Effect of Toddalia asiatica Alcohol Extract on Apoptosis of Non-small Cell Lung Cancer A549 Cells Based on Autophagy Pathway / 中国实验方剂学杂志

ObjectiveTo study the effects of Toddalia asiatica alcohol extract on autophagy and apoptosis of non-small cell lung cancer A549 cells， and to explore its possible mechanism. MethodA549 cells were cultured in vitro. Cell counting kit-8 （CCK-8） was used to detect the proliferation of A549 cells， and cell survival rate was calculated to screen the drug concentration. The apoptosis in each dose group and that after the use of 3-methyladenine （3-MA）， an autophagy inhibitor， were detected by flow cytometry combined with Annexin V-FITC/PI double staining. Western blot was used to detect the expression levels of apoptosis-related proteins such as B cell lymphocytoma-2（Bcl-2）， Bcl-2-associated X protein（Bax）， microtubule-associated protein 1 light chain 3 （LC3）， cleaved cysteinyl aspartate-specific protease-3 （cleaved Caspase-3）， activated poly （Adenosine diphosphate） ribonucleotide polymerase （cleaved PARP1）， PARP1， activated death activator （t-Bid）， Bid， and ubiquitin-binding protein p62 in each group and those after the use of 3-MA. ResultCompared with the conditions in the control group， the cell survival rate in 0.25 g·L-1 group （P<0.05）， and 0.5， 1， 2， 4 g·L-1 groups （P<0.01） was decreased after 24 h intervention. Additionally， the cell survival rate was reduced in a concentration-dependent manner at 48 h and it was less than 10% at 4 g·L-1 （P<0.01）. Compared with the conditions in the control group， the total apoptosis rate in 0.5 g·L-1 group was increased （P<0.05）， and the apoptosis rate in 1 and 2 g·L-1 groups was also increased （P<0.01）. Compared with the 2 g·L-1 group and 3-MA group， the 3-MA combined with T. asiatica alcohol extract had significantly decreased apoptosis rate （P<0.01）. Compared with the conditions in the control group， elevated expression of pro-apoptotic proteins cleaved PARP1， Bax and t-Bid in 1 and 2 g·L-1 groups （P<0.05， P<0.01）， and reduced expression of Bid in the 2 g·L-1 group （P<0.01） were found. Compared with the conditions in the control group， the expression of anti-apoptotic protein Bcl-2 （P<0.05， P<0.01） and the level of p62 （P<0.01） were down-regulated in 0.5， 1， 2 g·L-1 groups， while the level of LC3 Ⅱ protein was up-regulated （P<0.01）， with certain concentration dependence. ConclusionT. asiatica alcohol extract could significantly inhibit the proliferation of A549 cells， which might be related to promoting autophagy and inducing apoptosis.

Progress in the molecular mechanism of KaiA regulating cyanobacterial circadian clock / 生物工程学报

The cyanobacterial circadian clock has three relatively independent parts: the input path, the core oscillator, and the output path. The core oscillator is composed of three clock proteins: KaiA, KaiB, and KaiC. The interactions among these three proteins generate a rhythmic signal and convey the input signals to the output signals to maintain the accuracy and stability of the oscillation of downstream signals. Based on the cyanobacterial circadian clock and the structure, function, and interaction of the clock proteins of the core oscillator, combining the recent results from our laboratory, this review summarized the recent progresses of the molecular mechanism of KaiA in regulating KaiC's enzymatic activity, mediating phase reset of the oscillator, and competing with CikA for the binding site of KaiB.

