1.Effect of Fuling Sini decoction on cardiac function in patients with septic cardiomyopathy
Ping WANG ; Hefei HUANG ; Xiaohong PENG ; Yaxiu HUANG ; Yonglian HUANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2024;31(3):277-282
Objective To explore the effect of Fuling Sini decoction on cardiac function in patients with sepsis-induced cardiomyopathy(SIC).Methods Sixty SIC patients admitted to the department of intensive care unit(ICU)of Shenzhen Hospital(Longgang)of Beijing University of Traditional Chinese Medicine(TCM)from January 2021 to December 2022 were divided into a control group and a treatment group using a random number table method,with 30 patients in each group.Both groups received routine treatment,and the treatment group received Fuling Sini decoction(consisting of Poria cocos 30 g,Dry ginger 12 g,Ginseng 10 g,Prepared aconite 12 g,and Roasted licorice 15 g)based on routine treatment.Each dose was decocted into 200 mL,1 dose per day,divided into 3 times.Both groups of treatments lasted for 8 days.The use time of vasoactive drugs,ICU stay time and total hospital stay time,the 28-day mortality of two groups were recorded.TCM symptom score,sequential organ failure assessment(SOFA),acute physiology and chronic health evaluation Ⅱ(APACHE Ⅱ),procalcitonin(PCT),arterial oxygenation index(PaO2/FiO2),blood lactic acid(Lac),cardiac troponin Ⅰ(cTnⅠ),N-terminal pro-brain natriuretic peptide(NT-proBNP),heart type-fatty acid binding protein(H-FABP),left ventricular ejection fraction(LVEF),left ventricular end-systolic diameter(LVESD),left ventricular end-diastolic diameter(LVEDD),mitral orifice early diastolic blood flow velocity(E),mitral orifice late diastolic blood flow velocity(A),and E/A ratio and tricuspid annular plane systolic excursion(TAPSE)were observed.Results The use time of vasoactive drugs,ICU stay time,and total hospital stay time in the treatment group were significantly shorter than those in the control group[use time of vasoactive drugs(days):4.47±2.16 vs.6.32±3.23,ICU stay time(days):9.18±3.32 vs.12.25±4.39,total hospital stay time(days):13.58±5.14 vs.17.13±6.65,all P<0.05].There was no statistically significant difference in the 28-day mortality between the treatment group and control group[20.00%(6/30)vs.43.33%(13/30),P>0.05].After treatment,the APACHE Ⅱ score and SOFA score in both groups decreased compared to before treatment,with a more significant decrease in the treatment group.The comparison between the two groups was most significant after 8 days of treatment(APACHE Ⅱ score:13.71±3.37 vs.16.21±3.82,SOFA score:3.24±0.85 vs.4.13±1.56,all P<0.05).After treatment,both groups showed a significant decrease in TCM syndrome scores compared to before treatment,with a more significant decrease in the treatment group(26.25±6.44 vs.29.43±6.83 on 3 days of treatment,21.42±4.22 vs.24.81±4.65 on 5 days of treatment,14.43±3.45 vs.17.58±4.56 on 8 days of treatment,all P<0.05).After treatment,PCT,Lac,and H-FABP in both groups decreased compared to before treatment,while PaO2/FiO2 increased,the treatment group showed more significant changes compared to the control group,especially after 8 days of treatment[PCT(μg/L):2.47±1.18 vs.3.54±1.51,Lac(mmol/L):1.86±0.41 vs.2.33±0.64,H-FABP(μg/L):4.67±1.22 vs.6.34±1.55,PaO2/FiO2(mmHg,1 mmHg≈0.133 kPa):297.63±53.92 vs.265.44±48.38,all P<0.05].After treatment,cTnI,NT-proBNP,LVESD,and LVEDD first increased and then decreased in both groups,while LVEF,E/A ratio and TAPSE first decreased and then increased,reaching a valley or peak at 8 days of treatment.Moreover,the above indicators showed statistical significance compared to the control group[cTnI(μg/L):0.15±0.06 vs.0.24±0.13,NT-proBNP(ng/L):825.43±164.73 vs.1234.40±243.37,LVESD(mm):48.36±4.46 vs.52.64±5.15,LVEDD(mm):38.39±3.22 vs.41.87±2.65,LVEF:0.55±0.08 vs.0.50±0.07,E/A ratio:1.23±0.12 vs.1.12±0.08,TAPSE(mm):22.45±2.23 vs.20.55±2.66,all P<0.05].Conclusion Fuling Sini Tang can improve the TCM syndrome of SIC patients,improve heart function,reduce myocardial injury,and shorten hospitalization time,making it a treatment worthy of clinical promotion.
