1.Promotion effect of chemokine CCL19-induced macrophage M1 polarization on chronic pancreatitis in mice and its mechanism
Lianzhi CUI ; Xiaowei ZHANG ; Yue ZHAI ; Yue PAN ; Xiuyan YU ; Hua ZHU
Journal of Jilin University(Medicine Edition) 2024;50(6):1587-1596
Objective:To discuss the promotion effect of chemokine C-C motif ligand 19(CCL19)induced macrophage M1 polarization on chronic pancreatitis of the mice,and to clarify its related mechanism.Methods:Ten male C57BL/6N mice were selected,and the pancreatic acinar cells and peritoneal macrophages were extracted from these mice to construct the macrophage-acinar cell co-culture system.The co-culture system cells were divided into control group,model group,and small interfering RNA CCL19(si-CCL19)group.The morphology of the acinar cells in various groups were observed under microscope.Forty mice were randomly selected and divided into normal group and chronic pancreatitis group,and there were 20 mice in each group.HE staining was used to observe the pathomorphology of pancreatic tissue of the mice in two groups;immunofluorescence staining was used to observe the expressions of cytokeratin 19(CK19),amylase,M1 macrophage-related markers inducible nitric oxide synthase(iNOS),and F4/80 in pancreatic tissue of the mice in two groups and morphology of follicular cells and the expressions of CK19,amylase in the co-culture system cells in various groups;enzyme-linked immunosorbent assay(ELISA)was used to detect the levels of tumor necrosis factor-α(TNF-α),interleukin(IL)-6,and IL-1β in serum of the mice in two groups and in the co-culture system cells in various groups;immunohistochemistry was used to observe the expression of CCL19 protein in pancreatic tissue of the mice in two groups;Western blotting method was used to detect the expression levels of CCL19 protein and two nuclear factor-κB(NF-κB)signaling pathway-related proteins P65,phosphorylate P65(p-P65),kappa B inhibitor of kinase α/β(IKKα/β),phosphorylated IKKα/β(p-IKKα/β),IkBα,phosphorylated IκBα(p-IκBα)in pancreatic tissue of the mice in two groups and in the co-culture system cells in various groups.Results:The HE staining results showed that the acinar cells in pancreatic tissue of the mice in normal group were tightly arranged;compared with normal group,the acinar cells of the mice in chronic pancreatitis group showed obvious vacuolation and acinar cell ductal metaplasia,indicating successful preparation of the mouse pancreatitis model.The immunofluorescence staining results showed that compared with control group,the acinar cells in model group exhibited severe vacuolation,the CK19 expression was significantly increased,and the amylase expression was significantly decreased;compared with model group,the acinar cell ductal metaplasia in si-CCL19 group was decreased,the CK19 expression was significantly decreased,and the amylase expression was significantly increased;compared with normal group,the expression of amylase in pancreatic tissue of the mice in chronic pancreatitis group was significantly decreased,while the expressions of CK19 and M1 macrophage markers iNOS and F4/80 were significantly increased.The ELISA results showed that compared with normal group,the serum levels of TNF-α,IL-6,and IL-1β of the mice in chronic pancreatitis group were significantly increased(P<0.05);compared with control group,the levels of TNF-α,IL-6,and IL-1β in the cells in model group were significantly increased(P<0.05);compared with model group,the levels of TNF-α,IL-6,and IL-1β in the cells in si-CCL19 group were significantly decreased(P<0.05).The immunohistochemistry results showed that compared with normal group,the expression of CCL19 protein in pancreatic tissue of the mice in chronic pancreatitis group was significantly increased.The Western blotting results showed that compared with normal group,the expression levels of CCL19 protein and NF-κB signaling pathway-related proteins p-P65,p-IKKα/β,and p-IκBα in pancreatic tissue of the mice in chronic pancreatitis group were significantly increased(P<0.05);compared with control group,the expression levels of CCL19,p-IKKα/β,p-P65,and p-IκBα proteins in the cells in model group were significantly increased(P<0.05);compared with model group,the expression levels of CCL19,p-IKKα/β,p-P65,and p-IκBα proteins in the cells in si-CCL19 group were decreased(P<0.05).Conclusion:CCL19 promotes the macrophage M1 polarization through the NF-κB signaling pathway,induces the formation of inflammatory microenvironment,and promotes the occurrence and development of pancreatitis.