Molecular mechanism underlying the inhibitory effect of spermine oxidase inhibitor SI-4650 on proliferation of a human malignant melanoma cell line A375 / 中华皮肤科杂志

Objective@#To evaluate the effect of spermine oxidase (SMO) inhibitor SI-4650 on the proliferation of a human malignant melanoma cell line A375, and to explore its molecular mechanism.@*Methods@#Some cultured A375 cells were divided into 6 groups to be treated with SI-4650 at concentrations of 0, 10, 20, 40, 80 and 160 μmol/L respectively for 24, 48 and 72 hours, and methyl thiazolyl tetrazolium (MTT) assay was performed to evaluate changes in cellular proliferative activity. According to the cellular proliferative activity, 3 concentrations (0, 40, 80 μmol/L) were screened out. Some A375 cells were divided into 3 groups to be treated with 0 (control group) , 40 and 80 μmol/L SI-4650 for 48 hours. Chemiluminescence assay was conducted to detect the SMO activity in A375 cells, high-performance liquid chromatography (HPLC) analysis to determine the polyamine content in A375 cells, flow cytometry to analyze the cell cycle and detect the apoptosis, and Western blot analysis to determine the protein expression of apoptotic marker proteins Bax and c-PARP, inhibitor of apoptosis protein Bcl-2, and autophagy marker proteins Beclin-1 and LC3-Ⅱ. Statistical analysis was carried out by using one-way analysis of variance for comparison of means among several groups, and by using Student-Newman-Keuls (SNK) -q test for multiple comparisons.@*Results@#MTT assay showed that there was a significant difference in the proliferative activity of A375 cells after the treatment with different concentrations of SI-4650 for different durations (F = 977.23, 5.16 respectively, both P < 0.001) . Significant differences were observed in the SMO activity in A375 cells (F = 242.58, P < 0.001) , spermine and the total polyamine content (F = 338.02, 2 931.07 respectively, both P < 0.001) , proportion of S-phase cells (F = 31.66, P < 0.001) , proportion of apoptotic cells (F = 100.68, P < 0.001) , expression of apoptosis-related proteins Bax, c-PARP and Bcl-2 (F = 35.51, 730.11, 27.54 respectively, all P < 0.001) , and expression of autophagy marker proteins Beclin-1 and LC3-Ⅱ (F = 35.87, 425.04 respectively, P < 0.001) among the control group, 40- and 80-μmol/L SI-4650 groups. Compared with the control group, the 40- and 80-μmol/L SI-4650 groups showed significantly lower SMO activity (luminous intensity: 61 432.85 ± 2 620.92, 43 337.35 ± 1 221.25 respectively, both P < 0.05) , lower spermine (1.97 ± 0.007, 1.88 ± 0.006 respectively, both P < 0.05) and total polyamine content (3.18 ± 0.03, 2.81 ± 0.01 respectively, both P < 0.05) , higher proportions of S-phase cells (27.61% ± 2.05%, 31.58% ± 1.45% respectively, both P < 0.05) and apoptotic cells (27.61% ± 2.05%, 31.58% ± 1.45% respectively, both P < 0.05) , higher expression of apoptotic marker proteins Bax (0.83 ± 0.12, 1.18 ± 0.16 respectively, both P < 0.05) and c-PARP (0.32 ± 0.002, 0.79 ± 0.035 respectively, both P < 0.05) and autophagy marker proteins Beclin-1 (1.00 ± 0.007, 1.14 ± 0.003 respectively, both P < 0.05) and LC3-Ⅱ (0.31 ± 0.001, 0.98 ± 0.003 respectively, both P < 0.05) , and lower expression of inhibitor of apoptosis protein Bcl-2 (0.65 ± 0.09, 0.12 ± 0.002 respectively, both P < 0.05) .@*Conclusion@#SI-4650 can inhibit the proliferation of A375 cells, likely by interfering with polyamine metabolism and inducing cell cycle arrest, apoptosis and autophagy.

Molecular mechanism underlying the inhibitory effect of spermine oxidase inhibitor SI-4650 on proliferation of a human malignant melanoma cell line A375 / 中华皮肤科杂志