2.Study on the regularity of acupoint selection in the treatment of sepsis with acupuncture and moxibustion based on data mining technology
Yuanyuan ZENG ; Fagen HUANG ; Yaxiu HUANG ; Yonglian HUANG ; Dongjie CHEN ; Boling LI ; Ping WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2024;31(4):407-412
Objective To analyze the acupoint selection rule of acupuncture and moxibustion in the treatment of gastrointestinal dysfunction associated with sepsis,thereby providing a reference for clinical treatment.Methods A systematic search was conducted in several databases,including China national knowledge infrastructure(CNKI),China Science Periodical Database,Chinese Science and Technology Journal Database,Chinese Biomedical literature database(CBM),Wanfang Data,and VIP Database,from the inception of the databases until September 30,2023.The collected literature was organized in an Excel database and subjected to descriptive analysis.Association rule analysis and cluster analysis were performed using SPSS 26.0 and SPSS Modeler 18.0 software.Results Finally,35 acupuncture and moxibustion prescriptions in 35 articles were selected for frequency statistics.The results showed 33 acupoints were used to treat gastrointestinal dysfunction in sepsis,with an overall frequency of use of 176 acupoints.The top 10 acupoints used by acupuncture and moxibustion for the treatment of gastrointestinal dysfunction in sepsis from high to low were Zusanli,Tianshu,Shangjuxu,Zhongwan,Neiguan,Xiajuxu,Guanyuan,Qihai,Shangwan,Xiawan.The top 3 were Zusanli(16.5%),Tianshu(13.6%),Shangjuxu(11.9%).The most commonly used meridians were the Foot Yangming Stomach Meridian(52.8%),Ren Meridian(23.9%),and Foot Taiyin Spleen Meridian(8.0%).The main sites for acupoint selection were the lower limbs(44.9%)and abdomen(41.5%).The most commonly used acupoints were combined acupoints below specific acupoints(35.2%),followed by recruited acupoints(27.8%).The results of high-frequency acupoint correlation analysis showed that the acupoints with the highest comprehensive support were Zusanli,Tianshu,Zhongwan,and Shangjuxu.Cluster analysis identified 5 effective clusters(Quchi acupoint-Sanyinjiao acupoint-Neiting acupoint-Daheng acupoint-Jiaji acupoint-Yinlingquan acupoint-Shangwan acupoint-Xiawan acupoint,Guanyuan acupoint-Qihai acupoint-Neiguan acupoint-Xiajuxu acupoint,Tianshu acupoint-Shangjuxu acupoint,Zhongwan acupoint,Zusanli acupoint)and 4 commonly used acupoint combinations(Guanyuan acupoint-Qihai acupoint-Neiguan acupoint-Xiajuxu acupoint,Tianshu acupoint-Shangjuxu acupoint,Zhongwan acupoint,Zusanli acupoint).Conclusions Acupuncture treatment for gastrointestinal dysfunction in sepsis is mainly based on the treatment of"spleen and stomach",with local acupoint selection and acupoint selection along the meridian as the main approach.It emphasizes the application of specific acupoints,especially the Xiahe and Mu acupoints.
