1.The effect of AD injection on immune functions in tumor-bearing mice
Chinese Journal of Immunology 2010;26(3):224-227
Objective:In order to study the antitumor effect of AD injection and its immunological mechanisms.Methods:Liver cancer H22-bearing mice were treated with AD injection and 7 days later to observe the effect of AD injection on the inhibitory rate of tumor,indexes of thymus and spleen,lymphocytes transformation function,phagocytosis of the peritoneal macrophage,activity of TNF,the level of IL-2 and IFN-α in serum.Results:AD injection could inhibit the growth of tumor in bearing mice significantly and promote the transformation function of lymphocytes and enhance the peritoneal macrophage phagocytosis and the activities of TNF.AD injection could improve the level of IL-2 of spleen cells and IFN-α in serum.Conclusion:These results suggested that AD injection could obviously inhibit the growth of tumor and enhance the immune functions in bearing mice.
2.Role of solasodine hydrochloride in AS2O3 induced HeLa cells apoptosis as well as its effect on cell telomerase activity in vitro
Jin-Xia, AI ; Liang, LIU ; Ping, WANG
Chinese Journal of Endemiology 2011;30(3):279-283
Objective To study whether solasodine hydrochloride (SBHL) could enhance the effect of arsenic trioxide in inducing apoptosis and affecting telomerase activity in cervical cancer HeLa cells. Methods Using cell culture methods, cervical cancer HeLa cells were cultured in vitro. The optimal concentration of SBHL was determined by MTT method from 0, 10, 20, 40, 80, 160, to 320 μmol/L. HeLa cells were grown in improved RPMI1640 supplemented respectively with arsenic trioxide(5 μmol/L As2O3), As2O3(5 μmol/L)+ SBHL( 40 μmol/L) and none (control group). The growth morphology of HeLa cells was observed under phase contrast microscopy after culture for 24, 48, and 72 h. Apoptosis of HeLa cells was determined under transmission electronic microscopy. The method of MTT was used to study the cell survival percentage. The technique of flow cytometry was used to measure cell cycle and cell apoptosis percentage. The method of tartrate-resistant acid phosphatase-enzyme linked immunosorbent assay (TRAP-ELISA) was used to determine telomerase activity of HeLa cells. Results Under phase contrast microscopy, in control group HeLa cells were round, densely packed; in As2O3 group the numbers of the cells were less, cell spacing increased; in As2O3 + SBHL group the cells shrinked significantly, nuclear fragmented as a petal-like, gap became larger. Under transmission electronic microscopy, there were rich microvillus on the cell surface in control group, cell intervals clear, immature connections, and the intervals did not close. The structure of the mitochondria in the cytoplasm was integrated. Most of the chromatin in the nucleus were, euchromatin and characteristics of apoptosis with heterochromatin increased and the chromatin condensed into masses, on the boundary of nuclear membrane. The microvillud on the cell surface were ruptured and decreased in As2O3 + SBHL group. The chromatin condensed into masses. The formation of apoptotic bodies was observed. The difference was statistically significant between groups in cell survival percentage at 24, 48, 72h(x2 = 10.39 , 13.88 , 17.21,respectively, all P < 0.05). Cell survival percentage in SBHL + As2O3 group (52.80%) was significantly less than that of As2O3 group(77.51%, x2 = 9.29, P < 0.05) at 72 h. In cell cycles, the difference was statistically significant between groups in C1 phase and S phase(F = 7.46,22.14, all P < 0.05), respectively. Compared with , control group[ (41.57 ± 1.56)%, (50.45 ± 2.37)%], cell percentages in S phase in As2O3 + SBHL group[(20.06 ± 4.98)%] and As2O3 group[(27.10 ± 5.32)%] were decreased(P< 0.05 or < 0.01), while cell percentage in C1 phase was increased[(58.70 ± 5.18)%, (69.67 ± 4.17)%, P< 0.05 or < 0.01]. The difference was statistically significant between groups in apoptotic percentage of HeLa cells (F = 4.01, P < 0.05). Compared with control group[ (1.18 ± 1.40)%], apoptosis percentage was significantly increased in As2O3 + SBHL group and As2O3 group [(21.08± 1.22)%, (6.04±2.53)%, P< 0.05 or < 0.01], respectively, and As2O3 + SBHL group was higher than As2O3 group(P < 0.01). The difference was statistically significant between groups in telomerase activity (F = 21.28, P< 0.05). Telomerase activity was inhibited in As2O3 group(1.214 ± 0.621) and As2O3A + SBHL group(0.865 ± 0.284) compared to control group (2.107 ± 0.057, all P < 0.05), and telomerase activity in As2O3 + SBHL group was lower than that of As2O3 group (P < 0.05). Conclusions SBHL enhances the effect of As2O3 in inducing apoptosis in HeLa cells, which is related to its inhibiting telomerase activity in HeLa cells.