Objective To evaluate the effect of spermine oxidase(SMO)inhibitor SI-4650 on the proliferation of a human malignant melanoma cell line A375, and to explore its molecular mechanism. Methods Some cultured A375 cells were divided into 6 groups to be treated with SI- 4650 at concentrations of 0, 10, 20, 40, 80 and 160 μmol/L respectively for 24, 48 and 72 hours, and methyl thiazolyl tetrazolium(MTT)assay was performed to evaluate changes in cellular proliferative activity. According to the cellular proliferative activity, 3 concentrations (0, 40, 80 μmol/L) were screened out. Some A375 cells were divided into 3 groups to be treated with 0(control group), 40 and 80μmol/L SI-4650 for 48 hours. Chemiluminescence assay was conducted to detect the SMO activity in A375 cells, high-performance liquid chromatography(HPLC)analysis to determine the polyamine content in A375 cells,flow cytometry to analyze the cell cycle and detect the apoptosis, and Western blot analysis to determine the protein expression of apoptotic marker proteins Bax and c-PARP, inhibitor of apoptosis protein Bcl-2, and autophagy marker proteins Beclin-1 and LC3-Ⅱ. Statistical analysis was carried out by using one-way analysis of variance for comparison of means among several groups, and by using Student-Newman-Keuls (SNK)-q test for multiple comparisons. Results MTT assay showed that there was a significant difference in the proliferative activity of A375 cells after the treatment with different concentrations of SI-4650 for different durations(F=977.23, 5.16 respectively, both P<0.001). Significant differences were observed in the SMO activity in A375 cells(F=242.58, P<0.001), spermine and the total polyamine content(F=338.02, 2931.07 respectively, both P < 0.001), proportion of S-phase cells (F = 31.66, P < 0.001), proportion of apoptotic cells(F=100.68, P<0.001), expression of apoptosis-related proteins Bax, c-PARP and Bcl-2(F = 35.51, 730.11, 27.54 respectively, all P < 0.001), and expression of autophagy marker proteins Beclin-1 and LC3-Ⅱ(F = 35.87, 425.04 respectively, P < 0.001)among the control group, 40-and 80-μmol/L SI-4650 groups. Compared with the control group, the 40-and 80-μmol/L SI-4650 groups showed significantly lower SMO activity(luminous intensity:61432.85 ± 2620.92, 43337.35 ± 1221.25 respectively, both P<0.05), lower spermine(1.97 ± 0.007, 1.88 ± 0.006 respectively, both P<0.05)and total polyamine content (3.18 ± 0.03, 2.81 ± 0.01 respectively, both P < 0.05), higher proportions of S-phase cells (27.61％ ± 2.05％, 31.58％ ± 1.45％ respectively, both P < 0.05) and apoptotic cells (27.61％ ± 2.05％, 31.58％ ± 1.45％ respectively, both P < 0.05), higher expression of apoptotic marker proteins Bax(0.83 ± 0.12, 1.18 ± 0.16 respectively, both P < 0.05)and c-PARP(0.32 ± 0.002, 0.79 ± 0.035 respectively, both P < 0.05)and autophagy marker proteins Beclin-1(1.00 ± 0.007, 1.14 ± 0.003 respectively, both P < 0.05)and LC3-Ⅱ(0.31 ± 0.001, 0.98 ± 0.003 respectively, both P < 0.05), and lower expression of inhibitor of apoptosis protein Bcl-2(0.65 ± 0.09, 0.12 ± 0.002 respectively, both P<0.05). Conclusion SI-4650 can inhibit the proliferation of A375 cells, likely by interfering with polyamine metabolism and inducing cell cycle arrest, apoptosis and autophagy.

Effect of mesalazine combined Bifidobacterium triple viable in the treatment of ulcerative colitis / 中国生化药物杂志

Objective To explore the beauty of mesalazine combined with Bifid Triple viable bacilli clinical efficacy in the treatment of ulcerative colitis.Methods 72 patients with ulcerative colitis ( UC) in zhoushan maternal and child health hospital were selected and divided into experimental group and control group,36 cases in each group.The control group was treated with oral administration of melamine and the experimental group was treated with Bifidobacterium triple live bacteria on the basis of oral control group.The levels of superoxide dismutase (SOD), malondialdehyde (MDA) and reactive protein ( CRP) concentration in serum of patients were detected.Follow up observation and record of the experimental group and the control group to improve the symptoms, clinical efficacy and adverse reactions after treatment.Results After treatment,serum MDA, CRP concentrations of the two groups were significantly lower than those before treatment concentration, SOD concentration was significantly increased, the differences were statistically significant (P<0.05) , serum MDA, CRP levels in the experimental group were lower than the control group, the concentration of SOD was significantly higher than control group,the differences were statistically significant (P<0.05) .After treatment, abdominal pain, diarrhea, mucus and blood stool symptoms of two groups were significantly improved compared with before treatment, abdominal pain, diarrhea and bloody mucus symptoms in the experimental group were lower than the control group(P<0.05).Incidence of adverse reactions in the experimental group was significantly lower than the control group, the differences were statistically significant (P<0.05).Conclusion The efficacy of oral administration of methadiazole in combination with Bifidobacterium triple live bacteriain the treatment of ulcerative colitis is significant, Can reduce MDA, CRP concentration, increased SOD activity.