3.Dachaihu decoction protects intestinal mucosal barrier in severe heat stroke rats by inhibiting Toll-like receptor 4/nuclear factor-κB signaling pathway
Dongjie CHEN ; Yonglian HUANG ; Xiaohong PENG ; Yaxiu HUANG ; Fagen HUANG ; Aiting ZHANG ; Ping WANG
Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care 2023;30(5):592-596
Objective To explore the effect of Dachaihu decoction on the Toll-like receptor 4/nuclear factor-κB(TLR4/NF-κB)signaling pathway and gastrointestinal mucosal barrier in rats with severe heat stroke.Methods Sixty SPF grade Sprague-Dawley(SD)male rats were divided into normal control group,model group,Dachaihu decoction standard dose group and Dachaihu decoction high dose group of 15 rats in each group.The heat stroke model was replicated in the rats at temperature(40.5±0.5)℃and humidity(65.0±2.0)%;the normal control group was not treated.From 6 hours after mold making,drug intervention was carried out in the Dachaihu decoction high dose group and the Dachaihu decoction standard dose group of 3.38 g·kg-1·d-1 and 1.69 g·kg-1·d-1,every 8 hours for 2 days.Equal amounts of normal saline were administered to the normal control group and model group.At 6,24 and 48 hours after the molding,5 mL abdominal main artery blood from 5 rats were randomly collected from each group,and the blood was obtained by enzyme-linked immunosorbent assay(ELISA)to determine the levels of tumor necrosis factor-α(TNF-α),interleukin-1(IL-1),D-lactic acid,intestinal fatty acid-binding protein(I-FABP).At the same time,the ileum tissue was retained,and the protein expression of TLR4 and NF-κB in intestinal tissue was determined by Western blotting.Some of the ileal tissue was obtained for hematoxylin-eosin(HE)staining,and the intestinal histopathological changes were observed under light microscopy.Results The normal control group of rats had no significant change,and the other three groups showed heatstroke symptoms after mold making.The overall mortality in drug group were significantly lower than that in the model group[3.3%(1/30)vs.20.0%(3/15),P<0.05].Compared with the normal control group,the serum IL-1,TNF-α,I-FABP,D-lactic acid and the protein expression levels of TLR4 and NF-κB in the model group,Dachaihu decoction standard dose group and Dachaihu decoction high dose group all increased.Compared with the model group,at 24 hours and 48 hours after molding in the Dachaihu decoction standard dose group and Dachaihu decoction high dose group,the serum IL-1,TNF-α,I-FABP,D-lactic acid and the protein expression levels of TLR4 and NF-κB significant decreased[24 hours:TNF-α(ng/L):69.20±4.32,59.37±4.31 vs.76.99±5.02,IL-1(ng/L):132.68±4.93,112.59±9.64 vs.146.75±10.12,I-FABP(mmol/L):504.35±22.23,453.37±32.38 vs.542.58±13.83,D-lactic acid(mmol/L):114.55±8.52,90.57±3.09 vs.127.87±8.37,protein expression of TLR4(A value):1.50±0.08,1.23±0.01 vs.1.86±0.08,protein expression of NF-κB(A value):1.61±0.05,1.21±0.05 vs.1.97±0.08;48 hours:TNF-α(ng/L):58.46±5.13,38.98±5.53 vs.90.21±3.02,IL-1(ng/L):119.12±4.57,84.12±5.08 vs.170.20±6.21,I-FABP(mmol/L):436.04±27.63,321.85±22.03 vs.618.79±12.31,D-lactic acid(mmol/L):87.35±6.84,70.38±4.33 vs.154.14±10.83,protein expression of TLR4(A value):1.19±0.05,1.10±0.13 vs.2.09±0.06,protein expression of NF-κB(A value):1.15±0.09,0.97±0.08 vs.2.20±0.02,all P<0.05].The expression levels of TNF-α,I-FABP,TLR4 and NF-κB protein in Dachaihu decoction high dose group decreased significantly at 24 hours and 48 hours compared with the standard dose group,however,IL-1 and D-lactic acid decreased significantly at 48 hours after molding(all P<0.05).The pathology observation showed that,compared with the model group,the intestinal mucosa villus,the lamina propria drop and haemorrhage was decreased in the Dachaihu decoction standard dose group and Dachaihu decoction high dose group.Telangiectasia was reduced and no ulcer formation was observed.Conclusion Dachaihu decoction can inhibit TLR4/NF-κB signaling pathway,reduce intestinal inflammatory response,thus reduce gastrointestinal damage,and protect the gastrointestinal mucosal barrier in rats with severe heatstroke.
