1.Mechanism of Qingre Huayu Jianpi Prescription Inhibiting Development of Colitis-associated Colorectal Cancer in Mice
Yanqiu ZHENG ; Yihui YOU ; Junyu KE ; Jinbin SONG ; Yongqiang WU ; Changhui LIU ; Yanwu LI
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(8):83-90
ObjectiveTo explore the effect of Qingre Huayu Jianpi prescription (QHJ) on colitis-associated colorectal cancer (CAC) in mice, and its related mechanism. MethodC57BL/6 mice were randomly divided into four groups including the normal, model, QHJ low-dose (QHJ-L, 10 g·kg-1), and QHJ high-dose (QHJ-H, 40 g·kg-1) groups. Azoxymethane (AOM) and dextran sodium sulfate (DSS) were combined to chemically build a CAC mouse model for 14 weeks. Each drug group was given intragastrically from the 5th week to the 14th week, once per day. An equal volume of water was fed to the normal and model groups. The mouse survival rate, colon length, weight, and pathological alterations were assessed. The protein expressions of Wnt-3a protein signaling (Wnt3a), β-catenin, Non-phosphor-β-catenin (Non-p-β-catenin), and cholesterol-binding glycoproteins 133 (CD133) were detected by Western blot. The localization and expression of the cluster of differentiation (CD) 80 and CD11 antigen-like family member B (CD11b) were detected by immunohistochemistry (IHC). The colon organoids derived from CAC mice were isolated and cultured to detect the expression of Wnt signaling pathway-related proteins. ResultThe survival rate of the CAC mice was improved by QHJ treatment and the number of colon tumors was inhibited significantly. Compared with those in the normal group, the expression levels of Wnt3a, β-catenin, Non-p-β-catenin, and CD133 in colon tissues in the model group were significantly increased (P<0.05, P<0.01). Compared with those in the model group, the levels of Wnt3a and β-catenin in the QHJ-L group were significantly decreased (P<0.01), and the protein levels of Wnt3a, β-catenin, Non-p-β-catenin, and CD133 in the QHJ-H group were significantly decreased (P<0.05, P<0.01). Meanwhile, the expression level of CD11b in the model group was significantly increased compared with that in the normal group while the CD80 level was significantly decreased (P<0.05, P<0.01). Compared with those in the model group, CD11b in QHJ-L and QHJ-H groups was significantly decreased, and CD80 was significantly increased(P<0.05, P<0.01). The expressions of Non-p-β-catenin and CD133 in colonic organoids of CAC model mice were significantly increased, while QHJ treatment could inhibit the expressions of Non-p-β-catenin and CD133 in colonic organoids (P<0.01). ConclusionQHJ could inhibit the inflammation-cancer development in CAC mice, the mechanism of which might be related to regulating the microenvironment and inhibiting the over-activation of Wnt signaling.