Inhibiting effect of DZNep on proliferation of human prostatic cancer PC3 cells by down-regulating EZH2 expression / 实用医学杂志

Objective To study the effects of S-adenosyl-homocysteine hydrolase inhibitor DZNep (3-Dea zaneplanocin A) on human prostatic cancer PC3 cells growth and further explore its potential value in anti-human prostatic cancer treatment.Methods MTT method was used to analyze the effects of EZH2 gene silencing,EZH2 over-expression and DZNep treatment on cell proliferation.Gene expression of EZH2,Bax and Bcl-2 in PC3 cells was detected with western blot.Results DZNep could significantly inhibit PC3 cell growth and induce apoptosis which was identified with Bax expression up-regulation and Bcl-2 expression down-regulation.EZH2 knock-down inhibited PC3 cells growth,and over-expression of EZH2 partially counteract the inhibitory effects of DZNep on PC3 cells growth.Conclusions DZNep can inhibit PC3 cells growth by targeting EZH2 inhibition which leads to endogenous apoptosis.DZNep has the potential value in the treatment of human prostatic cancer.

Immunogenic apoptosis of human Caski cervical cancer cells induced by Lidamy-cin / 中国免疫学杂志

Objective:To investigate effects of Lidamycin (LDM,C-1027) on the proliferation and immunogenic transform of human Caski cervical cancer cells and to provide the basic experiment data and theoretical supports for establishment of the new immu-notherapy method mediated by LDM. Methods:MTT was used for the analysis of cell proliferation;apoptosis rate was analyzed by flow cytometry;Western blot was used to analyze the effect of LDM on the expression of Bax and Bcl-2 in Caski cells;the Flow cytometry was used to detect the expression of Calreticulin ( CRT ) on the cell surface. Results: Lidamycin inhibited proliferation of Caski cells significantly in the time-and dose-dependent manners;The apoptotic cell ratio induced by 5 μg/L Lidamycin was 11. 5% ,Comparing with the control group, Lidamycin treatment increased Bax but decreased Bcl-2 contents significantly within Caski cells, it also significantly increased the expression of CRT on the cell surface of Caski cells from 2. 31% to 67. 2%. Conclusion: Lidamycin has pharmacological activity in inhibiting proliferation of the human cervical Caski cells and the underlying mechanism is related with inducing the intrinsic mitochondrial pathway of apoptosis. In the same time,Lidamycin can increase the expression of CRT on the cell surface,so it may have the ability to promote the immunogenic apoptosis of tumor cells.

OAZI-1 overexpression in mouse B16-F1 melanoma cells can effectively stimulate DC immune activity / 中国免疫学杂志