4.Differential expression and significance of Foxp3, TGF-β1 and IL-2 in peripheral blood of patients with coal-burning arsenic poisoning
Xiaolin FANG ; Shiqing XIA ; Kai ZHU ; Ling DONG ; Zhonglan ZOU ; Yonglian LIU ; Dapeng WANG ; Aihua ZHANG
Chinese Journal of Endemiology 2019;38(2):91-95
Objective To investigate the expression of transcription factor forkhead/winged helix transcription factor 3 (Foxp3),immune factor transforming growth factor-beta 1 (TGF-β1),and T-lymphocyte activation related factor interleukin-2 (IL-2) in peripheral blood of patients with coal-burning arsenic poisoning,and to analyze the effects of arsenic exposure on immune function.Methods A case-control study was conducted to investigate 149 cases [94 males and 55 females,(50.69 ± 6.14) years old] of arsenic poisoning in Yuzhang coalburning arsenic poisoning area,southwestern Guizhou Province,and the cases were diagnosed based on the "Diagnosis of Endemic Arsenicosis" (WS/T 211-2015) and confirmed by clinical review.According to skin damage,the patients were divided into mild arsenic poisoning group (39 cases),moderate arsenic poisoning group (54 cases) and severe arsenic poisoning group (56 cases);and 41 cases [12 males and 29 females,(45.76 ± 7.88) years old] of non-arsenic exposed residents from 12 km of Yuzhang coal-burning area were selected as control group.Morning urine and peripheral blood samples were collected with informed consent.Urine arsenic content was measured by inductively coupled plasma mass spectrometry (ICP-MS).Urine arsenic was corrected by creatinine (Cr).Detection of regulatory T cell (Treg)-specific transcription factor Foxp3 gene expression in human peripheral blood was done by real-time fluorescence quantitative PCR,and the levels of Treg-related immune factor TGF-β1 and IL-2 in serum were detected by enzyme linked immunosorbent assay (ELISA).Results The urinary arsenic contents [median (quartile):29.13 (19.75-54.50),31.81 (17.52-53.31),30.51 (18.35-45.76) μg/g Cr] in each arsenic poisoning group were higher than that in the control group [21.62 (17.65-28.44) μg/g Cr,P < 0.05].The expression levels of Foxp3 mRNA in peripheral blood of each arsenic poisoning group [median (quartile):0.58 (0.17-1.27),0.32 (0.17-0.61),0.33 (0.13-0.62)] were significantly lower than that in the control group [0.87 (0.64-1.50),P < 0.05];compared with mild arsenic poisoning group,the expression of Foxp3 mRNA in peripheral blood of moderate and severe arsenic poisoning groups decreased (P < 0.05).The contents of serum TGF-β1 [(13.14 ± 5.19),(12.85 ± 5.51),(12.78 ± 4.95) μg/L] in each arsenic poisoning group were significantly higher than that in the control group [(3.90 ± 1.53) μg/L,P < 0.05].The levels of IL-2 in serum of each arsenic poisoning group [(9.85 ± 5.38),(11.64 ± 6.40),(12.27 ± 6.19) ng/L] were lower than that in the control group [(34.30 ± 4.84) ng/L,P < 0.05];the serum level of IL-2 in severe arsenic poisoning group was higher than that in mild arsenic poisoning group (P < 0.05).Conclusions Arsenic exposure can cause abnormal changes of Treg-specific transcription factor Foxp3 and related immune factors TGF-β1 and IL-2 in peripheral blood of patients.It is suggested that Treg dysfunction may be related to arsenic poisoning.
