1.Therapeutic Effect of Vanadium in Alloxan Diabetic Mice
Fei YEI ; Haifan LIU ; Mingzhi XIE
Chinese Pharmacological Bulletin 1987;0(03):-
Vanadium (V) is one of the trace elements It exists in many oxidation states which exhibit different biological activities. Our results in alloxan diabetic mice suggest that NaVO3 and VOSO4 can decrease blood glucose level. The hy-poglycemic effest of VOSO4 was found to be weaker than that of NaVO3, the toxicity ofVOSO4 is also lower. However VOSO4; showed no effect in normal mice. The mechanism of the hypoglycemic effect of VOSO4 appears to be similar to that of insulin.
2.THE THERAPEUTIC EFFECT OF WEL-LUO POWDER ON EXPERIMENTAL OBESITY AND DIABETIC MICE
Mingzhi XIE ; Guoxian SHAO ; Haifan LIU ; Yanxiong LIU
Acta Nutrimenta Sinica 1956;0(01):-
The therapeutic effect of a natural foodstuffs--Wei-Luo powder (WLP)composed of high protein, various essential amino acids, vitamins (B1, B2 and E), and trace elements (Fe, Zn, etc), on both monosodium glutamate (MSG) induced obese mice and alloxan diabetic mice were tested. After 16 days of feeding with WLP diet, the body weight and accumulation of fat in liver and adipose tissue (Lee index) in MSG mice were decreased significantly, WLP diet also caused significant reduction in carbohydrate tolerance of mice as compared with wheat diet. In addition, improvement of hyperglycemia and diabetic symptoms (polydipsia, polyuria and hyperphagia) in alloxan diabetic mice were observed. These results suggest that WLP diet may be beneficial for obesity and diabetic patients.
3.Study on the feasibility and safety of measles vaccination in women of childbearing age
Chan XIA ; Zhengwang WEN ; Qi LIU ; Lei DONG ; Haifan QIU ; Yiping CHEN
Chinese Journal of Infectious Diseases 2016;34(4):227-231
Objective To study the feasibility and safety of measles vaccination in women of childbearing age and to understand the influence of measles vaccination on the fetal transmission measles antibody level of the infants.Methods From January 1,2012 to December 31,2012,600 women of childbearing age were included in this study.The measles IgG antibody was detected,then all participants were randomized according to the IgG level.The measles IgG antibody of participants in non-vaccinated group was detected predelivery in hospital by enzyme linked-immunosorbent assay (ELISA).That of participants in vaccinated group was detected 3 months after vaccination with measles mumps rubella combined vaccine and predelivery by ELISA.And measles nuclear protein fragment gene (measles virus nucleoprotein,MVN) in the blood was detected by reverse transcription polymerase chain reaction (RT PCR).A total of 368 participants gave birth within 2 years after vaccination,and 357 infants 8 months after birth were healthy.In non-vaccinated group,8 infants were diagnosed with measles within 8 months after birth.Finally,349 8-month infants were enrolled in the study,including 52 whose mothers in high antibody without vaccination group,65 whose mothers in high antibody with vaccination group,110 whose mothers in low antibody without vaccination group and 122 whose mothers in low antibody with vaccination group.The measles IgG antibody levels in the blood of all the 8-month infants were detected.Data were analyzed using t test,one-way ANOVA (Newman-Keuls was used for comparison between groups) and Pearson analysis.Results The measles IgG antibody level of expectant women in high antibody without vaccination group was significantly lower compared to that in high antibody with vaccination group ([268.5±74.9] IU/mL vs [578.3t208.1] IU/mL,Q=15.57,P<0.01).That in low antibody without vaccination group was also significantly lower than low antibody with vaccination group ([169.4+42.3] IU/mL vs [584.7+195.8] IU/mL,Q=29.54,P<0.01).The results of MVN RT-PCR after 3 months of vaccination showed no positive bands in all blood samples.Two of the expectant women in high antibody without vaccination group and one in low antibody without vaccination group were positive for MVN bands.Among 8-month infants,the levels of antibody in high antibody without vaccination group and high antibody with vaccination group were (106.3 ± 36.8) IU/mL and (291.8±86.5) IU/mL,respectively,which was statistically different (t=23.33,P<0.01).Those in low antibody without vaccination group and low antibody with vaccination group were (87.1 ± 26.4) IU/mL and (274.0±72.5) IU/mL,respectively,which was statistically different (t =33.27,P<0.01).The measles antibody level of expectant women was positively correlated with their 8-month infants (r=0.652,P<0.01).All 8 infants who were diagnosed with measles were delivered by women without vaccination,and the measles infection rate of infants was significantly different between women with and without vaccination (P=0.002).Condusion It is feasible for women of childbearing age to receive measles vaccination,which can increase the measles IgG antibody level of both expectant women and their infants.
