test were uesd to make paired-comparisons among the groups.P < 0.05 indicated significant difference.Results With the stimulation of 2,5,10 ng/ml TGF-β1,the absorbance values(A values),mRNA and protein expression of α-SMA and type Ⅰ and Ⅲ collagen,hydroxyproline content,and protein expression of Smad2,Smad3,P-Smad2 and P-Smad3 were all elevated contrasted with the control group (P < 0.05,P < 0.01).Without TGF-β1 stimulation,SJYHC only increased the absorbance values(A values) from 1.645 ±0.052 to 1.796 ±0.060(P <0.05),while mRNA and protein expression of α-SMA and type Ⅰ and Ⅲ collagen,hydroxyproline content,and protein expression of Smad2,Smad3,P-Smad2 and P-Smad3 were infinitely variable(P > 0.05).With stimulation of 2,5 ng/ml TGF-β1,SJYHC elevated the the absorbance values (A values) from 1.814 ± 0.052,1.970 ± 0.045 to 1.981 ± 0.061,2.133 ± 0.059 (P < 0.05).While stimulated with 10 ng/ml TGF-β1,SJYHC declined the absorbance values(A values) from 2.130 ± 0.050 to 1.958 ± 0.045 (P < 0.05).With stimulation of 2,5,10 ng/ml TGF-β1,mRNA expression of α-SMA were declined by SJYHC from 1.04 ±0.06,2.42 ±0.07,7.17±0.11 to 0.28 ±0.06,0.36 ±0.06,1.89 ±0.08 respectively,protein expression from 0.48± 0.05,1.17 ±0.09,2.04 ±0.09 to 0.18 ±0.03,0.21 ±0.08,0.91 ±0.11 respectively (P<0.01),mRNA expression of Col Ⅰ from 0.73 ± 0.08,1.52 ± 0.08,3.05 ± 0.11 to 0.45 ± 0.07 0.46 ± 0.05,1.28±0.09 respectively,protein expression from 0.36 ±0.11,0.94 ±0.10,2.13 ±0.13 to 0.21 ± 0.13,0.24 ±0.08,0.87 ±0.09 respectively (P <0.01),mRNA expression of Col Ⅲ from 1.51 ±0.09,3.28 ±0.09,6.96 ±0.14 to 0.66 ±0.08,0.69 ±0.08,2.23 ±0.10 respectively,protein expression from 0.26 ± 0.08,0.96 ±0.09,1.96 ±0.15 to 0.08 ±0.02,0.12 ±0.02,0.43 ±0.06 respectively (P <0.01),hydroxyproline content from (7.219 ±0.590) μg/ml,(8.745 ±0.514) μg/ml,(10.969 ± 0.489) μg/ml to (6.242 ±0.225) μg/ml,(6.603±0.336) μg/ml,(7.516±0.511) μg/ml (P< 0.05).Under stimulation of 5 ng/ml TGF-β1,SJYHC had no significant effect on protein expression of Smad2 and Smad3 (P > 0.05),while protein expression of P-Smad2,P-Smad3 were all declined from 0.56±0.08,0.87 ±0.13 to 0.31 ±0.07,0.46 ± 0.05 (P <0.01).Conclusions SJYHC may accelerate wound healing and prevent HS by promoting proliferation,inhibiting transdifferation and collagen production and secretion of NHDFs.
7.Effect of Dachengqi decoction on pancreas aquaporin 1 in rats with acute necrotizing pancreatitis
Yafeng CHEN ; Dianxu FENG ; Teng CHEN ; Jiyun TIAN ; Jinkun XIE ; Haoran SHI ; Jingzhe ZHANG ; Feng HAN
Chinese Journal of Pancreatology 2012;12(1):40-44
Objective To detect the expression of aquaporin 1 in pancreas of rats with acute necrotizing pancreatitis (ANP) and to study the effect of Dachengqi decoction on it.MethodsOne hundred and sixty male SD rats were randomly divided into control group ( C group,n =32 ),ANP group ( n =32),Dexamethasone group (De group,n =32),Acetazolamide group (A group,n =32) and Dachengqi decoction group (DD group,n =32).ANP model was induced by retrograde injection of 5% sodium taurocholate into the biliary and pancreatic duct.Rats in De group received dexamethasone (4 mg/kg) intravenously after ANP induction; while rats in A group received 1 ml acetazolamide via gastric lavage 2 h before ANP induction; rats in DD group received 2 ml Dachengqi decoction via gastric lavage 48,24,2h before ANP induction; rats in C group received laparotomy.Eight rats in each group were sacrificed at 3 h,6 h,12 h and 18h after induction of ANP models.Quantity of ascites and levels of serum amylases were measured.Pathological changes in pancreas tissue were detected by HE and electron microscope.Capillary permeability in pancreas tissue was detected by Evans Blue (EB) extravasations method.AQP1 expression in pancreas tissue was detected by real-time PCR and Western blotting.ResultsLevels of serum amylase in ANP group was significantly higher,and the pancreatic injuries were obvious ; the levels of serum amylase in De group and DD group was lower than that in ANP group,and the pancreatic injuries were attenuated.The levels of serum amylase in A group were higher than that in ANP group,and the pancreatic.injuries were more severe than that in ANP group.Six hours after ANP induction,the levels of EB in pancreas were (13.44 ±2.56),(126.35 ± 14.80),(86.31 ± 14.46),( 108.99 ± 15.07 ),(78.29 ± 16.85 ) mg/L In C group,ANP group,De group,A group and DD group,and the expression of AQP1 mRNA in pancreatic tissue was ( 170.07 ± 22.48 ) %,( 83.93 ± 8.98 ) %,( 117.09 ±10.70 ) %,( 69.00 ± 8.98 ) %,( 112.82 ± 11.79 ) % ; and the expression of AQP1 protein was 0.23 ± 0.06,0.10 ±0.02,0.32 ±0.03,0.13 ±0.02,0.45 ±0.04.The content of EB in ANP group was higher than that in C group,while the expression of AQP1 mRNA and protein in ANP group was significantly lower than that in C group (P < 0.05 ).The content of EB in De group and DD group was significantly lower than that in ANP group,while the expression of AQP1 mRNA and protein was significantly higher than that in ANP group (P < 0.05).ConclusionsAQP1 plays an important role in the pathogenesis of capillary endothelial barrier dysfunction in rats with ANP.Dachengqi Decoction can attenuate pancreatic injuries of rats by regulating the expression of AQP1.