2.GB7 acetate,a galbulimima alkaloid from Galbulimima belgraveana,possesses anticancer effects in colorectal cancer cells
Ziyin LI ; Lianzhi MAO ; Bin YU ; Huahuan LIU ; Qiuyu ZHANG ; Zhongbo BIAN ; Xudong ZHANG ; Wenzhen LIAO ; Suxia SUN
Journal of Pharmaceutical Analysis 2022;12(2):339-349
GB7 acetate is a galbulimima alkaloid obtained from Galbulimima belgraveana.However,information regarding its structure,biological activities,and related mechanisms is not entirely available.A series of spectroscopic analyses,structural degradation,interconversion,and crystallography were performed to identify the structure of GB7 acetate.The MTT assay was applied to measure cell proliferation on human colorectal cancer HCT 116 cells.The expressions of the related proteins were measured by Western blotting.Transmission electron microscopy(TEM),acridine orange(AO)and monodansylcadaverine(MDC)staining were used to detect the presence of autophagic vesicles and autolysosomes.A transwell assay was performed to demonstrate metastatic capabilities.Oxygen consumption rate(OCR)and extracellular acidification rate(ECAR)assays were performed to determine the mitochondrial oxidative phosphorylation(OXPHOS)and glycolysis activity of HCT 116 cells.The data showed that GB7 acetate suppressed the proliferation and colony-forming ability of HCT 116 cells.Pretreatment with GB7 acetate significantly induced the formation of autophagic vesicles and autolysosomes.GB7 acetate upregulated the expressions of LC3 and Thr172 phosphorylated adenosine 5'-monophosphate(AMP)-activated pro-tein kinase α(p-AMPKα),which are key elements of autophagy.In addition,GB7 acetate suppressed the metastatic capabilities of HCT 116 cells.Additionally,the production of matrix metallo-proteinase-2(MMP-2)and MMP-9 was reduced,whereas the expression of E-cadherin(E-cad)was upregulated.Furthermore,GB7 acetate significantly reduced mitochondrial OXPHOS and glycolysis.In conclusion,the structure of the novel Galbulimima alkaloid GB7 acetate was identified.GB7 acetate was shown to have anti-proliferative,pro-autophagic,anti-metastatic,and anti-metabolite capabilities in HCT 116 cells.This study might provide new insights into cancer treatment efficacy and cancer chemoprevention.
3.Relationship between serum fibroblast growth factor 21/23 level, trauma severity and prognosis in middle-aged and older adult patients with traumatic fracture
Xiaoshan WU ; Jian LI ; Jie JI ; Dandan LI ; Lianzhi ZHANG ; Xiaobin CAI
Chinese Journal of Primary Medicine and Pharmacy 2021;28(12):1814-1819
Objective:To investigate the relationship between serum fibroblast growth factor 21/23 level, trauma severity and prognosis in middle-aged and older adult patients with traumatic fracture.Methods:A total of 126 middle-aged and older adult patients with traumatic facture who received treatment in the Second People's Hospital of Lishui, China between June 2017 and June 2019 were included in the study group. Fifty healthy controls who concurrently received physical examination in the Second People's Hospital of Lishui were included in the control group. The study group was divided into five subgroups according to relevant criteria: mild, moderate, severe, poor prognosis and good prognosis. The levels of C-reactive protein (CRP), procalcitonin (PCT), FGF21 and FGF23 were measured.Results:On admission, serum CRP, PCT, FGF23, FGF2 levels in the study group were (19.18 ± 5.66) mg/L, (0.71 ± 0.20) μg/L, (79.75 ± 18.62)μg/L,(52.10 ± 16.34) μg/L, respectively, and they were significantly higher than those in the control group [ (7.60 ± 2.61) mg/L, (0.30 ± 0.11) μg/L, (40.18 ± 10.33) μg/L, (30.11 ± 10.19) μg/L, t = 18.888, 17.750, 18.336, 11.032, all P < 0.001). On admission, serum