2.The damages of adrenal gland in rabbits with neisseria memingococcal septic shock
Kaihe DU ; Guoqing YIN ; Bei ZHONG
Chinese Journal of Pathophysiology 2001;17(6):566-568
AIM: To investigate the damages of adrenal gland in rabbits infected with neisseria meningococci(MC).METHODS: 18 rabbits were injected with MC (1×1011 organism/kg, iv, model group, n=10) or normal saline (control group, n=8).TNF-α was detected in baseline (before challenged), 60 min and 120 min after injection. 120 min after injection, TNF-α immunochemical localization of adrenal gland was detected with ABC methods by electron microscopy. RESULTS: In model group, TNF-α positive cells were located in medulla and TNF-α released into simus of medulla. The medulla damages were appeared but gland cells of cortex was normal. No damages were showed in control group. TNF-α value of plasm was elevated at 60 min and 120 min after injection compared with control group. CONCLUSION: TNF-α was released into medulla and resulted in medulla damages after MC challenging. Adrenal cortex didnt damaged in early phase of MC septic shock.
3.Inhibitory effects on Candida albicans of vagina cells transferred with antimicrobial peptide LL-37 and human defensin 5 recombinant plasmids
Fang WANG ; Bei SUN ; Hong LI ; Lirong YIN
Chinese Journal of Obstetrics and Gynecology 2012;47(3):205-211
Objective To evaluate the inhibitory effects on Candida albicans of vagina cells transferred with antimicrobial peptide LL-37 and human defensin 5 (HD5) recombinant plasmids and observe secretion of IL-8.Methods ( 1 ) The epithelial cells from female vagina were culture primarily.The pcDNA3.1 ( + )/HD5-EGFP and pcDNA3.1 ( + )/LL-37-EGFP eukaryotic recombinant plasmids were separately or jointly transferred into the fourth generation of vaginal epithelial cells.Two test groups were defined:one test group was no Candida albicans group including four subgroups which were untransferred group,HD5 group transferred with pcDNA3.1 ( + )/HD5-EGFP,LL-37 group transferred with pcDNA3.1 (+)/LL-37-EGFP,combining transferring group transferred with pcDNA3.1 ( + )/HD5-EGFP and pcDNA3.1 ( + )/LL-37-EGFP; the other test group was with Candida albicans group which the Candida albicans were coincubated with the four subgroups described above.(2) For examination of cytokines and chemokines,at 6,12,24 and 48 hours,the supernatant of every group was collected.ELISA was applied to detect the levels of LL-37,HD5 and IL-8.At each time point,the growth inhibition of Candida albicans was calculated by glucose consumption testing.Results ( 1 ) The max level of LL-37,HD5 and IL-8 reached max level after being transferred for 24 hours,then showed decreasing trend.The secretion of LL-37,HD5 and IL-8 was significant higher in combining transferring group in Candida albicans group than other groups,and the secretion level of LL-37,HD5 and IL-8 was (100.16 ±0.81 ) ng/ml,(58.50 ±2.08) μg/ml and ( 101.03 ± 1.59) pg/ml (P <0.01 ).(2) In different time point,the absorbance of each subgroup without Candida albicans declined slowly,and there were no statistically significant difference (P >0.05 ),as while as in LL-37 subgroup and HD5 subgroup with Candida albicans.In group with Candida albicans,the absorbance of combining transferring subgroup were 3.210 ± 0.010,3.150 ± 0.030,3.099 ± 0.030 and 2.970 ±0.040 at 6,12,24 and 48 hours,respectively,which was significantly higher than those in the other cells (P < 0.01 ),and the declined trend was the slowest.Conclusions The antifungal ability of vaginal epithelial cell became stronger after being transferred with LL-37 and HD5 recombinant plasmids.LL-37 and HD5 could also possess immunomodulatory activity and induce chemokine IL-8 production.
4.Prokaryotic Expression and Antifungal Activity of HumanαDefensin-5 Protein
Pingping ZHANG ; Lirong YIN ; Fang WANG ; Bei SUN ; Yan HUO
Tianjin Medical Journal 2014;(8):755-758
Objective To construct the prokaryotic expression vector for HD-5 and purify the recombinant HD-5 protein then analyze its antifungal activity. Methods The HD-5 gene was cloned by PCR, then was inserted into prokary-otic expression plasmid pQE-30Xa to construct pQE-30Xa/HD-5. After sequencing, pQE-30Xa/HD-5 was transformed in-to E.coli M15 cells. Its expression was induced by IPTG and confirmed by SDS-PAGE. The recombinant protein was purified through Ni-NTA affinity purification system. The antifungal activity was tested by disk diffusion method. Results HD-5 gene and pQE-30Xa/HD-5 vector were obtained successfully. E.coli M15 strains was used to express HD-5 fusion protein. After purification, the fusion protein was confirmed by Western blot. The disk diffusion test confirmed that the fusion pro-tein can inhibit Candida albicans. Conclusion Expression vector pQE-30Xa/HD-5 was successfully constructed. The HD-5 fusion protein was expressed in E.coli successfully, which showed a certain degree of antifungal activity.