3.Effect of aplysin on the proliferation and apoptosis in human gastric cancer cell SGC-7901
Ying LIU ; Hui LIANG ; Ai SU ; Juan HE ; Hong YU
Chinese Pharmacological Bulletin 2010;26(3):333-337
Aim To investigate the effects of Aplysin on the inhibition of gastric cancer cell in vitro .Methods MTT assay was used to examine the inhibition of gastric cancer cell 1ine SGC-7901 by Aplysin in different concentrations and at different times.The morphologic changes and the apoptosis of SGC-7901 was observed by inverted microscope and Hematoxylin-Eosin(HE)staining.Reverse transcriptase polymerase chain reaction(RT-PCR)assay was used to detect the changes of COX-2 mRNA expressions.Results Aplysin could decrease the proliferation significantly in a dose-dependent manner in SGC-7901 cells.When treating SGC-7901 with Aplysin in concentration of 120, 240 mg·L~(-1) for 24 h, the growth of the cell was obviously inhibited observing by inverted microscope.Aiso, when treating with the same concentration for 18 h, its chromatin became crimpled and breakdown, as well as cell shrinkage and apoptotic bodies formation when using HE staining.The apoptotic rates(%)of SGC-7901 was(15.0±2.12)%, (18.4±2.3)%, respectively, which was significantly higher than(1.4±0.55)% that in control group(P <0.01).60、120、240 mg·L~(-1) Aplysin could not effectively inhibited the mRNA expressions of COX-2(P >0.05).Conclusions Aplysin can inhibit the proliferation and induces apoptosis of SGC-7901 cells.
4.Effects of Aplysin on ethanol-induced oxidative damage in rat primary hepatocytes
Ai SU ; Hongyan ZHU ; Hongwei XU ; Ying LIU ; Hui LIANG
Chinese Pharmacological Bulletin 2016;(2):251-257
Aim To investigate the protective effects of Aplysin on ethanol-induced oxidative damage in rat pri-mary hepatocytes. Methods Rat primary hepatocytes were obtained via the portal vein collagenaseⅣin situ perfusion technique followed by a Percoll density gradi-ent centrifuge. MTT test was used to determine the op-timum dose of Aplysin and ethanol, and detect the cell vitality in primary hepatocytes. Supernatants of primary hepatocytes were harvested to measure AST and LDH level, and the SOD, GSH-PX activities and MDA con-tent in primary hepatocytes were observed. Flow cy-tometry was used to detect the cell apoptosis rate. DNA damage in primary hepatocytes was detected by single-cell gel electrophoresis assay. The level of mitochon-drial membrane potential in primary hepatocytes was tested by fluorogenic probe JC-1 . The CYP2 E1 activity in primary hepatocytes was detected by colorimetry. The proteins of CYP2 E1 were detected by Western blot. Results 300 mmol·L-1 dose of ethanol and 30 mg·L-1 dose of Aplysin were the optimal dosages and were used in the subsequent experiments. Hepatocyte vitality was significantly increased in Aplysin group compared to that in ethanol group, and Aplysin inhibi-ted the release of AST and LDH(P<0. 05). For Apl-ysin treatment group, the activities of hepatocyte SOD and GSH were significantly increased, and MDA was markedly lowered as compared with those in ethanol group( P <0. 05 ) . Aplysin could alleviate hepatocyte apoptosis significantly, and hepatocyte DNA damage rates of Ⅱ ~Ⅲ level and Ⅳ level were significantly lowered in Aplysin treatment group as compared with those in ethanol group, and Aplysin had evident im-provement in alcohol induced mitochondria damage of hepatocyte. Primary hepatocyte activities and protein expression of CYP2 E1 were markedly lowered in Aply-sin treatment group as compared with those in ethanol group(P<0. 05). Conclusion Aplysin has protective effects on liver oxidative damage induced by alcohol of primary cultured rat hepatocytes by blocking CYP2 E1 activation, relieving oxidative stress, and sharpening the oxidation resistance ability.