4.THE THERAPEUTIC EFFECTS OF A HIGH FIBRE DIET ON THE EXPERIMENTAL DIABETES AND CLINICAL DIABETES MELLITUS
Mingzhi XIE ; Guoxian SHAO ; Zhufang SHEN ; Yi HUA ; Qiming CHEN ; Haifan LIU
Acta Nutrimenta Sinica 1956;0(02):-
The therapeutic effects of a high fibre diet composed of naturally high fibre foodstuffs containing protein, various essential amino acids and trace elements (Zn, Cr etc.) on diabetics were tested both experimentally and clinically. The high fibre diet or steamed bread (for control) with identical carbohydrate content was taken by normal mice, alloxan diabetic mice and nine healthy volunteers. The high fibre diet caused significant reduction in the blood glucose area (carbohydrate tolerance) as compared with the steamed bread. In another study, eighteen non-insulin dependent diabetic patients (10 females, 8 males) were administered the high fibre diet for 34 days, the average fasting and 2 h postprandial blood glucose level were significantly lower than those before the test, but no difference was observed in blood electrolytes. This implied that no malabsorption occurred during the testing period. At the same time, 16 of the 17 overweight patients were found to have a reduction of weights. In addition, improvement of symptoms (poly-dipsia polyuria, constipation etc) both in diabetic animals and in patients was observed. This study indicates that the high fibre diet is benificial for diabetic patients.
5.Optimization of the azomethane oxide and dextran sodium sulfate model of colitis-associated colon cancer and changes in the intestinal microbiota
Dunfang WANG ; Lin ZHU ; Xue FENG ; Caijuan ZHANG ; Haifan LIU ; Yaqing LIU ; Bin LIU ; Li LIU ; Weipeng YANG
Acta Laboratorium Animalis Scientia Sinica 2024;32(2):151-160
Objective To optimize the method of combining azomethane oxide(AOM)and dextran sodium sulfate(DSS)to create a colitis-associated colon cancer(CAC)model,and to explore the pathogenesis of the intestinal flora in CAC.Methods Model groups A and B were established by one and two injections of AOM,respectively,combined with free drinking of DSS,and a normal control group was injected intraperitoneally with normal saline combined with purified water(n=10 mice per group).The better modeling scheme was selected by comprehensive evaluation of the disease activity index score,colon length,tumor rate,and mortality.Serum levels of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),and tumor markers CA199,CEA,and CA724 were detected by enzyme-linked immunosorbent assay.Colon lesions were evaluated by hematoxylin and eosin(HE)staining.Changes in the intestinal microbiota in CAC mice were detected by 16S rDNA high-throughput gene sequencing analysis of mouse feces.Results Both single and enhanced AOM injections combined with DSS induced CAC mice;however,colon growths were larger,more closely arranged,and their morphological size was more consistent in group B compared with group A,with a tumor-formation rate of 100%.IL-6 levels were increased in the model group compared with the normal group(P<0.05).TNF-α levels were increased in the model group compared with the normal group(P>0.05).The CA199 and CEA levels were also significantly increased(P<0.05),but CA724 levels were not.Infiltration of inflammatory