CRP, PCT, FGF23, FGF2 levels in the study group were (19.18 ± 5.66) mg/L, (0.71 ± 0.20) μg/L, (79.75 ± 18.62) μg/L, (52.10 ± 16.34) μg/L, respectively, and they were significantly increased at 1 day [(21.59 ± 4.53) mg/L, (0.79 ± 0.22) μg/L, (83.85 ± 19.07) μg/L, (55.18 ± 16.55) μg/L, t = 3.72, 3.29, 1.56, 1.56, P < 0.05, P < 0.05, P = 0.122, P = 0.122] and 3 days after surgery [(23.15 ± 3.16) mg/L, (0.80 ± 0.24) μg/L, (88.11 ± 19.80) μg/L, (59.70 ± 16.07) μg/L, t = 6.65, 3.12, 3.59, 3.77, all P < 0.05] , and significantly decreased at 7 days after surgery [(14.35 ± 4.02) mg/L, (0.52 ± 0.16) μg/L, (50.06 ± 15.50) μg/L, (32.18 ± 12.52) μg/L, t = 8.31, 8.58, 13.77, 11.11, all P < 0.001]. On admission, there were significant differences in serum CRP, PCT, FGF23, FGF21 levels between mild, moderate and severe groups ( F = 25.087, 15.851, 15.831 and 12.645, all P < 0.001). On admission, serum CRP, PCT, FGF23, FGF21 levels in the poor prognosis group were significantly higher than those in the good prognosis group ( t = 5.757, 4.984, 3.189 and 4.006, all P < 0.001). The receiver operating characteristic curve (ROC) analysis results showed that serum CRP, PCT, FGF23, FGF21 levels in patients with traumatic fracture on admission had a certain value in the prediction of poor prognosis. Combined detection of these four indexes had the highest value, with AUC (0.95 CI) of 0.877 (0.783-0.982). Conclusion:Serum FGF21 and FGF23 levels have a certain value in the prediction of severity and prognosis of traumatic fracture in middle-aged and older adult patients.
4.Preparation of warangalone-loaded liposomes and its inhibitory effect on breast cancer cells.
Lianzhi MAO ; Huiping LIU ; Huahuan LIU ; Zhongbo BIAN ; Qiuyu ZHANG ; Wenzhen LIAO ; Suxia SUN
Journal of Southern Medical University 2020;40(6):876-883
OBJECTIVE:
To prepare warangalone-loaded thermosensitive liposomes (WLTSL) and evaluate its inhibitory effect on breast cancer cells .
METHODS:
MTT assay was used to assess the changes in proliferation of 3 breast cancer cell lines (MDA-MB-231, MCF7, and SKBR3) following treatment with warangalone, soy isoflavone and genistein. Colony-forming assay and wound healing assay was used to assess colony forming activity and migration of MDA-MB-231 cells treated with warangalone. The effect of warangalone on the expression of MMP2 and MMP9 in MDA-MB-231 cells was examined with Western blotting. The thermosensitive liposomes (TSL) and WLTSL were prepared using a thin film hydration method, and the morphology, size, encapsulation efficiency and stability of the prepared liposomes were characterized using transmission electron microscopy, dynamic light scattering scanning and UV spectrophotometry. MTT assay was used to examine the inhibitory effect of WLTSL on mouse breast cancer cells (4T1) .
RESULTS:
Warangalone showed stronger anti-proliferation effects than soy isoflavones and genistein in the 3 human breast cancer cell lines and significantly inhibited colony formation by MDA-MB-231 cells. Treatment with warangalone significantly inhibited migration of the breast cancer cells and down-regulated the cellular expressions of MMP2 and MMP9. The prepared TSL and WLTSL presented with a homogeneous, irregular spherical morphology, with a mean particle size of 56.23±0.61 nm, a polymer dispersity index of 0.241±0.014, a Zeta potential of -40.40±0.46 mV, and an encapsulation efficiency was 87.68±2.41%. WLTSL showed a good stability at 4 ℃ and 37 ℃ and a stronger inhibitory effect than warangalone in 4T1 cells.
CONCLUSIONS
Warangalone inhibits the proliferation, migration and invasion of breast cancer cells, and the prepared WLTSL possesses good physical properties and strong anti-breast cancer activity.