5.Biological Characteristics of Human Colon Cancer Cell Line CW-2 Stem Cells
Mei LIU ; Liang AI ; Mi CHEN ; Hao LONG ; Shaolin LI
Journal of China Medical University 2010;(6):417-419
Objective To isolate cancer stem cells from human colon cancer cell line CW-2,and observe the biological characteristics.Methods The percentage of CD44+EpCAM+ cancer stem cells which were isolated by multi-combination method was evaluated by flow-cytometry.The biological characteristics of the stem cells were analyzed by cell cycle analysis,in vitro invasion assay and in vivo tumorigenicity assay.Results The percentage of CD44+EpCAM+ cancer stem cells was 89.57%,and their capabilities of generation,invasion,tumorigenicity were higher than non-cancer stem cells(P 0.05).Conclusion CD44+EpCAM+ cancer stem cells showed low generation,high invasiveness,high tumorigenesis,and they could be isolated by multi-combination.
6.Screening and Identification of Broadspectrum Bacteriocin-producing Lactic Acid Bacterium
Ai-Qing ZHANG ; Shu-Liang LIU ; Ling AO ;
Microbiology 1992;0(04):-
The text has isolated 267 strains LAB from the traditional fermentative food in Sichuan. Using agar plate proliferation experiment and double-layer agar plate proliferation experiment (eliminating the effects of organic acid and H_2O_2, decreased after treatment with trypsin and papain) to screen a LAB of Bacteriocins P158 which has antibacterial action to Escherichia coli, Staphylococcus aureus, Micrococcus luteus, Pseudomonas aeruginosa, Bacillus subtilis. P158 was isolated from fermented glutinous rice. Through detection of its appearance, physiological and biochemical characteristics and 16S rDNA gene sequence homology analysis, it was identified as Lactobacillus plantarum.
7.Preliminary study of odor change mechanism in Crataegi fructus stir-fried process based on correlation analysis.
Liang LI ; Shi-Long YANG ; Yu-Jie LIU ; Yun-Wei WSNG ; Lian ZHONG ; Li AI
China Journal of Chinese Materia Medica 2014;39(17):3283-3286
In order to investigate the mechanism, the correlation between the odor change in Crataegi Fructus stir-fried process and 5-HMF were studied. Required samples were retrieved from Crataegi Fructus stir-fried process. Statistical quality control (SQC) was used to analyze the response values acquired by the electronic nose. At the same time, the content of 5-HMF was detected by high performance liquid chromatography (HPLC). Correlation analysis was used to analyze the relationship between the above two. Experimental results showed that SQC model established by response values of all samples could show the change law of odor in Crataegi Fructus stir-fried process and changes of 5-HMF content was dropped after the first increase. Correlation analysis showed that the odor change in Crataegi Fructus stir-fried process and 5-HMF were significantly correlated (P < 0.05). Sugar degradation reaction and the Maillard reaction may be one of the mechanisms of the odor change in Crataegi Fructus stir-fried process.
Chromatography, High Pressure Liquid
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Crataegus
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chemistry
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Furaldehyde
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analogs & derivatives
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analysis
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Hot Temperature
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Odorants
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analysis
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Plant Extracts
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analysis
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Technology, Pharmaceutical
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methods
8.The Establishment of NMR Characteristic Fingerprint of Cordyceps Sinensis and Its Identification Study
Gang CHEN ; Liang HUANG ; Wenjia LI ; Zeping ZHAN ; Libo XIN ; Zhong AI ; Zhengming QIAN ; Guozhu LIU
World Science and Technology-Modernization of Traditional Chinese Medicine 2014;(11):2371-2379
The inundation of Cordyceps sinensis counterfeits in the market makes it difficult to identify. In this study, 21 batches of wild C. sinensis from 3 different regions, 30 batches of naturally cultured C. sinensis and 4 kinds of counterfeits extracted by methanol and water were analyzed using NMR technology. 9 characteristic peaks were defined as quantitative criterion after comparison, and NMR fingerprints of C. sinensis were established. According to the result it is highly similar between naturally cultured C. sinensis and wild ones by comparing their NMR fingerprints. However, NMR spectra of four kinds of adulterants showed differences with C. sinensis. The result also showed that NMR fingerprint of C. sinensis are highly characteristic and specific. The NMR characteristic fingerprint of wild C. sinensis was consistent with the naturally cultured C. sinensis, and it indicated that the chemical constituents of wild C. sinensis and naturally cultured C. sinensis are nearly the same.