2.Effect of Buzhong Yiqiwan on NLRP3 Inflammasome Pathway of DSS-induced Colitis Model Mice at Different Pathological Stages
Chunhui SONG ; Yihui YOU ; Junyu KE ; Geng LI ; Haishan LONG ; Yanli WU ; Qun DU ; Yanwu LI ; Wenfeng GUO
Chinese Journal of Experimental Traditional Medical Formulae 2022;28(14):20-28
ObjectiveTo explore the intervention effect and mechanism of Buzhong Yiqiwan (BZYQ) on colitis mice. MethodSixty-four C57BL/6 mice were randomly divided into 2 weeks blank group, 2 weeks model group, 2 weeks high-dose BZYQ (12 g·kg-1) group, 2 weeks low-dose BZYQ (6 g·kg-1) group, 4 weeks blank group, 4 weeks model group, 4 weeks high-dose BZYQ (12 g·kg-1) group, and 4 weeks low-dose BZYQ (6 g·kg-1) group. The colitis model was induced in mice by feeding 3% dextran sodium sulfate (DSS) for 7 days. The mice received BZYQ (12 and 6 g·kg-1) by gavage on the 8th day after modeling, once per day, and sacrificed on the 2nd and 4th weeks, correspondingly. The colon length and weight of mice in each group were measured. Hematoxylin-eosin (HE) staining was used for pathological observation and colonic mucosal inflammation was scored. The mRNA and protein expression of NOD-like receptor thermoprotein domain 3 (NLRP3), apoptosis-associated speck-like protein containing a CARD (ASC), and cysteinyl aspartate-specific protease-1 (Caspase-1) was detected by real-time quantitative polymerase chain reaction (Real-time PCR) and Western blot, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of inflammatory cytokines, such as interleukin (IL)-1β, IL-18, and IL-33 in colonic tissues. ResultCompared with the 2 weeks blank group, the 2 weeks model group showed shortened colon length, increased colon weight (P<0.05), loss of epithelial cells, destruction of gland structure, infiltration of a large number of inflammatory cells in mucosa and submucosa, local crypt abscess, and increase in mucosal inflammation score (P<0.01) as revealed by light microscopy, elevated levels of IL-1β, IL-18, and IL-33 in colonic tissues (P<0.05), and increased mRNA and protein expression of NLRP3, ASC, and Caspase-1 (P<0.05). The intervention of BZYQ (12 g·kg-1) restored colon length, alleviated mucosal injury (P<0.05), down-regulated the content of IL-18 (P<0.05), reduced the mRNA expression of NLRP3 and ASC as well as the protein expression of ASC and Caspase-1 compared with the conditions in the 2 weeks model group. Compared with the 4 weeks blank group, the 4 weeks model group showed decreased colon length, increased colon weight (P<0.05), decreased glands in the mucosal layer, expansion of glandular cavity, atrophy of crypt, local connective tissue hyperplasia and lymphocyte infiltration, increased inflammation score (P<0.01) as revealed by the light microscopy, increased expression of IL-1β, IL-18, and IL-33 (P<0.05), and elevated mRNA and expression of NLRP3 inflammasome complex (P<0.05). Compared with the conditions in the 4 weeks model group, the intervention of BZYQ (12 and 6 g·kg-1) could improve colon length and weight (P<0.05), and the intervention of BZYQ (12 g·kg-1) could also improve the inflammation score of the colon (P<0.05). Different from the acute stage, the intervention of BZYQ (12 and 6 g·kg-1) increased the content of IL-33 in the intestinal mucosa and up-regulated the mRNA and protein expression of NLRP3 inflammasome complexes ASC and Caspase-1 (P<0.05). ConclusionBZYQ can relieve the injury of colitis induced by DSS in mice. The mechanism is related to the regulation of intestinal immune response mediated by NLRP3 inflammasome, and it has different regulatory effects in acute and chronic inflammation stages.
3.Effects of astragaloside on TRPC6 expression on mouse podocyte induced by TGF-β1
Haiting HUANG ; Haohao WU ; Youling QIN ; Xu LIN ; Yanwu YOU ; Pengwei GUO ; Chunrong TANG
Chinese Journal of Immunology 2017;33(3):370-373
Objective:To explore the possible mechanism of astragaloside involved in the mouse podocytes injury induced by TGF-β1 in vitro.Methods:Mouse podocytes were cultured in vitro and then all cell were divided into 5 groups:normal control group , TGF-β1 treatment group ,TGF-β1 treatment +astragaloside low dose group ,TGF-β1 treatment +astragaloside middle dose group and TGF-β1 treatment +astragaloside high dose group.The proliferation rate of each group was investigated by MTT assay ,the expression of TRPC6 protein and mRNA were measured by Western blot and RT-PCR respectively after 48 hours.Results:TGF-β1 can significantly inhibit the proliferation of podocytes ( P<0.05) ,fusions of foot processes or even effaced of podocytes were observed .TGF-β1 could also increase the expression of TRPC6.Astragaloside could reduce the inhibition of TGF-β1 to the proliferain of podocytes significantly ,make the cell shape tend to be normal,and reduce the expression of TRPC6 mRNA and protein with dose-effect relation.Conclusion:TRPC6 play an impor-tant role in the TGF-β1 induecd podocytes injury .Astragaloside can alleviate podocytes injury by reduce the expression of TRPC 6.
