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Objective:To explore the proliferation inhibition and radiosensitization of Biyanqing granule on nasopharyngeal carci -noma cell line CNE-2 in vitro.Methods:CNE-2 cells were cultured in vitro.The inhibition of Biyanqing granule on the proliferation of CNE-2 cell was evaluated by MTT assay .Radiosensitization was explored by clone formation assay , and cell cycle and apopotosis were observed by flow cytometry ( FCM) .Results:Biyanqing granule could inhibit the proliferation of CNE-2 in a time-and dose-dependent manner.The IC50 in 24, 48 and 72 h was 70.79, 60.13 and 51.63 mg· ml-1(calculated according to the weight of all medicinal ma-terial), respectively.The colony formation assay showed that Biyanqing granule combined with radiation could significantly reduce the colony formation of CNE-2 cells.With the concentration increase of the main drug , the colony formation of CNE-2 cells was reduced . The number of colony formation in the negative control group , the radiation group , 10 mg· ml-1 and 20 mg· ml-1 Biyanqing combined with radiation groups (calculated according to the weight of all medicinal material ) was significant different (P<0.05).With the main drug concentration increasing , the percentage of G 2/M phase and apoptotic cells were both increased , and compared with the con-trol group, the difference was significant (P<0.05).Conclusion:Biyanqing granule can not only inhibit CNE-2 cells but also block CNE-2 cells in G2/M to improve the radiosensitization of CNE-2 cells.
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Objective To construct a prokaryotic expression plasmid for CPSIT_p7 gene from Chlamydophila psittaci ( Cps) 6BC strain and to evaluate immunogenicity of the recombinant protein His-CPSIT_p7 and detect its dynamic expression at mRNA and protein levels in HeLa cells during persistent Cps infection.Methods The fusion protein His-CPSIT_p7 was expressed in E.coli BL21 and purified by Ni-NTA affinity chromatography .BALB/c mice were immunized with the recombinant protein to prepare polyclonal antibody for evaluation of the immunogenicity of His-CPSIT_p7 by ELISA.Penicillin sodium was used to establish a model of Cps persistence infection .RT-PCR and Western blot assay were performed to de-tect the expression of CPSIT_p7 at mRNA and protein levels during Cps persistent infection .Results The fusion protein His-CPSIT_p7 was successfully expressed with the use of constructed recombinant expression plasmid pET30 a-CPSIT_p7 and purified .ELISA result showed that the specific antibody titer against CPSIT_p7 reached 1 ∶1 000 000 on the 40th days after immunization .The expression of CPSIT_p7 at mRNA and protein levels were increased in a time-dependent manner in Cps-infected HeLa cells .The peak of mRNA level was reached at the time point of 36 hours after infection , followed by a time-dependent decrease during Cps acute infection .However , the expression of CPSIT_p7 at mRNA and protein levels were not decreased until 60 hours after infection during Cps persistent infection .Conclusion His-CPSIT_p7 protein was suc-cessfully expressed in the prokaryotic expression system and purified , showing an advantage of good immuno-genicity.Highly expressed CPSIT_p7 at mRNA and protein levels were detected during Cps persistent infection.