5.Changes and significance of the ratio of T helper 17 and regulatory T cell in peripheral blood of patients with coal-burning-borne arsenic poisoning
Shiqing XIA ; Xiaolin FANG ; Kai ZHU ; Ling DONG ; Zhonglan ZOU ; Yonglian LIU ; Dapeng WANG ; Aihua ZHANG
Chinese Journal of Endemiology 2019;38(2):101-106
Objective To observe the change of T helper 17 (Th17),regulatory T cell (Treg) as a percentage of lymphocytes and the Th17/Treg ratio in peripheral blood of patients with coal-burning-borne arsenic poisoning,and to explore the role of Th17 cells and Treg cells balance in arsenic-induced immune injury.Methods A case-control study was conducted to investigate 149 cases of arsenic poisoning in Yuzhang arsenic poisoning area in the southwestern Gnizhou Province,and the age was (50.69 ± 6.14) years old,including 94 males and 55 females.The diagnosis was based on the "Diagnosis of Endemic Arsenicosis" (WS/T 211-2015),and the cases were divided into mild arsenic poisoning group (39 cases),moderate arsenic poisoning group (54 cases) and severe arsenic poisoning group (56 cases);forty--one residents of non-arsenic exposed villages about 12 km away from the diseased area were collected as control group,the age was (45.76 ± 7.88) years old,including 12 males and 29 females.Hair samples and peripheral blood were collected from the subjects.The content of hair arsenic was detected by inductively coupled plasma mass spectrometry (ICP-MS).The percentages of Th17 cells and Treg cells in peripheral blood lymphocytes were detected by flow cytometry,and changes in the ratio of Th17/Treg in each group were analyzed.Results The hair arsenic contents in control,mild,moderate,and severe arsenic poisoning groups [median (quartile)] were 0.12 (0.08-0.18),0.20 (0.16-0.33),0.25 (0.18-0.41),0.28 (0.21-0.50) μg/g,and the differences were statistically significant between groups (H =52.22,P < 0.01),and the hair arsenic content in each arsenic poisoning group was higher than that of the control group (P < 0.05).The percentages of Th17 cells in peripheral blood lymphocytes of moderate and severe arsenic poisoning groups [(0.42 ± 0.21)%,(0.41 ± 0.23)%] were higher than that of the control group [(0.29 ± 0.16)%,P < 0.05].The percentages of Treg cells in peripheral blood lymphocytes of mild,moderate and severe arsenic poisoning groups [(0.37 ± 0.18)%,(0.31 ± 0.19)%,(0.27 ± 0.18)%] were lower than that of the control group [(0.71 ± 0.20)%,P < 0.05];with respect to the mild arsenic poisoning group,the percentage of Treg cells in severe arsenic poisoning group was reduced (P < 0.05).The ratios of Th17/Treg in mild,moderate and severe arsenic poisoning groups (1.17 ± 0.63,1.56 ± 0.69,1.83 ± 0.85) were higher than that of the control group (0.43 ± 0.22,P < 0.05);compared with mild arsenic poisoning group,the ratio of Th17/Treg in severe arsenic poisoning group was elevated (P < 0.05).Correlation analysis showed that the hair arsenic content was positively correlated with the percentage of Th17 cells in peripheral blood lymphocytes and the ratio of Th17/Treg (r =0.323,0.608,P < 0.05),and negatively correlated with the percentage of Treg cells in peripheral blood lymphocytes (r =-0.486,P < 0.05).Conclusion Coal-burning-borne arsenic poisoning can cause the proportion of Th17 cells in the peripheral blood of patients to increase in lymphocytes,and the proportion of Treg cells in lymphocytes to decrease,which in turn changes the balance of Th17/Treg,resulting in weakened immune tolerance and disorder the regulation of inflammatory response,thus participates in the occurrence and development of arsenic-induced immune damage.
