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.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.
3.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.
4.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.
5.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.
6.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 .
7.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.
8.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
9.The influence of anesthesia and means of postoperative pain control on T lymphocyte subtypes of blood in patients with rectal cancer
Yalan LI ; Yunbo CHEN ; Shihuang MO ; Xuemei PENG ; Xiaoping WANG ; Changren ZHOU
Chinese Journal of Pathophysiology 1989;0(06):-
AIM: To evaluate the influence of anesthesia and different means of postoperative pain control on the T-lymphocyte during the perioperative period in patients with rectal cancer.METHODS: 40 adult patients, aged 65 or older, of American Society of Anesthesiologists (ASA) class 2-3 were divided into two groups according to the type and means of postoperative pain managements. Group Ⅰ (n=20) received intravenous anesthesia and patient controlled analgesia(PCA), fentanyl (13 ?g/kg) for post pain; group Ⅱ (n=20) received intravenous anesthesia plus lumber epidural anesthesia and epidural PCA of morphine 5 mg plus ropivacaine 100 mg for post operative pain. Blood samples from internal jugular vein were obtained before surgery, at the completion of surgery and 24, 48, and 120 h post surgery for detecting CD3+, CD4+, CD4/CD8 counts of peripheral T-lymphocytes. In addition, blood cortisol level and pain intensity were assessed by visual analogue score (VAS)at each time point. RESULTS: Baseline(before anesthesia) values of CD3+,CD4+, CD4/CD8 in patients were messured and there was a significant decrease of all these values from completion of surgery to 48 h after surgery in both groups (P
10.Effects of Blood-stasis-syndrome Rabbit Serum on Gene Expression of Activating Factors in Cultured Vascular Endothelial Cells
Yunbo CHEN ; Mengjun HOU ; Qi WANG ; Weixiong LIANG ; Zehuai WEN ; Shilon LAI
Journal of Guangzhou University of Traditional Chinese Medicine 1999;0(02):-
Objective To explore the molecular mechanism of blood_stasis syndrome (BSS) by studying the changes of gene expression of endothelin (ET) and constit utive nitric oxide synthase (cNOS) in vascular endothelial cells (VEC) cultured w ith BSS rabbit serum. Methods ET_1 mRNA expression and cNOS mRNA expression were analyzed by semi_quantitative reverse transcription and polymerase chain re action method. Results ET_1 mRNA expression level was increased and cNOS mRN A expression level was decreased in VEC cultured with BSS rabbit serum (Group A) as compared with VEC cultured with normal rabbit serum (Group B) or without rab bit serum (Group C) (P