4.Observation on effect of fentanyl combined with midazolam in painless bronchoscopy
Chinese Journal of Biochemical Pharmaceutics 2017;37(5):321-322,325
Objective To evaluate the effect of fentanyl combined with midazolam in painless bronchoscopy.Methods76 patients underwent painless bronchoscopy in our hospital from October 2015 to October 2016 were randomly devided into the observation group and the control group, 38 cases in each group.The control group received conventional lidocaine local anesthesia, based on the control group, the observation group were given intravenous anesthesia using fentanyl and midazolam.Hemodynamic parameters (blood oxygen saturation (SpO2), heart rate (HR), diastolic blood pressure (DBP), systolic blood pressure (SBP)) were compared in the two groups before anesthesia (t1), 3 minutes after anesthesia (t2), 5 min after anesthesia (t3), wake up (t4), besides, incidence of adverse reactions were compared as well.ResultsAt t2, t3 time point, SpO2 was higher in the observation group than that in the control group, HR, DBP and SBP were lower than the control group, the difference was statistically significant (P<0.05);the incidence of side effects in the observation group was 5.26% (2/38), which was lower than that in the control group (26.31%, 10/38), the difference was statistically significant (P<0.05).ConclusionFentanyl combined with midazolam in painless bronchoscopy can effectively maintain the patient's hemodynamic stability and reduce the incidence of adverse reactions.
5.Establishment of a rhesus monkey model of chronic temporal lobe epilepsy
Yajie CHI ; Shan XUE ; Haitao SUN ; Yuting XU ; Bolin WU ; Jianwei GUAN ; Jin'e YUAN ; Zhiming LIANG ; Zhaohui WU ; Qiang XU ; Zhaoming LIU ; Liangxue LAI ; Yanwu GUO
Chinese Journal of Neuromedicine 2017;16(6):564-569
Objective To describe a rhesus monkey model of temporal lobe epilepsy (TLE) established via repetitive unilateral intra-amygdala kainic acid (KA) injection and provide experimental basis for epileptogenic network and related research. Methods Eight male adult rhesus monkeys were randomly divided into KA injection group (n=6) and saline injection group (n=2). Brain stereotaxic technique, micro catheter implantation into the right amygdaloid nucleus, subcutaneous bladder connection, and continuous video-EEG monitoring were performed, and KA or saline injection into their right amygdala was achieved. Interictal epileptic discharges (IEDs), ictal discharges and behavioural performance between the two groups were compared right after injection and within 6 months of first discharge. Results Typical IEDs were recorded in the 6 monkeys from KA injection group after 2-4 times of KA injection, with focal spike waves discharges at the right temple area as manifestation; ictal discharges were recorded in 4 monkeys, with discharge patterns of discharges from the right temple area to the whole brain as manifestation, and during epileptic attack, these 4 monkeys suddenly stopped and dumbfounded without obvious limb seizures. Monkeys from the saline injection group showed no obvious abnormal behaviors. Conclusion Through a modified protocol of unilateral repetitive intra-amygdala KA injection, a rhesus monkey model with high similarity of behavioral and brain electrical features to TLE is developed.
