1.A Preliminary Study on Establishment of Cellular Injury Models of Blood Stasis Syndrome
Yunbo CHEN ; Qi WANG ; Shilong LAI
Journal of Guangzhou University of Traditional Chinese Medicine 1999;0(02):-
Objective To explore methods for establishinga cellu lar injury model of blood stasis syndrome (BSS). Methods Two experiments were c arried out. In the first experiment,the injured vascular endothelial cell model of BSS was established by the culture of vascular endothelial cell, and in the seco nd experiment, the vascular endothelial cell injured by the serum of BSS rabbit was cultured. Results In the first experiment, pathological features and end ocrine dysfunction in the primary cultured endothelial cell were similar to the B SS rabbit model, but the above changes did not remain in the subculture. The above changes were also found in the second experiment and were easy to be repea ted. Conclusion The injured vascular endothelial cell model of BSS establis hed b y the above methods reflects or partially reflects the structure and function of BSS rabbit model and the patients with BSS, and can be used to study the pathol ogical mechanism of BSS and therapeutic effect of blood_circulation activating a nd blood_stasis removing herbs.
2.Validation of the silencing site of marmoset B2m gene at the cellular level
Yichen DENG ; Chen ZHANG ; Zhiguang XIANG ; Yongkang TENG ; Yunbo LIU
Chinese Journal of Comparative Medicine 2017;27(5):37-41
Objective To screen and determine the effective silencing targets of β2-microglobulin(B2m)gene at the cellular level in marmoset.Methods By homology comparison of the b2m gene in human and the B2m gene in marmoset, choose homology small hairpin RNA(shRNA)sequences targeting marmoset B2m gene were designed, We choose homology small hairpin RNA(shRNA)sequences targeting designed B2m gene to make homology analysis, and insert into lentivirus-based gene silencing constructs FUGW-TDT.The vectors were transfected into HEK293T cells induced by polyethylenimine(PEI).The suppression of B2m mRNA was detected by real-time PCR.Results Two gene-silencing sequences were screened that lied in 290~310 bp and 665~685 bp of the marmoset B2m mRNA, and have statistical significance in the silencing rate:(46.54±7.91)% (P < 0.05) and(83.22±4.37)%(P < 0.0001).Conclusions Two effective silencing target sequences are screened at cellular level, which can be further used in studies on gene silencing in marmoset.
3.Effect of Shengmai capsules on MMP-3&TIMP-1 of cardiac muscle in rats with chronic congestive heart failure
Yuanjiang DENG ; Weixiong LIANG ; Weiying LIU ; Shuyi CHENG ; Yunbo CHEN
China Journal of Traditional Chinese Medicine and Pharmacy 2005;0(11):-
Objective:To research the effect of Shengmai capsules on the matrix metalloproteinase(MMPs)& tissue inhibitor of metalloproteinase(TIMPs)in chronic congestive heart failure(CHF)rats.Methods:75 SD female rats weredivided randomly into 5 groups:sham-operation group(A),CHF model group(B),CHF model treated by Shengmai capsules group(Shengmai capsule group,C),CHF model treated by Captopril group(Captopril group,D),CHF model treated by Shengmai capsule and Captopril group(shengmai capsule&captopril group,E),15 rats each group.Suprarenal abdominal artery constriction was operated to prepare CHF rat models.After 7 week treating respectively,the contents of MMP-3&TIMP-1 of cardiac muscle in rats were detected.Results:The content of cardiac MMP-3 was higher in group B than in group A(P
4.Clinical analysis of diagnosis and treatment of primary orbital tumors
Rong CHEN ; Yan LIU ; Jiangnan XIA ; Qian ZHANG ; Yunbo YANG
Chinese Journal of Postgraduates of Medicine 2014;37(33):55-57
Objective To explore the clinical manifestation,imaging features,histopathological classifications and treatment of primary orbital tumors.Methods Twenty-six cases with primary orbital tumors were retrospectively studied.Results All of 26 primary orbital tumor cases received surgical treatment.Sixteen primary orbital tumors cases were male and 10 cases were female.The mean age was 46 years (ranged from 15 to 72).The mean hospital stay was 13 d (ranged from 9 to 21).Among 26 primary orbital tumors cases,21 cases were benign tumors which included 11 cases of cavernous hemangioma,5 cases of inflammatory pseudotumor,3 cases of dermoid cyst,2 cases of venous angioma.Five cases were malignant tumors which included 4 cases of adenoid cystic carcinoma and 1 case of rhabdomyosarcoma.After operation,visual acuity improved in 9 cases,unchanged in 11 cases,decreased in 6 cases.The patients were followed up for 18-48 months (mean,25 months).There were 4 cases of malignant tumors recurrence after operation and received radical operation.While 2 patients were lost,the other 24 patients survived with tumor-free.Conclusions Surgical excision is the main and effective treatment for primary orbital tumors.To be very familiar with the imaging characteristics and local anatomy is the key for operation.Individualized treatment plan should be chosen based on clinical manifestation,imaging features and histopathological classifications.