6.miR-1180-5p inhibits proliferation, migration and invasion of prostate cancer cells by activating CDKN1Agene expression
WANG Yong ; GUO Yonglian ; CHEN Lin ; LI Guohao ; Ying Chengcheng ; CHENG Wei
Chinese Journal of Cancer Biotherapy 2018;25(7):698-703
Objective: To study the effects of microRNA-1180-5p (miR-1180-5p) on malignant biological behaviors of prostate cancer VCAP and LNCaP cells and the possible mechanisms. Methods: dsControl (dsControl group) and miR-1180-5p (miR-1180-5p group) were constructed and then transfected into two prostate cancer cell lines VCAPand LNCaP. qPCR and Western blotting were used to analyze the changes in mRNA and protein expressions of CDKN1A, Cyclin D1 and CDK6 after transfection. Cell cycle distribution, proliferation activity, clone formation capacity, cell migration and invasion ability were detected by flow cytometry, MTT assay, colony culture assay and Transwell assay, respectively. Results: qPCR results showed that compared with dsControl, CDKN1A mRNA levels in VCAP and LNCaP cells transfected with miR-1180-5p were up-regulated significantly, while the mRNA expressions of Cyclin D1 and CDK6 were significantly down-regulated (all P<0.01). Western blotting result was consistent with that of qPCR. The percentage of cells in G0/G1 phase was increased after transfection of miR-1180-5p (P<0.05), but the proportion of cells in S phase and G2/M phase was decreased and the cell cycle was arrested at G0/G1 phase (P<0.05). The proliferation activity of the two prostate cancer cells was significantly lower than that of the dsControl group after miR-1180-5p transfection (P<0.05), and the number of colonies in the miR-1180-5p group was significantly lower than that in the dsControl group (P<0.01). In the meanwhile, the cell migration and invasion ability in miR-1180-5p group was decreased (P<0.01). Conclusion: miR-1180-5p can significantly activate CDKN1A gene expression in prostate cancer cells and further inhibit the proliferation, migration and invasion of prostate cancer cells.
7.Differential expression and significance of CD4+CD25+Foxp3+regulatory T cell in liver of arsenic-exposed rats
Ling DONG ; Aihua ZHANG ; Kai ZHU ; Dapeng WANG ; Yonglian LIU ; Zhonglan ZOU ; Qingling WANG
Chinese Journal of Endemiology 2018;37(2):96-101
Objective To observe the differential expression level of CD4+CD25+Foxp3+regulatory T cells (Treg) in liver of arsenic-exposed rats, explore the regulatory mechanisms on immunological of hepatic injury induced by arsenic, and provide a basis for prevention and treatment of the disease. Methods Thirty-two healthy Wistar rats were selected and randomly divided into control,low,medium and high arsenic dose groups by weight,8 rats per group. Rats in control group were given oral gavage of deionized water, while the other groups were given oral gavage doses of 2.00 g/L sodium arsenite(NaAsO2) according to their body weight for 6 days every week, the concentrations were 1.25, 2.50 and 5.00 ml/kg. After 4 months, liver tissue samples of rats were collected, the content of arsenic in liver was detected by inductively coupled plasma mass spectrometry (ICP-MS);the expression of Treg cells in liver was detected by immunohistochemistry; enzyme-linked immunosorbent assay (ELISA) was applied to detect the levels of interloukin-10 (IL-10),transforming growth factor beta 1 (TGF-β1), IL-6, IL-17 and IL-2. Results Compared with the control group [28.57 (17.64 - 35.64)μg/g], the content of arsenic in liver in low,medium and high arsenic exposed groups[M(P25-P75):638.30(527.91-802.58),591.64(513.82-723.16),792.55 (695.93 - 1 074.41) μg/g] increased, the differences were statistically significant(P < 0.05). Compared with low arsenic group, the content of arsenic in liver in high arsenic group increased, the difference was statistically significant (P < 0.05). Numerical density on area (NA) of positive Treg cells in medium,high arsenic exposed groups [(2.25 ± 0.50),(4.00 ± 2.16)A/cm2]was higher than that of the control group[(0.60 ± 0.54)A/cm2,P<0.05];NA of positive Treg cells in high arsenic exposed group was higher than that of the low arsenic exposed group[(1.50 ± 0.58) A/cm2, P < 0.05]. The expressions of the IL-10 in low, medium and high arsenic exposed groups [(5.58 ± 1.70), (6.78 ± 1.09),(7.18 ± 0.53)μg/L]were higher than that of the control group[(2.32 ± 0.83) μg/L,P<0.05];compared with low arsenic group, the expression of IL-10 in high arsenic group increased (P < 