6.Effect of human amniotic mesenchymal stem cell transplantation on motor behaviors in hSOD1-G93A mouse models of amyotrophic lateral sclerosis
Kuntai XIAO ; Xiaomei LIANG ; Ya GAO ; Mubin CHEN ; Duobin WU ; Xiaodan JIANG ; Yanwu GUO ; Haitao SUN
Chinese Journal of Neuromedicine 2017;16(7):649-656
Objective To explore the therapeutic effect of intravenous transplantation of human amniotic mesenchymal stem cells (hAMSCs) on protection of motor behaviors in hSOD1-G93A mouse models of amyotrophic lateral sclerosis (ALS).Methods Amnion membranes were obtained from placentas delivered by healthy mother donors.The hAMSCs were gradually isolated and purified from amnion membranes using tissue culture method.Immunophenotypes of the isolated hAMSCs were analyzed using fluorescence activated cell sorter (FACS).Transgenic mice harboring a high copy number of hSOD1-G93A (B6SJL-TgN [SOD1-G93A] 1Gur) transgene were used in this study.Hemizygous transgenic progenies were maintained by mating the transgenic males with F1 hybrid wild-type (WT)females.The progenies were genotyped by polymerase chain reaction (PCR) using genomic DNA isolated from mouse tail after birth.The study included hSOD1-G93A mice transplanted with hAMSCs,PBS-injected transgenic mice,and normal WT mice (n=12).The hAMSCs were administered intravenously in jugular vein of the mice under anesthesia.The cells (1 ×106) in 200 μL PBS were delivered over 10 min.Animals received cells or PBS at 12,14,and 16 weeks old,respectively.The disease onset and progression of ALS mice models were monitored using rotarod performance test,PaGE test,and CatWalk gait analysis since 8 weeks old every week.Results (1) The immunophenotype of the isolated hAMSCs was conformed using FACS.These cells were positive for CD29,CD44,CD73,CD90,and CD166,but negative for CD14,CD34,CD45,CD123,and human leukocyte (site) DR antigen.Interestingly,stage specific embryo surface antigen 4 and octuber binding transcription factor 4 were detected in hAMSCs.(2) ALS mice in the hAMSCs transplantation group had significantly improved motor functions than those in the PBS treatment group:motor performance on the rotarod test (from 14 to 18 weeks old),PaGE test (from 15 to 18 weeks old) and CatWalk gait analysis (from 15 to 19weeks old) in hAMSCs-injected ALS mice was significantly improved as compared with that in the PBS treatment group (P<0.05).Conclusions The multiple transplantation of hAMSCs by intravenous delivery can bring amelioration of the disease phenotype,as evidenced by improved motor function in hSOD1-G93A mouse models.The hAMSCs transplantation can be considered as a promising cellular treatment for ALS.
7.Inhibitory Effects of Pharbitidis Semen on Rat Hapatoma Induced by N-nitrosodiethylamine
Rongmin WU ; Xiaoyan FANG ; Yanwu LING ; Biao LI ; Binchen HUANG
Herald of Medicine 2015;(4):463-466
Objective To study the inhibitory effect of Pharbitidis Semen on rat hepatoma induced by N-nitrosodiethylamine ( NDEA) . Methods SD rats were divided into normal control group, model control group and Pharbitidis Semen group. In model control group and Pharbitidis Semen group, 0. 01% NDEA was applied for 90 days to induce hepatoma, and rats in Pharbitidis Semen group concomitantly received feed containing 6% Pharbitidis Semen at the dosage of 40 g·kg-1 ·d-1 . Thirty days after the hepatoma inducement and Pharbitidis Semen administration, the rats were sacrificed to observe the pathological changes in liver, number of hepatoma nodules and liver weight. The changes of liver/body weight, serum alanine aminotransferase (ALT), γ-glutamyl transferase (γ-GT), and alkaline phosphatase (ALP) were compared. One-way ANOVA (LSD Test) was employed for statistical analysis. Results In the normal control group, the number of hepatoma nodules was 0. 0±0. 0, the liver weight was (9. 87±1. 30) g, the ratio of liver/body weight was (2. 62±0. 24)% and the level of serum ALT was (64. 10±12. 71) U·L-1,γ-GT was (0. 80± 0. 42) U·L-1, and ALP was (121. 20±37. 57) U·L-1. In the model control group, the number of hepatoma nodules was (27. 4±9. 5), the liver weight was (21. 38±7. 29) g, the ratio of liver/body weight was (5. 82±2. 31)%, the level of serum ALT was (175. 70±48. 75) U·L-1, γ-GT was (41. 80±15. 38) U·L-1, and ALP was (200. 50±35. 78) U·L-1. In the Pharbitidis Semen group, the number of hepatoma nodules was (8. 6± 5. 3), the liver weight was (13. 91±3. 55) g, the ratio of liver/body weight was (3. 86±0. 76)% and the level of serum ALT was (113.10±45.35) U·L-1, γ-GT was (13. 40± 6. 15) U·L-1, and ALP was (155. 80±30. 26) U·L-1. The results showed that all indices of Pharbitidis Semen group were higher than those of the normal control group, and lower than those of the model control group (P<0. 01 or P<0. 05). Conclusion Pharbitidis Semen can reduce NDEA-induced injury to the liver cells, and inhibit the overgrowth of the hepatoma.