5.Effects of drug serum in broken bushen yizhi formulas on cell model of Alzheimer disease
Yunbo CHEN ; Shilong LAI ; Jingqing HU ; Qi WANG ; Shuyi CHENG
Chinese Journal of Tissue Engineering Research 2005;9(32):250-253
BACKGROUND:Chinese herb, bushen yizhi formula protects at certain extent learning and memory in rat model of Alzheimer disease. The drug serum in this formula can alleviate neurotoxic reaction of nerve tumor cell NG 108-15 to beta-amyloid protein. In order to understand further the mechanism and compatibility of the formula, it is necessary to carry on the study on the broken formulas.OBJECTIVE:To study the effect of drug serum in subgroups of broken bushen yizhi formulas on growth and differentiation of cell model of Alzheimer disease and probe into the compatibility rule of bushen yizhi formula in view of serum pharmacology.DESIGN: Randomized controlled experiment.SETTING: DME Center of Clinical Pharmacological Institute Affiliated to Guangzhou University of Traditional Chinese Medicine.PARTICIPANTS:The experiment was performed in DME Center of Clinical Pharmacological Institute Affiliated to Guangzhou University of Traditional Chinese Medicine from January to August 2003, in which, 40 healthy male SD rats of 3 months old were employed and NG108-15 cell line was frozen-preserved.into the control, original formula group (No. 1 group) [shechuanzi (Cnidium monnieri (L.) Cuss), gouqizi (Lycium barbarum L.), renshen (Panax ginseng C.A.Mey.), heshouwu (Polygonum multiflorum Thunb.), danpi (Paeonia Suffruticisa Andr.) and bingpian (Borneolum)], kidney replenishment group (No.2 group) [shechuanzi (Cnidium monnieri (L.) Cuss), gouqizi (Lycium barbarum L.), etc.], group for benefiting qi and nourishing blood (No.3 group)[renshen (Panaz ginseng C.A.Mey), zhishouwu (Polygonum multiflorum Thunb.), etc.] and group with bingpian (Borneolum) removed (No.4 group)[Panax ginseng C.A.Mey.], heshouwu (Polygonum multiflorum Thunb.) and danpi (Paeonia Suffruticisa Andr.)], 8 rats in each group. The concentrated Chinese herbal solutions of every group were applied at 10 μL/g (equal to 6 g/kg of raw herbs) for gastric infusion successively,continuously for 1 month. In the control, the physical saline solution of equal dosage was used for infusion. Two hours after the last gastric infusion in rats of each group,the blood was collected from heart after anesthesia and the serum was sepaNG108-15 cell cultured in vitro was divided into 6 groups. In the control and model group, normal rat serum was contained in proliferated culture solution. In the rest 4 groups, the drug serum of No. 1 group and 3 sub-groups was contained.Simultaneously, beta-amyloid protein 25-35 in each hole was prepared to the terminal concentration 5 μmol/L (except in the control) and the culture went on for 48 hours.MAIN OUTCOME MEASURES:MTT method was used to determine proliferated number and survival rate of cells. Simultaneously, the ratio of neurite cells to total cell count and average length of neurit were determined.icantly than the control (0.520±0.022, 0.665±0.037, P < 0.01), and that in every drug serum group was higher than model group, of which, the result vival rate of differentiated cells: That in model group was lower significantly than the control (58.4%, 100%) and that in every drug serum group was higher than model group, of which, the result in No.4 group was the most tal cell count: That in model group was lower significantly than the control [(42.95±11.42)%, (58.75±12.84)%, P < 0.01] and that in every drug serum group was higher than model group, of which, the result in No.4 group was rite: That in model group was shorter significantly than the control [(356.0 ±109.0), (493.8±133.0) μm, P < 0.01] and that in every drug serum group was longer than model group, of which, the result in No.4 group was the most significant [(486.8±79.2) μm, P < 0.01].CONCLUSION: The drug serum in all of bushen yizhi formula and every subgroup inhibits at certain extent the injury of beta-amyloid protein 25-35 to NG108-15 cell, but the results of each group are various. The protection of drug serum to the cell in every group is in the sequence from strong to weak as group with bingpian removed > original formula group > kidney replenishment group > group for benefiting qi and nourishing blood. It is to expect a further study on the efficacy of group with bingpian removed.