Objective:To analyze, at cellular level, whether the mouse B16-F1 melanoma cells with OAZI-1 overexpression could activate antigen-presenting cells and promote the phagocytotic and antigen-presenting efficiencies of mouse peritoneal macrophage and bone marrow derived DC on tumor cells. Methods:The plasmid pcDNA3. 1(+)/OAZI-1 was transfected into B16-F1 cells by Li-pofectamine2000 reagent. The positive clones with OAZI-1 overexpression ( B16/OAZI-1 ) were identified by Western blot assay and RT-PCR. Macrophages from abdominal cavity and DC from bone marrow were collected from BALB/c mouse. The B16-F1 cells transfected with the pcDNA3. 1(+) (B16/3. 1) were used as the control cells in this experiment. B16-F1 cells and macrophages were co-cultured for 4 h at a 1∶5 ratio and DC were co-cultured with B16-F1 cells at 1∶1 ratio for 4 h. And then the phagocytotic efficiencies were assayed by flow cytometry. DC were co-cultured with B16-F1 cells at 1∶1 ratio for 24 h and then the expression of mature DC surface marker molecules CD40,CD80,CD86 were determined by flow cytometry. The DC activated by the tumor cells were co-cultured with mouse spleen lymphocytes for 24 h, and then IFN-γ content in culture medium was analyzed by ELISA. Results: Phagocytotic assay showed that,compared to the control cells,the OAZI-1 overexpression in B16-F1 cells significantly enhanced the engulfment of B16-F1 cells by macrophages ( 24. 7% vs 53. 9% ) and DC ( 8. 2% vs 13. 8%) . When DC were co-incubated with OAZI-1 overexpressed B16-F1 for 24 h,the expression levels of CD40,CD80,CD86 on the DC surface,which were the molecular markers for matured DC,increased from 24. 2%,20. 8% and 16. 4% to 46. 8%,32. 5% and 36. 1% respectively. Co-culture of tumor-activated DC with the spleen lymphocytes resulted in an increased IFN-γcontent in the culture medium(32. 9 pg/ml vs 15. 1 pg/ml). Conclusion:The tumor cells with OAZI-1 overexpression can be engulfed more efficiently by macrophages and DC. And this process can induce the maturation and activation of DCs. Matured DC could induce T cell activation and then activate the anti-tumor immune response.

The expression levels of serum HSP70 and P53 in gastric cancer and its correlation with helicobacter pylori infection / 中国基层医药

Objective To study the expression levels of serum HSP70 and p53 in gastric cancer and its cor-relation with helicobacter pylori infection.Methods 50 patients with gastric cancer were selected as the research object.Among 50 cases,30 cases were stage Ⅰ -Ⅱ,20 cases were stage Ⅲ,22 cases were with lymph node metasta-sis and 28 cases were without lymph node metastasis.40 healthy subjects were selected as the healthy control group. The serum levels of HSP70 and p53 were detected.Results In the gastric cancer group,the serum levels of HSP70 and p53 were markedly higher than those of the healthy control group,and the difference was statistically significant (t =12.53,16.79,all P <0.01).Moreover,with progressing of clinical stage of gastric cancer,the serum levels of HSP70 and p53 showed a trend of increasing(t =4.68,5.29,all P <0.01).The serum expression levels of HSP70 and p53 in gastric cancer with lymph node metastasis were significantly higher than those of patients without lymph node metastasis and the difference was still statistically significant(t =3.82,4.39,all P <0.01).The serum levels of HSP70 and p53 in the gastric cancer group with Hp positive were much higher,compared to those of cases with Hp negative and the difference was also statistically significant(t =4.72,4.17,all P <0.01).Conclusion In the gastric cancer patients,the serum expression level of HSP70 and p53 are increased.The expression of HSP70 and p53 are closely related to the progress and prognosis of gastric cancer and Hp infection.

Effect of tetrabutyl propanediamine on human acute myeloid leukemia cell line HL-60 / 实用医学杂志

Objective To investigate the effect of the new polyamine analogue tetrabutyl propanediamine (TBP) on cell proliferation and the underlying mechanism. Methods MTT assay was performed to determine cell proliferation. Flow cytometry was performed to detect cell cycle transition. DNA fragmentation and mitochondrial membrane potential determinations were performed to detect cell apoptosis. The activity of key enzymes in polyamine catabolism was detected by chemiluminescence assay. Results TBP could significantly inhibit the proliferation of HL-60 cells by blockingcell cycle transition and by inducing apoptosis. The TBP-induced apoptosis of HL-60 cells was in a dose-dependent manner. The enzyme activities of SMO and APAO were also significantly increased in HL-60 cells after treatment with 100 μM TBP for 24 hours. Conclusions TBP, as a new putrescine analogue, could inhibit proliferation of HL-60 cells by increasing the enzyme activity of SMO and APAO and inducing apoptosis.