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BACKGROUND: Qiangji jianli liquid plays a key role in prevention and cure of myasthenia gravis; however, whether changes of nucleic acid and protein are related to its mechanism or not should be studied further.OBJECTIVE: To observe the effect of qiangji jianli liquid on synthesis of nucleic acid and protein in mice with experimental spleen deficiency syndrome, and investigate the effect on myasthenia gravis and the molecular biological basis.DESIGN: Randomized controlled animal study.SETTING: Testing Center and Basic Medical College of Guangzhou University of Traditional Chinese Medicine.MATERIALS: Healthy male NIH mice, aged 4 weeks, of clean grade, weighing 17-21 g, were provided by Medical Experimental Animal Center of Guangdong Province. Qiangji jianli liquid (Guangdong Dongfang Shencao Pharmaceutical Factory); serpate solution (Guangdong Bangmin Pharmaceutical Factory); [5-methyl-3H] TdR, [5-3H] uridine and L-[4,5-3H]leucine (Atom High-nuclear Technology Application Limited Company).METHODS: The experiment was carried out in the Laboratory of Nuclear Medicine of Guangzhou University of Traditional Chinese Medicine from June to November 2005. A total of 180 healthy male NIH mice were randomly divided into normal control group, model group, 26 g/kg qiangji jianli liquid group, 13 g/kg qiangjijianli liquid group and sijunzi decoction group with 36 in each group. Except normal control group, serpate was used to establish animal models of spleen deficiency syndrome. 0.2 mg/(kg ·d) serpate was intraperitoneally injected into mice. ① 0.1 mL/(mouse·d) saline was intraperitoneally injected into mice in normal control group. ② 26 g/kg and 13 g/kg were perfused into mice in 26 g/kg and 13 g/kg qiangji jianli liquid groups, respectively, once a day. Qiangji jianli liquid consisted of beiqi, dangshen, shengma,baizhu, gaicao, etc., with 1.204 g raw drugs a milliliter. ③ 26 g/kg sijunzi decoction, which consisted of dangshen,baizhu, fuling, jiugancao, etc., was perfused into mice with the dosage of 0.5 mL/mouse in si junzi decoction group once a day. After 16 successive days, contents of DNA, RNA and protein were measured in spleen and kidney tissue; meanwhile, body mass, ratio between spleen mass and body mass, and ratio between kidney mass and body mass were also measured before modeling and after administration.MAIN OUTCOME MEASURES: ① Changes of body mass before modeling and at 16 days after administration; ② changes of mass of spleen and kidney tissues; ③ Comparisons of DNA, RNA and protein in spleen and kidney tissues at 16 days after administration.RESULTS: All 180 mice were involved in the final analysis without any loss. ① There was no significant difference of body mass (17.4-21 g) among groups before experiment (P =0.993); however, after experiment, body mass in model group was lower than that in normal control group (Q =22.84, P ＜ 0.01); 16 days after administration, body mass was higher in qiangji jianli liquid groups (26 g/kg, 13 g/kg) and sijunzi decoction group than that in model group (Q =8.66-10.24, P ＜ 0.01). ② Values of spleen mass/body mass and kidney mass/body mass were decreased in model group as compared with those in normal control group (Q =4.02-12.93, P ＜ 0.01); masses of spleen and kidney tissues were increased in qiangji jianli liquid groups (26 g/kg, 13 g/kg) and sijunzi decoction group as compared with those in model group (Q =3.21-9.69, P ＜ 0.05-0.01); ratios in 13 g/kg qiangji jianli liquid group were higher than those in model group (Q =4.07, 5.92; P ＜ 0.05, 0.01). ③ Contents of DNA, RNA and protein in spleen tissue and contents of RNA and protein in kidney tissue were lower in model group than those in normal control group (Q =7.15-19.2, P＜ 0.01); however, content of DNA in kidney tissue was higher in model group than that in normal control group (Q =4.19, P ＜ 0.05). Contents of DNA, RNA and protein in spleen tissue and contents of RNA and protein in kidney tissue were higher in 26 g/kg qiangji jianli liquid group and sijunzi decoction group than those in model group (Q =2.91-14.12, P ＜ 0.05-0.01 ).CONCLUSION: Qiangji jianli liquid can accelerate synthesis of nucleic acid and protein; additionally, onset of spleen deficiency syndrome may be related to decreasing synthesis of nucleic acid and protein.
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【Objective】To observe the morphologic changes of organs in mice with spleen deficiency and the effect of Qiangji Jianli Oral Liquid(QJOL).【Methods】Forty healthy male NIH mice were randomized into 4 groups:A(normal),B(model),C(treated with QJOL)and D(treated with Si Junzi Decoction).Except group A,the mice in other 3 groups received intraperitoneal injection of reserpine to induce spleen deficiency.Meanwhile,groups C and D received QJOL and Si Junzi Decoction in the dosage of 26?g?kg~(-1)?d~(-1) by gastric gavage respectively,and groups A and B received the same volume of distilled water for 21 days.Twelve hours after last administration,the body weight and pathological changes of liver,kidney,spleen and thymus were examined.【Results】In group A,the body weight decreased,and liver,kidney,spleen and thymus gland showed different degrees of damage.QJOL and Si Junzi Decoction could increase the body weight and promote the repair of the organs.【Conclusion】Different degrees of damage can be found in liver,kidney,spleen and thymus gland of mice with spleen deficiency,and spleen-strengthening medicinal herbs can promote their repair,indicating that the theory of spleen controlling the muscles,being the solitude organ and localized the center to cultivate the surroundings organs like the earth,which is described in Huangdi's Canon of Medicine,has the morphologic evidence.