cells in the colon detected by HE pathology was accompanied by high-grade intraepithelial tumor-like changes on the surface of the lumen.The diversity and abundance of intestinal bacteria were decreased in CAC mice compared with normal mice:phyla Verrucomicrobiota and Actinobacteriota were significantly increased(P<0.05),Bacteroidota and Campilobacterota were significantly decreased(P<0.05).Akkermansia,Prevotellaceae,Ruminococcus,and Bifidobacterium were significantly increased(P<0.05),and Roseburia,Rikenellaceae_RC9_gut_group,Anaeroplasma,and Muribaculaceae were significantly decreased(P<0.05).Conclusions Two injections of AOM combined with 1.5%(1.5 g/100 mL)DSS induced CAC model mice with a high colon-tumorigenesis rate,uniform tumor morphology,and low mortality,and may thus be the preferred modeling scheme for pharmacodynamic experiments.Disorders or dysfunction of the intestinal flora may lead to increased permeability,loss of intestinal mucosal barrier function,and the release of enterogenic endotoxins,Resultsing in a sustained inflammatory response,as an indirect or direct cause of CAC pathogenesis.
6.Effect of endomorphin 2 on expression of μ⁃opioid receptors in the dorsal root ganglion in rats with neuropathic pain
Yushan Xie ; Haifan Liu ; Tao Sun ; Faping Wan
Acta Universitatis Medicinalis Anhui 2023;58(11):1833-1840
Objective :
To explore the expression of the μ⁃opioid receptor ( MOR) and the effects of intracellular
vesicle transport on the MOR expression during endomorphin 2 ( EM2) analgesia.
Methods :
Adult male SD rats were randomly divided into 3 groups : control (naive) group , neuropathic pain group and drug group. Spared nerve injury (SNI) induced neuropathic pain rats were employed as the pain model. The drug group rats were the SNI pain ones intrathecally injected with EM2. The methods of immunofluorescence single staining and Western blot were used to detect the expression of MOR total protein , phosphorylated protein and Rab7 protein. Immunofluorescence double staining was used to detect the expression of MOR/Rab7 and MOR/LAMP1 co⁃labeled immunoreactivity.
Results :
Compared with the control group , the expression of total MOR protein and phosphorylated protein in the dorsal root ganglion (DRG) of the SNI pain rats decreased (P < 0. 05) , and the expression of Rab7 significantly increased (P < 0. 05) . The expression of MOR/Rab7 co⁃labeled immunoreactivity in Rab7 and MOR immunoreactive ( Ⅳir) products and MOR/LAMP1 co⁃labeled immunoreactivity in MOR and LAMP1 ⁃ir products both increased (P < 0. 05) . Multiple intrathecal injection of EM2 significantly increased paw withdrawal threshold in the SNI neuropathic pain rats (P < 0. 01) , the expression of MOR protein and phosphorylated protein in DRG was increased (P < 0. 05) , while the expression of Rab7 decreased (P < 0. 05) . Compared with the control group , the expression of MOR/Rab7 positive products in Rab7 and MOR positive ones decreased , and the expression of MOR/LAMP1 positive products in LAMP1 and MOR positive markers decreased ( P < 0. 05 ) .
Conclusion
In the process of analgesia , EM2 inhibits the expression of Rab7 in the DRG of SNI neuropathic pain rats , reduces the transport of MOR to lysosomes , and promotes the re⁃sensitization of MOR.