6.Study on bacteria-inhibiting effect of amino acids
Yuekai CHEN ; Jun XU ; Yunbo QU ; Guoping XI ; Min ZHANG
Chinese Journal of Biochemical Pharmaceutics 2001;22(1):31-32
Purpose The aim is to study the inhibiting effect s of nineteen amino acids to E.coli,St.aureus and B.Subtilis.Metho ds The qualitative inhibition of nineteen amino acids on E.coli ,St.aureus and B.Subtilis was determined using filter papers treated by saturated amino acids placed on the bacteria medium dish.The quantitative inhibi ting effects of the amino acids which were proved to have inhibition to these th ree bacteria were determined in the bacteria LB medium with different concentrations of amin o acids.Results It was indicated that cysteine was used to in hibit St.aureus.The best inhibiting time was 6 hours,and the optimum of concentrat ion was 0.625% in which the inhibiting rate was 92.62%.Conclusion Cysteine could inhibit St. aureus.
7.Effects of Xuefu Zhuyu Decoction on the Morphological Features of Vascular Endothelial Cells Injured by Serum of Blood-stasis-syndrome Rabbits
Qi WANG ; Yunbo CHEN ; Shilong LAI ; Weixiong LIANG ; Zehuai WEN
Journal of Guangzhou University of Traditional Chinese Medicine 1999;0(02):-
Objective To observe the protective effect of Xuefu Zhuyu Decoction (XZD) on vascular endothelial cells (VEC) injured by the serum of blood_stasis_syndrome ( BSS) rabbits. Methods The morphological features of VEC cultured with norma l rabbit serum (Group A), BSS rabbit serum(Group B), the serum of normal rabbit medicated with XZD(Group C) and the serum of BSS rabbit medicated with XZD(Group D) were observed respectively under light microscope and electron microscope. Results Under light microscope, VEC was shrinked, intercellular space widene d and cytoplasm contained dark granules in Group B. The change of intercellular space was not obvious in Group D. Under electron microscope, pinocytotic vesicle s increased, rough endoplasmic reticulum (RER) decreased and dilated, mitochondr ia became smaller and indistinct and only a small amount of microvilli appeared on cell membrane with top expanded and merged in VEC of Group B as compared with Group A and Group C. Less RER but no dilatation and fewer microvilli without to p merged were found in VEC of Group B. Conclusion The serum of BSS rabbits c an injury normal VEC cultured in vitro and XZD exerts a certain protective effe ct on VEC injured by the serum of BSS rabbits.
8.Validation of the silencing site of rhesus monkey P21 gene in COS-7 cells
Yuhan LI ; Jingfen SU ; Chen ZHANG ; Liang SHI ; Yunbo LIU
Acta Laboratorium Animalis Scientia Sinica 2015;(3):297-300
Objective To screen the effective silencing targets of P21 gene at the cellular level in rhesus monkey . Methods To detect the expression of P21 gene in COS-7 cells ( derived from the kidney of African green monkey , Cerco-pithecus aethiops).Four small hairpin RNA (shRNA) sequences targeting rhesus monkey P21 gene were designed and in-serted into lentivirus-based gene silencing constructs FUGW-TDT.The vectors were transfected into COS-7 cells respective-ly.The suppression of P21 mRNA was detected by real-time PCR, and the expression of P21 protein was detected by West-ern blot assay .Results Four gene-silencing sequences were screened that lied in 541-561 bp, 542-562 bp, 215-239 bp, and 624-648 bp of the rhesus monkey P21 mRNA.Their silencing rate was (91.82 ±3.21)%, (82.47 ±2.48)%, (81.31 ±2.69 )% and ( 87.35 ±4.59 )%, and the protein expression was ( 11.97 ±0.70 )%, ( 20.22 ±0.65 )%, ( 23.21 ± 0.63)%and (14.42 ±0.86)%, respectively.Conclusions Four effective silencing target sequences are screened at cel-lular level , which can be used in gene silencing research of rhesus monkeys .