0.05); compared with control group [(1.46 ± 0.65) μg/L], the expression of TGF-β1 in high arsenic exposed group increased[(9.06 ± 3.60)μg/L, P<0.05];compared with control group [(2.33 ± 0.66)μg/L], the expression of IL-6 in high arsenic exposed group increased [(5.03 ± 1.39) μg/L, P < 0.05], compared with low arsenic exposed group [(2.46 ± 1.71) μg/L], the expressions of IL-6 in high arsenic exposed group increased, the difference was statistically significant (P < 0.05);the expression of IL-17 among control, low, medium and high arsenic exposed groups[(4.87 ± 1.64),(7.50 ± 2.74), (6.21 ± 1.47),(7.23 ± 2.68)μg/L]were not statistically significant (F = 1.429, P > 0.05); compared with control group [(16.30 ± 3.98) μg/L], the expression of IL-2 in high arsenic exposed group decreased[(9.93 ± 2.65) μg/L, P <0.05]. The content of arsenic in liver was positively correlated with the expression of IL-10, TGF-β1, IL-17, IL-6 (rs=0.696,0.463,0.632,0.502,P<0.05),and negatively correlated with the expression of IL-2(rs=-0.522,P<0.05). Conclusion With increasing of arsenic exposure level, the content of arsenic in liver and the expression of CD4+CD25+Foxp3+Treg have increased,the cytokines are secreted abnormally,liver immunological micro environment is disordered,immune tolerance is formed,and immune clearance is inhibited,which may play an important role in the occur and development of immunological liver damage induced by arsenic in rat.
8.Roles of T helper 17 and regulatory T cell infiltration in hepatic injury induced by arsenic in rats
Kai ZHU ; Aihua ZHANG ; Ling DONG ; Dapeng WANG ; Yonglian LIU ; Zhonglan ZOU ; Qingling WANG
Chinese Journal of Endemiology 2018;37(7):536-540
Objective To investigate the infiltration of T helper 17 (Th17) and regulatory T cells (Treg) in the liver of rats exposed to arsenic,and to investigate the roles of Th17 and Treg in infiltration of liver injury induced by arsenic.Methods Thirty-two Wistar rats,half male and half female,were randomly divided into control,low,medium and high arsenic dose groups by body weight via the random number table method,8 rats per group.Rats in control group were given oral garage of deionized water,while other groups were given oral gavage doses of 2.00 g/L sodium arsenite (NaAsO2) according to their body weight for 6 days every week,the concentrations of NaAsO2 were 1.25,2.50 and 5.00 ml/kg,respectively.After 4 months,liver tissue samples of rats were collected,the content of arsenic in liver was determined by inductively coupled plasma mass spectrometry (ICP-MS);Hematoxylin-eosin staining (HE) method was used to observe the morphological changes of liver tissue in rats;the protein expressions of interleukin-17A (IL-17A,the imflammatory factor secreted by Th17 cells) and Forkhead Box P3 (Foxp3,the lineage-specific transcription factor of Treg cells) were measured with immunohistochemistry.Results ① Arsenic content in liver of low,medium,and high arsenic exposed groups [63.83 (52.79-80.26),59.16 (51.38-76.58),79.26 (69.59-107.44) μg/g] were higher than those of the control group [2.86 (1.76-3.56)μg/g,P < 0.05],and the high arsenic dose group was higher than the medium arsenic dose group (P < 0.05).② With increasing doses of arsenic,the numbers of inflammatory cells in the liver tissue of rats were increased,and the liver tissue of the high arsenic dose group showed vacuolar degeneration and pathological changes in some areas.③ Compared with the control group,low and medium arsenic dose groups (0.001 + 0.001,0.010 ± 0.020,0.030 ± 0.080),the expression of IL-17A protein in the liver in high arsenic dose group were significantly increased (0.220 ± 0.130,P < 0.05),the differences were statistically significant between groups (F =14.776,P <0.05).The expressions of Foxp3 protein in the liver in low,medium,and high arsenic dose groups were significantly higher (0.270 ± 0.050,0.330 ± 0.040,0.320 ± 0.070) than that in the control group (0.070 ± 0.020),the differences were statistically significant between groups (F =56.990,P < 0.05).④ There was a positive correlation between hepatic arsenic levels and protein levels of IL-17A and Foxp3 in liver (r =0.48,0.81,P < 0.05).Conclusion Arsenic exposure can increase the content of arsenic in liver tissue of rats,which causes the changes of infiltration of Th17 and Treg cells,leading to the change of immune status,suggesting that Thl7 and Treg cells play an important role in the development of arsenic-induced immune injury.