7.Sishenwan Combined with Tongxie Yaofang Treats Ulcerative Colitis with Spleen-kidney Yang Deficiency and Liver Depression
Yaqing LIU ; Haifan LIU ; Bin LIU ; Xue FENG ; Caijuan ZHANG ; Dunfang WANG ; Lin ZHU ; Weipeng YANG
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(7):40-48
ObjectiveTo induce the rat model of ulcerative colitis (UC) with spleen-kidney Yang deficiency and liver depression, and explore the efficacy and mechanism of Sishenwan combined with Tongxie Yaofang (SSW&TXYF) based on the therapeutic principles of tonifying spleen, soothing liver, warming kidney, and astringing intestine. MethodSixty male SD rats were randomized into normal, model, mesalazine, and high-, medium-, and low-dose SSW&TXYF groups. The rats in other groups except the normal group were administrated with Sennae Folium decoction and hydrocortisone and received tail clamping for 14 days. On day 14, rats received enema with TNBS-ethanol solution to induce UC. The rats were administrated with corresponding drugs from day 15 of modeling, and the body weight and mental state were observed and recorded. The sucrose preference test was performed from day 25. On day 28, the rectal temperature was measured, and the rats were administrated with 3% D-xylose solution at a dose of 10 mL·kg-1 by gavage. Blood was sampled 1 h later, from which the serum was collected for measurement of the D-xylose content. The serum, hippocampus, and colorectum samples of rats were collected on day 29. The levels of gastrin (GAS), adrenocorticotropic hormone (ACTH), corticosterone (CORT), cyclic adenosine monophosphate (cAMP), cyclic guanosine monophosphate (cGMP), interleukin (IL)-4, tumor necrosis factor (TNF)-α, and interferon (IFN)-γ in the serum and 5-hydroxytryptamine (5-HT) in the hippocampus were determined by enzyme-linked immunosorbent assay. Hematoxylin-eosin staining was employed to reveal the colonic lesions. The mRNA and protein levels of p38 mitogen-activated protein kinase (MAPK), extracellular signal-regulated kinase (ERK), and c-Jun N-terminal kinase (JNK) in the colon tissue were determined by Real-time PCR and Western blot, respectively. ResultCompared with the normal group, the model group showed decreased body weight, anal temperature, and D-xylose content in the serum and increased GAS content (P<0.01). The modeling led to cAMP/cGMP unbalance and decreased the ACTH and CORT content in the serum (P<0.01), the preference for sucrose water, and the 5-HT content in the hippocampus (P<0.01). Moreover, it shortened the colorectal length and caused massive infiltration of inflammatory cells and severe structural damage in the colon tissue. High, medium, and low doses of SSW&TXYF improved above indicators (P<0.05, P<0.01), reduced inflammatory infiltration, and repaired the pathological damage of the tissue. Compared with the normal group, the model group showed lowered IL-4 level (P<0.01) and elevated TNF-α and IFN-γ levels (P<0.05, P<0.01) in the serum, as well as up-regulated expression of p38 MAPK, ERK, and JNK (P<0.05, P<0.01). Compared with the model group, SSW&TXYF elevated the IL-4 level (P<0.01), lowered the TNF-α and IFN-γ levels (P<0.05, P<0.01), and down-regulated the mRNA and protein levels of p38 MAPK, ERK, and JNK (P<0.05, P<0.01). ConclusionA rat model of UC with spleen-kidney Yang deficiency and liver depression was successfully established. SSW&TXYF can significantly mitigate this syndrome by reducing the inflammatory response in the colon and inhibiting the MAPK pathway.