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[Objective] To explore the relationship between neuroendocrine changes caused by melatonin (MT) hypo-secretion and kidney-in deficiency syvndrome during prepubescence. [Methods] Thirty SD rats after ablactation were randomly divided into normalgroup (group. A), model group (group B) and herbal medicine group (group C, treated with kidney-yin-nourishing ande(?)ficiency-fire-clearing herbs). Rats in groups B and C were exposed to light continuously for 30 days and simultaneously groupC was given kidney-yin-nourishing and deficiency-fire-clearing herbal medicine (13.3 g?kg-1?d-1, qd) for 30 days . After treatment, blood samples were collected at 0:00-2:00 o'clock to observe the changes of blood MT, testosterone (T), epinephrine (E) and norepinephrine (NE) levels. [Results] Body weight and blood MT level decreased, and T, NE and E levels increased in group A (P
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[Objective] To observe levels of plasma endothelin (ET) and calcitonin gene - related peptide (CGRP) in rats with myocardial hypertrophic heart failure affected by Yang Xin Kang (composed of Radix Ginseng, Radix Ophio-pogonis, Radix flicis Pubescetis, etc.) and its mechanism. [Methods] Rat models with myocardial hypertrophic heart failure was established by subcutaneous injection of isoprenaline for 13 days. Fifty rats were allocated to five groups: normal control (Group A), model (Group B), digoxin (Group C), high dosage of Yang Xin Kang (Group D) and low dosage of Yang Xin Kang (Group E). ET and CGRP levels were measured by radioimmunoassay method. [Results] ET level in Group B was higher and CGRP level lower than those in Group A (P
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Objective The changes of thyroid hormone and cyclic nucleotides levels in rats with spleen-kidney deficiency syndrome were observed to evaluate the preventive and therapeutic effect of Yougui Wan(YW) for spleen-kidney deficiency syndrome,and to explore the relationship between the spleen and the kidney at microscopic level.Methods Male SD rats were randomly divided into normal control group,model group,and YW group.Rat models of spleen-kidney deficiency syndrome were established by gavage firstly with rhubarb(4g for each rat) decoction and then with hydrocortisone(25 mg?kg-1?d-1,im).The model group and YW group were treated with distilled water and YW(5.8 g?kg-1?d-1) respectively.The general status of rats was observed,and the plasma cyclic adenosine monophosphate(cAMP) and cyclic guanosine monophosphate(cGMP) levels as well as serum thyroid hormones(T3,T4 and rT3) and thyroid-stimulating hormone(TSH) contents were determined in all rats by radioimmunoassay(RIA).Results Compared with the normal control group,the serum contents of T3,T4,rT3 were decreased significantly(P
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【Objective】To observe the effect of Buzhong Yiqi Wan(BYW)on thyroid hormones in rat model of spleen-asthenia.【Methods】Thirty-four SD male rats were randomized into 3 groups:normal control group(N=10),spleen-asthenia model group(N=12),BYW group(N=12).Except the normal control group,the rats in other groups were given the decoction of Radix et Rhizoma Rhei(4 g?d-1 for each rat)to induce spleen asthenia,and meanwhile,BYW group reveived BYW(3.9 g?kg-1?d-1).The treatment lasted 20 days.Radioimmunoassay was used to detect serum levels of triiodothyronine(T3),thyroxine(T4),reverse triiodothyronine(rT3)and thyroid stimulating hormone(TSH).Meanwhile,the body weight,spleen and thymus weight as well as their ratio with the body weight were also observed.【Results】Serum T3 and T4 levels,spleen and thymus weight as well as their ratio with the body weight were decreased in the model group as compared with those in the normal control group(P