Research progress on polyamine metabolism as a target for anti-cancer therapy / 中国肿瘤临床

Rapid tumor cell growth depends on intracellular polyamine levels higher than those of normal cells. Intracellular polyamine depletion inhibits tumor cell proliferation and induces tumor cell apoptosis. Therefore, polyamine metabolism has recently been identified as an important target for anti-tumor therapy. This article briefly summarizes recent polyamine metabolism targeting, polyamine depletion within the tumor cells through a variety of methods, and the antitumor effects of the treatment.

Development of gastric precancerous lesion animal model / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a model of gastric precancerous lesion by using Aristolochic manshuriensis which contains aristolochic acids.</p><p><b>METHOD</b>The SD rats were randomly divided into four groups: control and three different doses of ethanol extractive of A. manshuriensis (EEA) (corresponding to aristolochic acid I 2.5, 5.0, 10.0 mg x kg(-1)), respectively. EEA was intragastrically given to rats every other day. At the end of the 10th, 15th, 20th week, part of the rats in each group was sacrificed and the stomachs were weighed. The gastric tumor was assessed by the weight and the relative stomach weight to the body weight. The stomachs were fixed in 4% neutral formalin, and the paraffin imbedding tissues were sliced and HE stained. Histomorphology was observed under the light microscope to determine gastric hyperplasia, mucosa precancerosis (atypical hyperplasia) and gastric cancer formation.</p><p><b>RESULT</b>The rats treated with different doses of EEA for 10 weeks induced mucosa papillary, epithelioma hyperplasia. Histological observation showed mucosa precancerosis lesions characterized as atypical hyperplasia at the dose levels corresponding to aristolochic acid I 5.0 and 10.0 mg x kg(-1) treated for 10 weeks. The incidence rate of gastric precancerosis in those two groups was 100% at the 15th week. Malignant tumors were observed in most of the animals in 10.0 mg x kg(-1) group. The animals in 5.0 mg x kg(-1) group were well tolerant compared to 10.0 mg x kg(-1) group during the course of experiment, so the dose of aristolochic acid I 5.0 mg x kg(-1) and 10-15 weeks treatment were considered to be optimum to establish the model of gastric precancerosis.</p><p><b>CONCLUSION</b>A rat model of gastric precancerosis can be induced within a short duration by giving an oral administration of the ethanol extract of A. manshuriensis which contains aristolochic acids.</p>

investigation of the anti-diabetic effects of an extract from Cladonia humilis

The effect of Cladonia humilis on glycaemic metabolism was researched in this studied. The blood glucose, insulin secretion and glycogen synthesis of the hyperglycemic mice induced by alloxan were analyzed respectively. The gluconeogenesis and the sugar tolerance of the normal mice were also analyzed in this paper. After the hyperglycemic mice were orally administrated with Cladonia humilis extract, the blood glucose was decreased [p<0.05], the level of insulin secretion and glycogen synthesis were elevated [p<0.05, p<0.01, respectively]. In addition, Cladonia humilis extract could inhibite the gluconeogenesis [p<0.01] and improve the sugar tolerance in normal control mice. These results maybe account for the causes of Cladonia humilis extract-induced significant decreases of the blood glucose in hyperglycemic mice

Development of animal model for anaphylactoid test of rodent / 中国中药杂志

<p><b>OBJECTIVE</b>To establish a simple and feasible method of anaphylactoid test on awaked small animals for screening and assessing anaphylactoid reaction of traditional Chinese medicine (TCM) injection with different concentration of tween 80.</p><p><b>METHOD</b>Test substances containing 0.4% Evans blue were intravenously injected into mice at volume of 20 mL x kg(-1) or guinea pigs at a volume of 30 mL x kg(-1). The behaviors were observed and the vascular permeability of ears evaluated by the extent of ear blue staining and absorbance of Evans blue extraction of ears were tested at 30 min after injection.</p><p><b>RESULT</b>Tween 80 solution, Yuxingcao injection with tween 80, and Shuanghuanglian powder injection obviously increased vascular permeability of ears characterized as ear blue staining and increased absorbance of the Evans blue extract from ears extracted by acetone saline both in mice and in guinea pigs in a concentration-dependent (in the case of tween 80) or a dose-dependent (Shuanghuanglian) manner.</p><p><b>CONCLUSION</b>Ear vascular permeability test in mice and guinea pigs can be used as animal models to screen and test anaphylactoid reaction induced by injections.</p>