9.Effect of microRNA-1180 transfection on growth of renal cell carcinoma
Yong WANG ; Yonglian GUO ; Lin CHEN ; Guohao LI ; Jiajun YU ; Wei CHENG
Chongqing Medicine 2018;47(6):736-739
Objective To investigate the effect of microRNA-1180 transfection on the growth of renal cell carcinoma lines 786-O and ACHN.Methods The renal carcinoma cells were divided into the two groups:control group (transfecting dsControl) and experimental group (transfecting miR-1180).The expression change of p21 mRNA was detected by qRT-PCR.Western blot was conducted to analyze the expression changes of p21,CDK4,CDK6 and CyclinD1 proteins.Flow cytometry (FCM) was used to detect the cell cycle change.The MTS assay was conducted to detect the cell viability and the colony forming assay was performed to examine the cell proliferation ability.Results The qRT-PCR results showed that compared with the negative control dsControl group,after miR-1180 transfection,the expression level of·p21 mRNA in 786-O and ACHN cells was up-regulated to 2.54-fold and 2.49-fold respectively(P<0.01).The expression trend of p21 protein was consistent with qRT-PCR results.The expression of CDK4,CDK6 and CyclinD1 proteins were significantly down-regulated.The FCM results showed that the proportion of cells in G0/ G1 phase was significantly increased after transfection of miR-1180,but the proportion of cells in S phase and G2/M phase was decreased significantly,indicating that the cell cycle was arrested in G0/G1 phase.The MTS assay results showed that compared with the dsControl group,the viability of the two kinds of renal carcinoma cells was significantly decreased.The colony formation assay showed that the number of colonies formed in the miR-1180 group was smaller,indicating the proliferation ability of miR-1180 transfected cells was decreased.Conclusion miR-1180 can significantly activate the p21 protein expression and inhibit the growth of renal carcinoma cell lines 786-O and ACHN.
10.Effect of exogenous small RNA transfection on the growth of renal clear cell carcinoma cells
Yong WANG ; Yonglian GUO ; Lin CHEN ; Guohao LI ; Jiajun YU ; Wei CHENG
The Journal of Practical Medicine 2017;33(10):1572-1575
Objective To investigate the effect of exogenous small RNA (dsP21-397) transfection on growth of human renal clear cell carcinoma cell lines A-498 and Caki-1. Methods The dsControl(control group) and dsP21-397(experimental group)were transfected into A-498 and Caki-1,respectively. The expression of p21 mRNA was analyzed by qRT-PCR. The expression of p21 protein,CDK4 and Cyclin D1 proteins were detected by Western blot. The cell cycle distribution was examined by flow cytometry(FCM). MTS assay and colony formation assay were used to analyze cell viability and proliferation ability. Results Compared with dsControl,p21 mRNA levels in A-498 and Caki-1 cells increased to 2.55-fold(P<0.01)and 2.18-fold(P<0.01),respectively,after transfection with dsP21-397. The expression of p21 protein was up-regulated while the expression of CDK4 and CyclinD1 were down-regulated. The percentage of cells in G0/G1 phase significantly increased after transfection of dsP21-397,and the proportion of cells in S phase and G2/M phase significantly decreased. The activity of A-498 and Caki-1 cells significantly decreased and the number of colonies in the dsP21-397 group was significantly lower. Conclusions dsP21-397 can significantly activate p21 protein expression,down-regulate the cell cycle-associated proteins,and inhibit the growth of renal clear cell carcinoma cells.

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