5.Effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes in the murine model of vaginal candidiasis
Fang WANG ; Yan HUO ; Lirong YIN ; Bei SUN ; Pingping ZHANG
Chinese Journal of Obstetrics and Gynecology 2016;51(7):530-534
Objective To study the effects of antimicrobial peptide LL-37 expressed and purified from prokaryotes on candida albicans growth. Methods (1) Thirty female Kunming mice were treated with estrogen and white candida yeast suspension were poured into vagina to establish a vulvovaginal candidiasis (VVC) murine model. After successful establishing the VVC mouse model, mice were randomly sorted into test group (n=15) and control group (n=15) . Suspension(30μl, 100μg/ml)of recombinant peptide LL-37 expressed and purified in Prokaryotes was given by intravaginal administration to the test group for 5 days, while the same amount of phosphate buffered saline (PBS) was given to the control group. (2) Tweenty-four hours after treatment, the fungal burden and colony-forming unit (CFU) of vaginal fluids were evaluated. All mice were subsequently sacrificed and vaginal tissues were harvested for tissue homogenate preparation. ELISA was used to determine the levels of nterleukin-10(IL-10)and interferon-γ(IFN-γ)in the isolated vaginal tissues. Results (1) VVC mouse model was established successfully in this study. Vaginal mucosa congestion, edema, vaginal plica disappearing were obviously observed in the control group. After treatment with recombinant protein LL-37 vaginal mucosa has no obvious change in the test group. (2) Fungal burden and CFU of vaginal fluids were significantly lower in the test group [(4.8±1.0)×104 CFU/ml] than that in the control group [(8.5 ± 2.1) × 104 CFU/ml, P=0.017]. IFN-γlevel of the test group was increased [(257 ± 11) vs (197 ± 4) pg/ml, P=0.000], while the level of IL-10 was reduced [ (287 ± 15) vs (379 ± 17) pg/ml P=0.000] resulting in a the ratio of IFN-γ/IL-10 was in significantly higher in test group (0.892±0.008 vs 0.496±0.013, P=0.000). Conclusion Recombinant protein LL-37 expressed and purified from prokaryotes inhibits the growth candida albicans and improves vaginal immunity by adjusting IFN-γand IL-10 secretion in the VVC mouse model, highlighting the therapeutic potential of LL-37 for VVC.
6.Components in Antineoplastic Actinomycete Strain(N2010-37)of Bottom Mud in Mangrove
Zhongliu ZHOU ; Bei JIN ; Wenqing YIN ; Chunyan FU ; Huafen FENG
Chinese Herbal Medicines 2011;03(3):165-167
Objective To study the antitumor components from an actinomycete strain(N2010-37)of bottom mud in Zhanjiang Mangrove,South China Sea.Methods The components were isolated and purified by chromatographic techniques and recrystallization,and the structures were identified by spectral methods together with physicochemical analyses.The antitumor effects of these components were tested in vitro by MTT method.Results Three compounds were identified including two anthrones and one novel lactone.They are(3S,4R,7R,8R,9S)-3,8-dihydroxy-4,7,9-trimethyl-2,6-cyclononanediolacetone(1),2-hydroxy-l-methoxy-3-methylanthraquinone(2),and 1,6,8-thihydroxy-3-methyl-anthraquinone(3).Conclusion Compound 1 is a new compound,and compounds 1 and 3 show the favorablecytotoxic activities against human chronic granulocytic leukemia cell line K562 strain by MTT method in vitro.