9.Treatment of the local masses complicated after the periorbital fat transplantation
Weizhong LIANG ; Hongmei AI ; Yingyong WANG ; Chang LIU ; Kuoyuan WANG ; Zuojun ZHAO
Chinese Journal of Medical Aesthetics and Cosmetology 2016;22(2):84-86
Objective To evaluate the causes of local masses found after periocular fat grafting,and the way of treatment in order to avoid the medical dispute.Methods In 37 patients with local mass being repaired,15 patients with superficial bulges were treated through surgical removal and as piration;10 patients with focal masses were treated with aspiration;12 patients with bulges close to eyelid margin and failure after the first two treatment methods were treated through incision.Results Histopathologic findings showed the characteristic features of lipogranuloma.Chronic inflammatory cells were infiltrated.Numerous,variable-sized lipid vacuoles were surrounded by histiocytes and for eign body-type giant cells.There were areas of fibrosis and fat necrosis.35 cases were followed-up for 3 months to 1 years.Both doctors and patients were satisfied with results in 27 cases,basically satisfied in 6 cases,but not satisfied in 2 cases.Two patients were lost to follow up.Conclusions The special complication of masses in the periocular fat grafting operation can be greatly improved with the personalized therapy.
10.Pilot study on hypotensive effect of combination of 0.004% travoprost with 2% pilocarpine on normal albino rabbit
Shun-hua, ZHANG ; Jia-liang, ZHAO ; Xiao-li, LIU ; Ai-ling, BIAN
Chinese Journal of Experimental Ophthalmology 2012;30(9):774-778
Background It has been clarified that topical administration of cholinomimetic drug or prostaglandin analogus can effectively low intraoeular pressure (IOP) in primary angle-closure glaueoma and angleopen glaucoma,respectively.The two eyedrops are often combined clinically to treat glaueomatous eye.But their effect mechanism on ciliary muscle is different.It is neeessarv to prove the clinical efficacy of combination of these two drugs.Objective This study attempted to compare hypotensive effect of individual and combinative use of 2%piloearpine and 0.004% travoprost in normal rabbit eyes.Methods Thirty normal albino rabbits were randomly divided into three parallel groups.Lateral eye of each rabbit was appointed as the experimental eye and the fellow eye was used as control eye.2% Pilocarpine eyedrop was instilled three times daily and 0.004% travoprost eyedrops was used once per night in the pilocarpine and travoprost group.These two drugs were combined in the combination group.The normal saline solution was used in the control eyes IOP was measured bilaterally with Perkins application tonometer at 8:00 am on the day before and 1,2,4,8,14,24 days after the administration of eyedrops.The use of the animals complied with the Regulations for the Administration of Affairs Concerning Experimental Animals by State Science and Technology Commission.Results The IOP was significantly lower on 1 day after use of eyedrops than baseline IOP in the experimental eyes in the pilocarpine group,travoprost group and combination group,but no obvious change in IOP before and after use of eyedrops in the control eyes.There was no significant difference in the baseline IOP in both experimental eyes and control eyes,however,signifieant differences were seen in various time points after administration of eyedrops between experimental eyes and control eyes(P<0.05).Compared with baseline IOP,IOP declined by 17.5%-22.0% in pilocarpine group,23.8%-26.4% travoprost group,27.6%-32.0% in the combination group.For normal eye,piloearpine+travoprost showed a more power lowing IOP effect.Conclusions Hypotensive effect of combinative use of 2% pilcoarpine and 0.004% travoprost is strongest in comparison with individual use of pilocarpine or travoprost,but less than the statistical sum of individual use of these two drugs in a 24-day duration in normal eye of albino rabbit.