Evaluation of vitro hepatotoxicity of monocrotaline by precision-cut liver slice technique / 中国中药杂志

<p><b>OBJECTIVE</b>To modify the empirical method of precision-cut liver slice technique, and study the hepatotoxicity of monocrotaline by this technique.</p><p><b>METHOD</b>Liver slices were prepared by the domestic shaking slicer. The technique of precision-cut liver slice was established by detecting MTT reduction used as the slice viability under different culture medium, thickness of slices, pH and culture temperature. After monocrotaline and liver slices co-culture for 6, 24 h, the slice viability, enzyme activity of GPT, GOT, LDH, GGT and protein concentration were detected by MTT reduction, enzyme kinetics method and BCA protein assay method, respectively.</p><p><b>RESULT</b>When the thickness of slices was 200 microm and pH of medium was 6.8, culture temperature was 37 degrees C, BPM culture medium, the viability of slices could maintain on a steady level. LDH leakage was significantly increased and protein content was obviously decreased after monocrotaline co-culture for 24 h with final concentration 0.02, 0.1 and 0.5 g x L(-1). No statistically significant difference between control group and monocrotaline 3 dose groups was observed in the slice viability and the content of GPT, GOT, LDH, GGT and protein after monocrotaline co-culture for 6 h.</p><p><b>CONCLUSION</b>The slice viability could retain 24 h in modified BPM medium surroundings; monocrotaline displayed liver toxicity in some degree after co-culture for 24 hours in 0.02, 0.1 and 0.5 g x L(-1) concentration.</p>

Review on experiment of traditional Chinese medicine treating to osteoporosis / 中国中药杂志

Searched the articles between 2000 and 2010, found out and summarized the articles with the topic on the experiment and new techniques of traditional Chinese medicine treating to osteoporosis. The preventive and therapeutic effect to osteoporosis by traditional Chinese medicine had been developed in the past 10 years. The study on standardization of experimental drugs, and the mechanism study with modern cell culture techniques should be enhanced.

Material and mechanisms for evaluation of Shuanghuanglian injection induced pseudoanaphylactoid reactions / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the substance basis and the mechanism of pseudoanaphylactoid reactions (PR) induced by Shuanghuanglian injection (SHLI).</p><p><b>METHOD</b>(1)The study of PR and the substance basis of PR of SHLI: ICR mice were divided into different test groups, the mice were intravenously injected with solutions of different concentration of SHLI, baicalin, forsythin, caffeotannic acid, positive control Compound 48/80 and normal sodium. All test substances were mixed with 0.4% Evans blue. The reaction and vascular permeability of the ears were observed and measured 30 min after SHLI injection. (2) The study of mechanisms: Mice were pretreated with an oral administration of Astemizol, intraperitoneal injection of cyclophosphamide 75 mg x kg(-1) or Compound 48/80 4 mg x kg(-1), then mice were intravenously injected with SHLI. At last, vascular permeability of the ears in pretreated groups was compared with SHLI treatment alone group.</p><p><b>RESULT</b>SHLI of 300 mg x kg(-1) and 600 mg x kg(-1) caused obvious vascular hyperpermeability, but baicalin, forsythin and caffeotannic didn't cause vascular hyperpermeability in the ears. The Astemizol can decrease the degree of SHLI-induced vascular hyperpermeability of the ears in the mice. After intraperitoneal injected with cyclophosphamide, there was a slight decrease in the degree of SHLI-induced vascular hyperpermeability, but there was no marked changes in the degree of the SHLI-induced vascular hyperpermeability after the mice were pretreated with Compound 48/80.</p><p><b>CONCLUSION</b>SHLI in clinic equivalent dose can cause vascular hyperpermeability. Baicalin, forsythin and caffeotannic may not result in the PR of SHLI. The mechanism of the PR maybe relate to that SHLI stimulates histamine release, the activation of leucocyte maybe take part in the SHLI-induced PR, too. Antihistamine drug can prevent the genesis of PR which induced by SHLI.</p>