8.Intervention Mechanism of Huangqintang on Intestinal Inflammation and Proliferation in Colitis-associated Colon Cancer
Lin ZHU ; Dunfang WANG ; Xue FENG ; Caijuan ZHANG ; Haifan LIU ; Yaqing LIU ; Bin LIU ; Li LIU ; Weipeng YANG
Chinese Journal of Experimental Traditional Medical Formulae 2023;29(22):1-10
ObjectiveTo investigate the efficacy of Huangqintang on mouse models of colitis-associated colon cancer (CAC) and explore the mechanism of Huangqintang in regulating immune function and inflammatory response, inhibiting abnormal cell proliferation, and delaying or inhibiting CAC formation in CAC. MethodC57BL/6J mice were randomly divided into a normal group, model group, mesalazine group, and high- and low-dose Huangqintang groups according to body weight, with 12 mice in each group. Except for the normal group, the rest of the mice were given two intraperitoneal injections of 10 mg·kg-1 azomethane (AOM) and allowed to drink 1.5% dextran sodium sulfate (DSS) freely for seven days and water normally for two weeks. Then, two cycles of ''DSS-drinking water'' were repeated. During the administration of DSS, mice in the normal group and model group were given gavage in equal doses of pure water. Mice in the mesalazine group were given 150 mg·kg-1·d-1 mesalamine suspension for gavage, and mice in the high- and low-dose Huangqintang groups were given 18 and 9 g·kg-1·d-1 Huangqintang for gavage, respectively. Each group was given one dose daily until the end of three cycles. After the intervention, the body weight, colon length, and number of colon tumors in each group were measured, and disease activity index (DAI) scores were performed. The serum contents of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), interleukin-4 (IL-4), interleukin-10 (IL-10), and gastrointestinal tumor marker carbohydrate antigen-199 (CA199) were detected by enzyme linked immunosorbent assay (ELISA). The colonic lesions were observed by hematoxylin-eosin (HE) staining. The expression of proliferative cell-associated antigen (Ki67) was observed by immunohistochemistry. The expression of T lymphocyte subsets (CD3+, CD4+, CD8+, and CD49b+) in mouse plasma was detected by flow cytometry. Fluorescein isothiocyanate-D (FITC-D) content in mouse serum was detected by fluorescent labeling method. The Western blot method was used to detect the expression of Cyclin D1, cyclin-dependent kinase 2 (CDK2), cyclin-dependent kinase 4 (CDK4), and tightly junction-related Occludin and Claudin-1. ResultCompared with the normal group, the body weight of mice in the model group decreased. DAI score increased significantly, and the colon became shorter. Pro-inflammatory factors such as IL-6, TNF-α, and IL-1β increased, and IL-6 and TNF-α were significantly increased (P<0.05). The inflammatory factor IL-4 (P<0.05) and IL-10 were significantly reduced, and the tumor marker CA199 was significantly increased (P<0.01). HE staining showed that colon lesions, intestinal mucosal epithelial defects with a large number of inflammatory infiltrates, serious crypt structure damage, and glandular arrangement disorder were observed in the model group. Ki67 positive granules were expressed in large areas of colonic tissue. The serum CD4+ and CD4+/CD8+ of mice in the model group decreased significantly (P<0.05), and CD8+ increased significantly (P<0.05). The plasma content of FITC-D in the model group was significantly increased (P<0.05), and the expression of Cyclin D1, CDK2, and CDK4 proteins in colon tissue was significantly increased (P<0.05, P<0.01). In addition, the expression of Occludin and Claudin-1 was significantly decreased. Compared with the model group, the body weight of mice in the mesalazine group and the high- and low-dose Huangqintang groups increased. DAI score decreased, and the colon became longer. IL-6, TNF-α, and IL-1β expression decreased (P<0.05, P<0.01), but there was no significant change in IL-4 and IL-10. The content of CA199 was significantly reduced (P<0.05), and the colomatoid lesions and inflammatory infiltrates were reduced in the mesalazine group and the Huangqintang group. The crypt structure damage was lighter, and the positive expression of Ki67 was reduced. CD4+, CD4+/CD8+, and CD49b+ increased, and the difference was not statistically significant. FITC-D content decreased (P<0.05). The expression of Cyclin D1, CDK2, and CDK4 decreased (P<0.05, P<0.01), and Claudin-1 and Occludin protein expression increased in the high-dose Huangqintang group (P<0.05). ConclusionHuangqintang has a certain delay and inhibitory effect on AOM/DSS-induced inflammatory cancer transformation, and its mechanism of action may be related to regulating immune function and inflammatory response, inhibiting the release of pro-inflammatory factors, repairing damaged intestinal barriers, inhibiting abnormal proliferation of colon cells, and intervening in the formation and development of CAC colon tumors.