Animals
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Breast Neoplasms
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Cell Line, Tumor
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Cell Movement
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Cell Proliferation
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Humans
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Isoflavones
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Liposomes
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Mice
5.Study on the Mechanism of Celosia cristata N-butanol Extracts in the Improvement of Dysfunctional Uter-ine Bleeding of Rats
Yuanyuan GU ; Runqin ZHAO ; Lang SHI ; Yunfei ZHANG ; Cheng FENG ; Fang FANG ; Shuxiang ZHANG ; Lianzhi WANG ; Dazhong CHEN
China Pharmacy 2017;28(19):2617-2619
OBJECTIVE:To study the improvement effect of Celosia cristata n-butanol extracts on dysfunctional uterine bleed-ing of rats,and explore its mechanism. METHODS:60 pregnant SD rats were randomly divided into blank group,model group, Gongxuening capsule group (positive control,0.07 g/kg) and C. cristata n-butanol extracts high-dose,medium-dose,low-dose groups(4.32,2.16,1.08 g/kg),10 in each group. Except for the blank group,rats in other groups were intragastrically given mife-pristone and misoprostol on 7th of pregnancy for resulting incomplete abortion to induce models of dysfunctional uterine bleeding. Then rats in administration groups were intragastrically given relevant medicines,rats in blank group and model group were intra-gastrically given normal saline once every morning and evening,for 7 d. On 8th d of pregnancy,uterine bleeding amount,and thromboxane (TXA2),prostacyclin (PGI2) and tumor necrosis factor α(TNF-α) contents in serum were determined. RESULTS:Compared with blank group,uterine bleeding amount in model group was significantly increased(P<0.01),TXA2 content in se-rum was significantly reduced,PGI2 and TNF-α contents were significantly increased(P<0.01). Compared with model group,uter-ine bleeding amounts in administration groups were significantly reduced,TXA2 content in serum was significantly increased(P<0.01);PGI2 and TNF-α contents in serum in Gongxuening capsule group and C. cristata n-butanol extracts high-dose group and TNF-α content in serum in C. cristata n-butanol extracts medium-dose group were significantly reduced (P<0.01). CONCLU-SIONS:C. cristata n-butanol extracts show obvious improvement effect on incomplete drug abortion-induced dysfunctional uterine bleeding of rats,and the mechanism may be related to the regulation of TXA2/PGI2 dynamic balance and inhibition of TNF-α tran-sient secretion.
6.Study on HPLC Fingerprints of Schisandrae Chinensis Fructus from Different Producing Areas
Lianzhi WANG ; Ke FU ; Yuanyuan GU ; Lei LIU ; Shuxiang ZHANG ; Qi ZHOU
Chinese Journal of Information on Traditional Chinese Medicine 2016;23(11):103-106
Objective To establish the fingerprint analysis method of Schisandrae Chinensis Fructus by HPLC; To analyze the similarity on the fingerprints of Schisandrae Chinensis Fructus from different producing areas. Methods The chromatographic separation was performed by HPLC on a Diamonsil C18 column (250 mm×4.6 mm, 5 μm). Acetonitrile-water was used as gradient mobile phase. The flow rate was 1.0 mL/min. The column temperature was maintained at 30℃. The detection wavelength was set at 254 nm.Results The HPLC fingerprint analysis method of Schisandrae Chinensis Fructus was established. Twenty-nine common fingerprint peaks were identified. The similarities of the fingerprints of ten samples from different producing areas were above 0.95.Conclusion The method is simple and reliable, which can provide a scientific basis for quality evaluation of Schisandrae Chinensis Fructus.
7.Application Comparison of Potassium Magnesium Aspartate before and after the Intervention of Clinical Pharmacists
China Pharmacist 2015;(8):1330-1332
To explore the effect of the intervention of clinical pharmacists on the rational use of potassium magnesium aspartate . Methods:The prescriptions of outpatients in the fourth quarter of 2013 and those of emergency in the second quarter of 2014 with potassium magnesium aspartate injection were collected and analyzed. The unreasonable use rate of the drug before and after the intervention was compared. Results:The unreasonable use rate of potassium magnesium aspartate before the intervention was 35. 4%, that after the intervention was 15. 8%, and the difference was statistically significant(P<0. 01) . The difference in beyond the indica-tion, high concentration and unreasonable compatibility of the drug before and after the intervention was statistically significant ( P<0. 01 or P<0. 05) . Conclusion: The intervention of clinical pharmacists in the use of potassium magnesium aspartate is effective, which should be performed continuously.