9.The early dynamics study of cerebrospinal fluid bacterial in a rabbit model of bacterial meningitis
Kaijin XU ; Huaying WANG ; Hongzhen JU ; Yunbo CHEN ; Zeqing WEI ; Yongtao LI ; Jifang SHENG ; Lanjuan LI
Chinese Journal of Infectious Diseases 2010;28(5):267-272
Objective To study the dynamics changes of cerebrospinal fluid (CSF) bacterial load within 48 h after infection in a rabbit meningitis model, and provide information for diagnosis,treatment and prognosis of this disease. Methods Taking New Zealand white rabbit as the study object, meningitis model was established via cerebellar cistern puncture with different concentrations of Escherichia coli (E. coli) and Staphylococcus aureus (S. aureus) to explore the relationship between the mortality of animals and the subarachnoid inoculation dosage. The dynamics study of CSF bacterial load was conducted with proper inoculation bacterial dosage. Forty-eight rabbits were separated into four groups (12 each group): E. coli meningitis model group, E. coli meningitis + ceftriaxone treated group, S. aureus meningitis model group and S. aureus meningitis + vancomycin treated group. At 0,12, 24, 36 and 48 h of inoculation, CSF and blood samples were obtained for CSF bacterial quantitative culture, CSF leukocyte count and peripheral blood leukocyte count. Finally, the relationships between the early mortality of animals, the efficacy of antibiotics, CSF leukocyte counts and the dynamics changes of CSF bacterial load were analyzed in the bacterial meningitis rabbit model.The CSF bacterial load and the white blood cell count curve were compared by analysis of covariance (ANOVA). Correlation test was done using correlate partial analysis. Results The relationship between subarachnoid inoculation dosage and the mortality of rabbits presented S-curve correlation.The bacterial load in subarachnoid space peaked in 12-24 h after infection and then gradually decreased. Effective antibiotic therapy could significantly speed up the decline of this process. There were significantly different between E. coli meningitis model group and E. coli meningitis+ceftriaxone treated group (F= 27. 10, P<0. 01), between S. aureus meningitis model group and S. aureus meningitis + vancomycin treated group (F=5. 97, P = 0. 016). There was a positive correlation between CSF bacterial load and CSF leukocyte count in E. coli and S. aureus meningitis model groups (r=0. 89, 0.84, respectively; P = 0.046, 0.049, respectively). Conclusions In the treatment of bacterial meningitis, effective and sufficient antibiotics should be used as soon as possible to control the CSF bacterial load and reduce the mortality. The CSF leukocyte count can be used as indicator of CSF bacterial load and guide the antibiotic treatment in clinical bacterial meningitis.
10.Bushen Huoxue Lishi Category TCM Compound in the Treatment of Chronic Prostatitis:A Systematic Re-view
Hongzhi GUO ; Yunbo LIU ; Mingyue ZENG ; Peining NIU ; Gangliang JIAO ; Qiang CHEN ; Yong ZHU ; Qingqi ZENG
China Pharmacy 2016;27(30):4241-4244
OBJECTIVE:To systematically review the efficacy and safety of the Bushen huoxue lishi category TCM compound in the treatment of chronic prostatitis,and provide evidence-based reference for clinical treatment. METHODS:Retrieved from VIP Database,Wanfang Database,CJFD and CBM,randomized controlled trials(RCT)about Bushen huoxue lishi category TCM com-pound preparation (test group) versus conventional Western medicine (control group) in the treatment of chronic prostatitis were collected. Meta-analysis was performed by Rev Man 5.3 software after data extraction and quality evaluation. RESULTS:Totally 22 RCTs were enrolled,involving 1 863 patients. Results of Meta-analysis showed the total effective rate [OR=4.46,95%CI(3.40, 5.84),P<0.001],total scores of chronic prostatitis symptoms[MD=-3.62,95%CI(-5.21,-2.04),P<0.001] and lecithin count [MD=7.58,95%CI(2.15,13.01),P=0.006] in test group were significantly higher than control group,prostatic fluid white blood cell count [MD=-1.68,95%CI(-3.26,-0.10),P=0.04] was significantly lower than control group,with significant differenc-es. CONCLUSIONS:Bushen huoxue lishi category TCM compound has good efficacy in the treatment of chronic prostatitis.