Content detection of bacterial endotoxin in two kinds of injection by gelatin technique / 中国中药杂志

<p><b>OBJECTIVE</b>To detect content of bacterial endotoxin in Yuxingcao and Qingkailing injections by specific and nonspecific tachypleus amebocyte lysate technique for in order to investigate the feasibility of specific tachypleus amebocyte lysate technique for detecting bacterial endotoxin in traditional Chinese drug injections.</p><p><b>METHOD</b>Different batches of Yuxingcao and Qingkailing injections were detected by specific and nonspecific tachypleus amebocyte lysate kits.</p><p><b>RESULT</b>Yuxingcao injection could be detected by specific and nonspecific tachypleus amebocyte lysate technique, Whereas Qingkailing injection could be detected only by specific tachypleus amebocyte lysate.</p><p><b>CONCLUSION</b>Using specific tachypleus amebocyte lysate as a substitute for nonspecific tachypleus amebocyte lysate is an effective method for detecting content of bacterial endotoxin in Qingkailing injection.</p>

Detection of bacterial endotoxin content in eight kinds of injection by cytokine revulsion / 中国中药杂志

<p><b>OBJECTIVE</b>By using RAW 264.7 macrophage cell line, we studied the dose-effect relationship of endotoxin induced RAW 264.7 cells to release TNF-alpha, and then detected the content of endotoxin in 8 kinds of injections, so that we can investigate the feasibility and the interference factors of the novel test.</p><p><b>METHOD</b>By using endotoxin of different concentrations to induce RAW 264. 7 cells to release TNF-a, we drew the curve of dose-effect relationship between endotoxin and generated TNF-alpha. Then we detected the content of TNF-alpha in yuxingcao, shuanghuanglian, qingkailing, gegensu, xiangdan, qianrongmei and jiangxianmei injections and shuanghuanglian powder injection, and calculated their content of endotoxin.</p><p><b>RESULT</b>The endotoxin could induce the cells to release TNF-alpha in a good dose-dependent manner, even at a very low concentration. In the range of maximum available dilution multiple, the content of endotoxin in the rest 7 kinds of injections was less than 1.0 EU x mL(-1) except qingkailing injection of two batch.</p><p><b>CONCLUSION</b>Cytokine revulsion has the advantage of wide detection range, high sensitivity, simple operation, and the detected endotoxin is of bioactivity. This method provides another technical mean for pyrogen test of injections.</p>

Pseudoanaphylactoid reaction analysis of Chinese herbal injections in Beagle dogs / 中国中药杂志

<p><b>OBJECTIVE</b>To investigate the preclinical evaluation method of pseudoanaphylactoid reactions for Chinese herbal injections.</p><p><b>METHOD</b>Beagle dogs were divided into control group (C), 0.5% tween 80 group (T), Yuxincao injection containing 0.5% tween 80 (YT), distilled solution from Yuxincao (Y). Various groups of Beagle dogs were given 3 mL x kg(-1) of the test articles intravenously. The anaphylactoid reactions were observed immediately, while blood pressure, respiratory frequencies and heart rates were tested at 10 min and 30 min after administration.</p><p><b>RESULT</b>A variety of symptoms that range from cutaneous and mucosa signs to bronchospasma and cardiovascular collapse, including angioedema at lip, conjunctiva, ear and circumoral skin, somnolence, lethargy, breathless or dyspnea, severe hypotension etc were observed in T and TY groups from immediately post-injection to at least 30 min after administration. These reactions occurred at both first injection or repeated injections at 24 weeks intervals, manifesting that it was pseudoanaphylactoid reaction mediated by non-immune mechanisms.</p><p><b>CONCLUSION</b>Beagle dogs could be used as an animal model for preclinical evaluation of pseudoanaphylactoid reactions of Chinese herbal injection with sensitivity, reproducibility, and high clinic consistency.</p>