8.Antidiarrheal and analgesic effects of Dingguier paste on the mouse models
Yuhong YU ; Guowei ZHANG ; Xiaoning YANG ; Yanwu LI ; Yanli WU ; Rujun WANG
International Journal of Traditional Chinese Medicine 2014;(6):535-538
Objective To observe the antidiarrheal and analgesic effects of Dingguier paste on the mouse. Methods Tho diarrhea mouse models were made by senna leaf or castor oil to detect the antidiarrheal effect of Dingguier paste combination with other drugs. The hot plate test and acetic acid test were used to observe the analgesic effect of Dingguier oral medicine and Dingguier paste in combined use with other drugs. Results Compared with senna leaf model group, Dingguier paste low-dose, middle-dose, high-dose combined with diphenoxylate, Bupi-Yichang pill and Dingguier oral medicine group could decrease the number of feces in 2 h [Number of each group feces was(0.50±1.27), (2.27±2.90), (1.18±1.83);(2.00±2.26), (3.40±2.17), (3.82±1.72);(0.73±1.56), (1.91±2.95), (2.55±2.11)]. Dingguier paste low-dose and high-dose combined with diphenoxylate could decrease the number of feces in 4h [Number of each group feces was (6.70±2.11), (6.27±3.20)]. Dingguier oral group decrease the number of feces in 6h [Number of feces was (6.91±2.77)]. Compared with castor oil model group, Dingguier paste low-dose combined with diphenoxylate could obviously decrease the number of feces in 2 h, 4 h and 6 h [Number of each group feces was(0.45±0.82), (0.45±0.82), (4.27±2.15)]. Dingguier paste middle-dose combined with diphenoxylate could decrease the number of feces in 6 h [Number of feces was (4.64±2.45)]. Dingguier oral group decrease the number of feces in 2 h, 4 h, 6 h [Number of each group feces was (0.45±0.82), (2.91±2.07), (4.27±2.15)].Compared with normal group, the threshold of pain stimulated by hot plate of the Dingguier paste each dose combined with rotundin could increased remarkable after 30 min, 60 min, 90 min and 120 min[The time of Dingguier paste low-dose was(52.32 ± 17.29)s, (56.24 ± 12.48)s, (57.28 ± 6.61)s, (58.58 ± 4.70)s; the time of Dingguier paste middle-dose was (48.27±19.35)s, (54.92±12.59)s, (55.91±11.48)s, (55.15±12.49)s;the time of Dingguier paste high-dose was (49.15±15.69)s, (56.24±11.89)s, 60 s is(59.08±2.93)s]. Compared with normal group, the eclipse time of pain irritated by acetic acid of the Dingguier paste middle-dose combined with indomethacin could increased remarkable(14.72±5.99)min, the writhing frequency could decrease of the Dingguier paste each dose combined with indomethacin[Number of each dose writhing frequency was (7.62±8.31), (3.62± 6.14), (11.25±9.46)]. Conclusion Dingguier paste in the combination use with other drugs and Dingguier oral medicine may have some antidiarrheal and analgesia effects.