9.Establishment of Fingerprint and Spectrum-effect Relationship Study on Anti-oxidantion Activity of Anemarrhena asphodeloides and Its Different Processed Products
Yuxin JIA ; Haifan LIU ; Mingdong SI ; Xinrui LI ; Junna SONG ; Yuguang ZHENG ; Donglai MA
China Pharmacy 2020;31(22):2706-2712
OBJECTIVE:To establish the fingerprint of ethanol extract and acetone extract from Anemarrhena asphodeloides and its different processed products ,and to investigate the spectrum-effect relationship between the fingerprint and the antioxidant activity. METHODS :HPLC method and HPLC-ELSD method were adopted. The determination was performed on Thermo BDS Hypersil C 18 column with mobile phase consisted of acetonitrile- 0.2% acetic acid at the flow rate of 1.0 mL/min. The column temperature was 30 ℃,and the detection wavelength was set at 258 nm. The sample size was 10 μL. The determination was performed on XDB-C 18 columnwith mobile phase consisted of acetonitrile-0.1% acetic acid (gradient elution )at the flow rate of 0.9 mL/min. The column temperature was 30 ℃ . The temperature of atomizer was 40 ℃ and the flow rare of N 2 was 1.6 mL/min. The sample size was 10 μL. Using mangiferin and timosaponin B Ⅱ as reference ,Fingerprint Similarity Eva- com luation System of TCM Chromatogram (2004A edition )was adopted to draw the fingerprint of ethanol extract and acetoneextract from 20 batches of A. asphodeloides and its different processed products to confirm common peaks. Using scave nging rate of 1,1-diphenyl-2-trinitrophenylhydrazine(DPPH)radical as index,antioxidant activities of ethanol extract and acetone extract from 20 batches of A. asphodeloides and its processed products were investigated. Using scavenging rate of DPPH radical as dependent variable ,common peak area as independent variable ,PLSR was used to analyze the spectrum-effect relationship of ethanol extract and acetone extract from A. asphodeloides with antioxidantion activity. RESULTS :Eight peaks (M1-M8)were identified in the fingerprints of ethanol extracts from 20 batches of processed A. asphodeloides . Mangiferin (chromatogram peak M 7)was identified with similarity of 0.389-1.000;seven comon peaks (S1-S7)and timosaponin B Ⅱ(peak S 5)were identified in the fingerprint of acetone extract ,and the similarity was 0.044-0.999. DPPH radical scavenging rate of ethanol extract from 20 batches of A. asphodeloides and its processed products was 21.23%- 81.39%,and A. asphodeloides was significantly lower than salt-processed A. asphodeloides with salt wine-processed A. asphodeloides (P<0.001);and that of acetone extract was 49.73%-83.78%,and A. asphodeloides was significantly higher than stir-baked A. asphodeloides with salt ,wine or fire (P<0.001). The standardized regression coefficients of peaks M 2-M7 in the spectrum of ethanol extract from A. asphodeloides were all greater than 0,which was positively correlated with antioxidant activity. Only the variable importance projection (VIP)value of peak M 7 was greater than 1,which had an important contribution. The standardized regression coefficients of peaks S 4-S7 in the acetone extract spectrum of A. asphodeloides were greater than 0,and were positively correlated with antioxidant activity. The order of VIP values was peak S 5>S6>S4,and the VIP values were all greater than 1. CONCLUSIONS:The fingerprint of the different processed products A. asphodeloides and its antioxidant activity spectral effect relationship were successfully established ;mangiferin(peak M 7)may be the main antioxidant substance of ethanol extract from A. asphodeloides . Timosaponin B Ⅱ(peak S 5),peak S 6 and peak S 4 may be the main antioxidant substance in acetone extract from A. asphodeloides .