8.Overview in etiology and pathogenesis of hyperlipidemic severe acute pancreatitis
Jinkun XIE ; Yafeng CHEN ; Dianxu FENG
International Journal of Surgery 2012;39(6):416-420
Hyperlipidemia aggravates the pancreatitic injury in the course of severe acute pancreatitis.At present,the research on the pathogenesis has attracted wide attention though it has still not been fully clarified.In many theories of pathogenesis which explained the hyperlipidemic severe acute pancreatitis,the toxic effect of free fatty acids,the theory of accelerated activation of trypsinogen and the theory of pancreatic microcirculation disturbance were the dominant hypothesis.In recent years,concerns have also been raised over the theory of intestinal bacterial.translocation and the theory of calcium overload of pancreatic acinar cells.In this article,the author will review the recent advances in the pathogenesis of hyperlipidemic severe acute pancreatitis.
9.Role of Aquaporin 1 on capillary endothelial barrier dysfunction of intestine in rat with experimental acute necrotizing pancreatitis
Yafeng CHEN ; Dianxu FENG ; Jiyan TIAN ; Teng CHEN ; Peihao YIN ; Jinkan XIE ; Wen FENG ; Junkang GUI
Chinese Journal of General Surgery 2012;27(7):576-581
Objective To study the effect of aquaporin 1 on intestinal capillary endothelial barrier in rats with experimental acute necrotizing pancreatitis (ANP).Methods In this study,160 male Sprague-Dawley rats were randomly divided into five groups:Control group ( n =32),ANP group (n =32),NS group,Dexamethasone group,and Acetazolamide group.Eight rats in each group were sacrificed at 3,6,12 and 18 h after induction of experimental models.Volume of ascites and levels of serum amylases were deternined at each time point.Pathological changes in intestine tissues were observed under electron microscope after HE staining.Capillary permeeabilities in intestine tissues were detected by Evans blue (EB) extravasation experiment.The mRNA and protein expressions of AQP1 in intestine tissue were determined by real-time PCR and Western blotting,respectively.Results Serum amylase level in ANP group was significantly higher than that in control group.Amylase level in dexamethasone group was lower than that in ANP group,and amylase level in acetazolamide group was higher than that in ANP group at 12 h (P <0.05 ) ; The concentration of EB in intestine tissues at each time point in ANP group was significantly higher than those in control group,and EB in dexamethasone group was lower than those in ANP group at 6,12 and 18 h.EB in acetazolamide group was higher than that in ANP group at 3 h ( P < 0.05 ) ; The mRNA expression of AQPI in ANP group was significantly lower than that in control group.The expression of AQP1 in dexamethasone group was higher than those in ANP group at 6,12 and 18 h,and the expression of AQP1 in acetazolamide group was lower than that in ANP group at 3,6,12 h in intestine tissue ( P < 0.05 ).Protein expression of AQPI in tissues in ANP group was significantly lower than that in control group.The expression of AQP1 in dexamethasone group increased more than that in ANP group at 3,6,12 h,and the expression of AQP1 in acetazolamide group was lower than that in ANP group at 3 h,6 h ( P < 0.05 ).Conclusions The expression of AQP1 is down-regulated in intestine tissue in rats with acute necrotizing pancreatitis,and AQP1 could play an important role in the pathogenesis of capillary endothelial barrier dysfunction.
10.The diagnosis and treatment of traumatic spleen rupture: a retrospective analysis of 293 consecutive cases
Dianxu FENG ; Feng HAN ; Shiju ZHU
Chinese Journal of General Surgery 2001;0(08):-
Objective To summarize the experience in the diagnosis and treatment of traumatic spleen rupture.MethodsThe diagnosis and treatment of consecutive 293 patients with traumatic spleen rupture from January 1992 to October 2000 were reviewed.ResultsThe diagnosis was established by the history of injury, clinical presentations, diagnostic peritoneal punctures, abdominal ultrasonography and/or CT. The accuracy rate of diagnosis was 96 3% (282/293). Thirty one patients were treated nonoperatively and cured. Two hundred and fifty of 259 patients treated operatively were cured.Seven had postoperative complications with effusion in the splenic fossa.The total cure rate was 95 9%(281/293).Twelve patients died of uncontrolable hemorrhage or severe multiple injuries.ConclusionsSplenectomy for the treatment of traumatic spleen rupture is satisfactory.The morbidity and mortality of splenectomy are low.

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