9.Measurement of the angular velocity and perpendicular ground reaction force of the ankle joint in parachute landing simulation
Chao ZHENG ; Ji WU ; Rongrong HUANG ; Songchao CUI ; Yanwu WEN ; Yi LI ; Di WU
Chinese Journal of Orthopaedics 2014;34(6):688-693
Objective To measure the angular velocity and perpendicular ground reaction force of the ankle joint under different heights with half-squat jumping in parachute training simulation,providing a reliable experiment basis for the preventing of ankle injury.Methods A total of 18 volunteers participated in this study.The experimental group included 9 male with experience of parachute landing,while the other 9 male without experience of parachute landing were assigned to the control group.Each subject was instructed to jump off a platform with a height of 30 cm and 60 cm and land on a hard surface in a half-squat posture.The dynamic landing process was recorded with a high speed camera and the biomechanical data was collected and analyzed,including perpendicular ground reaction force,angular displacement,velocity and acting time.Results From 30 cm's height,the ankle angular displacement of the control group was significantly larger than the experimental group (25.73°± 8.13° vs 20.05°± 12.27°,P < 0.05).The perpendicular ground reaction force of the control group was significantly smaller than the experimental group (3 372.4±748.6 N vs 5 181.5±1 726.2 N,P < 0.05).The acting time of the control group was significantly longer than the ex perimental group (0.049±0.015 s vs 0.012±0.004 s,P < 0.05).The buffer time of the control group was significantly shorter than the experimental group (1.397±0.746 s vs 1.737±0.451 s,P < 0.05).From 60 cm's height,the ankle angular velocity of the control group was significantly higher than the experimental group (25.45± 15.01 °/s vs 16.51 ±4.18 °/s,P < 0.05).The perpendicular ground reaction force of the control group was significantly smaller than the experimental group (4 616.0±1 124.7 N vs 7 119.5±2 307.4 N,P < 0.05).The acting time of the control group was significantly longer than the experimental group (0.048±0.013 s vs 0.015±0.006 s,P < 0.05).The buffer time of the control group was significantly shorter than the experimental group (0.922±0.347 s vs 1.617±0.547 s,P < 0.05).Conclusion Jumping from different heights,the experinental group was larger in perpendicular ground reaction force but smaller in the angular velocity and displacement than the control group.There was a shorter acting time and a longer buffer time in the experimental group than the control group.
10.Application of recombinant human growth hormone combined with early enteral nutrition in severely burned patients
Zuhuang WU ; Bing SON ; Guoyang CHEN ; Yanwu ZHUO
Chinese Journal of Primary Medicine and Pharmacy 2010;17(17):2311-2313
Objective To summarize the therapeutic effect of recombinant human growth hormone and early enteral nutrition in severely burned patients. Methods 35 cases were randomly divided into group EN and group PN.Group EN was treated with early enteral nutrition(EEN) and recombiant human growth hormone(rhGH) ,while group PN was given parenteral nutrition. The plasma levels of pre-albumin (PA), C-reactive protein (CRP) , tunmor necrosis factor α(TNFα) ,fasting blood glucose(FBG) were measured after burn injury. Comparision was made in length of hospital stay, wound healing time,incidence of burn sepsis, gastrointestinal stress ulcer bleeding and enteral nutrition intoleranc. Results Incidence of burn sepsis,gastrointestinal stress ulcer bleeding and enteral nutrition intoleranc in group EN were significantly lower than those of group PN(P < 0.05). The serum levels of CRP、TNFα、FBG in group EN were significantly lower than those of group PN (P < 0.05). The serum PA was decreased at all times in two periods and was markedly decreased in group EN(P <0.05). Length of hospital stay and the healing time of donor site, deep partial thickness burn wound, skin transplantion area were significantly shortened in group EN compared with group PN. Conclusion Early enteral nutrition was beneficial for the improving of nutrition state of the burn patients,and the reduction of the incidence of burn sepsis and the hospitalization time.

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