8.A Novel Biomimetic Porphyrin Biosensor Based on Poly (3,4-ethylenedioxythiophene) and 1-Pyrenebutanoic Acid Decorated with Hematin and Its Application in Bioelectrical Catalysis of Catechol
Xuehua YU ; Jinming KONG ; Lianzhi LI ; Xueji ZHANG
Chinese Journal of Analytical Chemistry 2014;(10):1400-1407
A new biosensor was prepared based on conductive polymer poly ( 3 , 4-ethylenedioxythiophene ) ( PEDOT) and 1-pyrenebutanoic acid ( PBA) through π-π stacking, as well as hematin associated with PBA through coordinate bonds of Zr4+ and carboxyl group. Its stability and sensitivity were examined by cyclic voltammetry ( CV) , electrochemical impedance spectroscopy ( EIS) and current-time ( i-t) method. A pair of well defined and quasi-reversible redox peaks was observed when GCE/PEDOT/PBA/hematin was tested by CV in PBS without oxygen. The electron transfer rate constant was estimated to be 4. 8 s-1 . Results indicated that the PEDOT film enhanced the electron transfer process of hematin. When the i-t method was used to detect the response of biosensor to catechol with different concentrations, it displayed a linear response for the reduction of catechol in the range of 0. 5-200 μmol/L. The linear equation was i=0. 018C+0. 006 ( R=0. 9998), and the detection sensitivity was 0. 258 μA (μmol/L· cm2) with a detection limit of 0. 33 nmol/L (S/N=3). The results illustrate that the GCE/PEDOT/PBA/hematin biosensor is very sensitive and stable.
9.Clinical study of bilateral decompressive craniotomy for treating patients with double hedge severe craniocerebral injury
Chinese Journal of Postgraduates of Medicine 2012;35(17):4-7
Objective To investigate the clinical efficacy of bilateral decompressive craniotomy for treating the patients with double hedge severe craniocerebral injury (DHSCBI).Methods Sixty patients with DHSCBI were divided by random digits table method into treatment group and control group with 30 cases each.The treatment group was treated with bilateral decompressive craniotomy,while the control group was treated with traditional unilateral craniotomy.The acute brain swelling,incisional hernia,intracranial pressure of postoperative 1,3,7 d were observed and recorded.The patients were followed up for 3 months to evaluate the clinical efficacy.Results The incidence of acute brain swelling and incisional hernia in treatment group were 13.3% (4/30) and 20.0% (6/30),respectively,which were significantly lower than those in control group [56.7% (17/30) and 63.3% (19/30)] (P <0.01 ).The intracranial pressure of postoperative 1,3,7 d in treatment group [(21.34 ±3.05),(18.43 ±2.63),(15.52 ±2.21) mm Hg(1mm Hg =0.133 kPa)] were significantly lower than those in control group[ (31.21 ± 4.46),(29.13 ±4.16),(24.97 ±3.57) mm Hg] (P <0.05).The clinical efficacy in treatment group [53.3% (16/30)] was significantly higher than that in control group [23.3% (7/30)] (P < 0.05 ).The mortality rate in treatment group [16.7% (5/30)] was significantly lower than that in control group [40.0%(12/30)] (P <0.05).Conclusions Compared with traditional unilateral craniotomy,bilateral decompressive craniotomy can reduce the incidence of acute brain swelling and incisional hernia of DHSCBI patients,and have better prognosis.It is worthy of clinical application.
10.A novel capsule-style manometric system and its application on non-invasive measurement of pancolonic pressure.
Wenqiang ZHANG ; Guozheng YAN ; Lianzhi YU ; Xinqing YANG
Journal of Biomedical Engineering 2008;25(5):1079-1083
Pan-colonic motility was studied under normal conditions with a novel capsule-style system. A single use telemetry capsule embedded with one pressure sensor was ingested by subjects. It is capable of transmitting colonic pressure wirelessly greater than 130 h. Time of capsule entering segmental colon was determined by ultrasonic detection for tracing the batteries in capsule. The ultrasonic electrodes were mounted on the surface of subjects' ileocecum, navel as well as the junction of left colon and rectosigmoid colon in sequence. They were identified by abdominal X-rays with radiopaque markers. The confirming X-rays showed all telemetry capsules were detected successfully when passing through the key points in colon. A total of 613 h of colorectal recording was obtained from 20 healthy subjects. When compared with the parameters in the time of waking, the number of contractions and the area under contractions were significantly (P<0.05) decreased during sleep (21 +/- 5 vs 15 +/- 4 h(-1); 463 +/- 54 vs 342 +/- 45 mmHg x s x min(-1)). The colonic motility exhibited significant regional variation in the circadian behavior, as well as in its response to waking and meal. The clinical study proved the reliability and non-invasiveness of the system. It may represent a useful tool for the study on physiology and pathology of colonic motor disorders.
Adult
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Capsules
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Colon
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physiology
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Female
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Gastrointestinal Motility
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physiology
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Humans
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Male
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Manometry
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instrumentation
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methods
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Middle Aged
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Pressure
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Telemetry
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instrumentation
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Ultrasonics
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Young Adult

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