7.Therapeutic Observation of Surround Electroacupuncture in Relieving Pain After Operation for Mixed Hemorrhoids
Junyi WU ; Bei CHEN ; Xinlin ZHANG ; Ping YIN ; Shifen XU
Shanghai Journal of Acupuncture and Moxibustion 2016;35(6):680-683
Objective To observe the clinical efficacy of surround electroacupuncture in relieving pain after operation for mixed hemorrhoids. Method Seventy-two patients with mixed hemorrhoids were randomized into a treatment group and a control group, 36 cases in each group. The two groups both received the corresponding interventions once in 15 min after the operation: the treatment group received electroacupuncture treatment, and the control group was only placed in a simulated environment of electroacupuncture but without receivimg any needling treatment. The pain intensity was evaluated by using Verbal Rating Scale (VRS), Wong-Baker Faces Pain Rating Scale, and Visual Analogue Scale (VAS) at 11 different time points after the treatment: once every 1 h in 8 h after the treatment, once at the first defecation, once respectively 24 h and 48 h after the treatment. The data were statistically analyzed afterwards. Result At 3 h and 4 h after the treatment and at the first defecation after the treatment, the VRS scores of the treatment group were significantly lower than that of the control group (P<0.05); respectively at 4 h, 5 h, 6 h, 7 h, and 8 h after the treatment and at the first defecation, the Wong-Baker scores of the treatment group were significantly lower than that of the control group (P<0.05); respectively at 2 h, 3 h, 4 h, 5 h, 6 h, 7 h, and 8 h after the treatment, at the first defecation, as well as at 24 h after the treatment, the VAS scores of the treatment group were significantly lower than that of the control group (P<0.05); it indicates that the pain intensity of the treatment group should be lower than that of the control group at the above time points, and the analgesic effect in the treatment group lasts a longer time, running a lower risk of losing effectiveness compared to the control group (P<0.05). Conclusion Real-time acupuncture treatment after the operation is significantly effective in relieving pain after operation for hemorrhoids.
8.Cinepazide Maleate Combined with Venlafaxine for Post-stroke Depression
Huanjie HUANG ; Bei SHAO ; Weiyong YIN ; Dalou HE
Chinese Journal of Rehabilitation Theory and Practice 2011;17(4):368-369
ObjectiveTo investigate the effect of cinepazide maleate combined with venlafaxine on post-stroke depression.Methods60 stroke patients with post-stroke depression were divided into control group (n=30) and observation group (n=30). On the basis treatment, the control group was given venlafaxine, the observation group was given venlafaxine and maleate cinepazide injection. The course was 4 weeks. The two groups were assessed with Hamilton Depression Scale(HAMD) and modified Barthel Index(MBI) before and after treatment.ResultsThe observation group improved better than the control group (P<0 05) even in HAMD and MBI 4 weeks after treatment(P<0.01).ConclusionCinepazide maleate combined with venlafaxine may be more effective on post-stroke depression to improve depressive symptoms, and the activities of daily living.
9.Analysis of intraspecific variation of Chinese Carthamus tinctorius L. using AFLP markers.
Lei ZHANG ; Bei-Bei HUANG ; Guo-Yin KAI ; Mei-Li GUO
Acta Pharmaceutica Sinica 2006;41(1):91-96
AIMTo investigate the intraspecific variation of Carthamus tinctorius L. (safflower) and establish foundation for further breeding of safflower germplasm resource and screening the quality correlation genes.
METHODSAmplified fragment length polymorphism (AFLP) was carried out to analyze genetic variation of 28 safflower populations collected in China. Unweighed pair-group method of with arithmetical averages (UPGMA) cluster analysis was used to construct a dendrogram and to estimate the genetic distances among the populations.
RESULTSAll populations could be uniquely distinguished using 12 selected primer combinations. Similarity coefficients ranged from 0.48 to 0.96 among the populations. Dendrogram revealed distinct segregation of all the cultivars into three main groups and one midst group.
CONCLUSIONLimited genetic diversity exists within the tested 28 collections at intra specific level and AFLP-based phyiogeny was not absolutely consistent with that based on morphological characters may be due to the interaction effect between genotype and environment.
Carthamus tinctorius ; genetics ; Cluster Analysis ; DNA, Plant ; analysis ; Genetic Variation ; Phylogeny ; Plants, Medicinal ; genetics ; Polymorphism, Genetic
10.Design, synthesis and in vitro activity of glycinamide-bearing compounds as DPP-IV inhibitors.
Bei HAN ; Yi HUAN ; Ziyun LIN ; Peng LI ; Zhufang SHEN ; Dali YIN ; Haihong HUANG
Acta Pharmaceutica Sinica 2010;45(11):1379-84
To research the structure-activity relationship (SAR) of glycinamide-bearing compounds that used as inhibitors of dipeptidyl peptidase IV (DPP-IV), P32/98 and compound A were chosen as the leading compounds, heterocycles containing nitrogen atom were introduced to form amide, and different residues on a-position of carbonyl were designed. The nineteen designed compounds were synthesized by a simple route and were evaluated as inhibitors of DPP-IV. All of the structures were characterized by 1H NMR and HRMS. The preliminary SAR result was obtained.