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[Objective] To investigate the effect of Bao Xin Kang (BXK) on levels of cyclic adenosine monophosphate (cGMP) and cyclic guanosine monophosphate (cAMP) in vascular smooth muscle of rats with myocatdial hypertrophic heart failure and to explore its mechanism. [Methods] Fifty rats were randomly allocated to five groups: normal control group (Group A) , model group (Group B) , digoxin group (Group C), large - dose BXK group (Group D) and small - dose BXK group (Group E). Rat models with myocardial hypertrophic heart failure were established by subcutaneous injection of isoprenaline for 13 days. Radioimmunoassay was used to measure the levels of cAMP, cGMP and the ratio of cAMP/cGMP in vascular smooth muscle. The body weight and the weight of heart, liver and lung were measured and his-tomorphologic features of heart, liver and lung also examined. [Results] The levels of cAMP and cGMP and the heart index (heart weight /body weight) and the liver index (liver weight/body weight) in Group B were increased as compared with those in Group A (P
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Objective To explore the mechanism of Shema Zhichuan Liquid (SZL) for asthma. Methods Forty rats were allocated to normal control group (Group A), model group (Group B),western medicine group (Group C) and SZL group (Group D). Radioimmunoassay was used to observe the levels of cyclic adenosine monophosphate (cAMP) and cyclic guanosine monophosphate (cGMP) and the ratio of cAMP/cGMP.Results After treatment, cAMP level and cAMP/cGMP ratio were increased and cGMP level decereased in Group C and Group D as compared with Group B (P
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Objective To study effects of baicalin(Bai) on AngⅡ in diabetic nephropathy(DN) rats and to explore its preventive and therapeutic effect on treating DN from the non-hemodynamic mechanism of AngⅡ.Methods Nineteen specific pathogen-free male rats were randomly divided into three groups:normal control group(5 rats),diabetic nephropathy model control group(7 rats),baicalin treated group(7 rats).DN rat model was established by one-dose intraperitoneal injection of streptozotocin.Seven weeks after the modeling,rats in baicalin group recieved intraperitoneal injection of baicalin solution(40 mg/kg),rats in DN model group received intraperitoneal injection of normal saline(1 mL for each rats),and the normal control group did not receive any intraperitoneal injection.After treatment for 6 weeks,orbital blood from all rats was collected to determine plasma AngⅡ by radioimmunoassay.Unilateral kidney was processed by 4 % paraformaldehyde fixation and then was used for the determination of TGF-? by immunohistochemical assay.The other unilateral kidney was got out to prepare for renal cortex homogenate,and then was used for the determination of renal tissue AngⅡ by radioimmuno assay.Results The quantity and degree of TGF-? expression in baicalin group was decreased,and there was statistical significance between baicalin group and the model group(P
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Objective To investigate the effects of Huangqi Sijun Decoction on the levels of thyroxin and cAMP and cGMP in model rats with spleen deficiency syndrome and to explore its therapeutic mechanism.Methods Male SD rats aged three months were randomly divided into 3 groups:normal control group,spleen deficiency model group and Huangqi Sijun Decoction group.The rat models of spleen deficiency syndrome were established by gavage with the decoction of Rhubarb.The serum levels of thyroxin(T4),3,5,3'-triiodothyronine(T3),thyrotropin(TSH)and plasma levels of cAMP,cGMP were determined.Results T3 and T4 were significantly lower but the levels of cAMP and cAMP/cGMP were significantly higher in the spleen deficiency models than those in the normal control group(P
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SD male rats aged 21 days were continuously exposed to light for 30 days in order to inhibit secretion of melatonin (MT). Changes in serum MT, testosterone, corticosterone, thyroid hormones (T_ 3 , T_ 4 ), adrenalin and noradrenalin were determined. The results showed that MT inhibited the functions of gonad, adrenal and sympathetic nerve, but increased the function of thyroid in pre-puberty rats.