Objective To determine the effect of tea polyphenols on oxidative damage and (apoptosis) in human bronchial epithelial cells induced by low-dose cigarette smoke condensate (CSC).Methods We prepared CSC. 3-(4,5-dimethyl thiazoly) 2,5-diphenyl-tetrazoliun bromide (MTT) assay was used to determine the growth of cultured human bronchial epithelial cells (HBE135-E6E7). Fluorescent-chemiluminescent analyzer was used to measure cell reactive oxygen species (ROS) level. DNA ladder method was used to detect HBE135-E6E7 apoptosis. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect Bcl-2 and Bax mRNA expression.Results Concentration of intracellular ROS in the CSC group and CSC + TP group was significantly higher than that in the control group (P＜0.01); concentration of intracellular ROS in the CSC + TP group was significantly lower than that in the CSC group (P＜0.01). Apparent DNA breakage of the tail belt appeared in the CSC Group,while only a small amount of DNA breakage of the tail belt appeared in the CSC + TP group. Compared with the control group, Bcl-2 mRNA expression was reduced and Bax mRNA expression was increased in the CSC group (all P＜0.01). Compared with the CSC group, Bcl-2 mRNA expression was increased and Bax mRNA expression was reduced in the CSC+TP group (all P＜0.01). Ratio of Bcl-2 mRNA/ Bax mRNA in the CSC group and CSC+TP group was significantly lower than that in the control group (all P＜0.01).Conclusion TP can antagonize CSC-induced airway epithelial cell apoptosis through the effective removal of ROS, promoting Bcl-2 mRNA expression and inhibiting the expression of Bax mRNA.

Aim To observe the effect of polydatin(PD) on the activities of ?-AR in cardiomyocytes under the stimulation of LPS. Methods Cardiac myocytes were isolated and cultured from neonatal rats. Radioligand binding assay and flow cytometry were used to detect the function of ?-AR. Results In control group, the density of ?-AR (B max) was 4.14?1.41 fmol?(1?105 cells) -1 and ?-AR affinity (K d) was (4.02?0.59) nmol?L -1. In LPS group, ?-AR density was down-regulated 〔B max=1.78?0.12 fmol?(1?105 cells) -1〕 and the affinity decreased (K d=23.88?2.34 nmol?L -1). In LPS+PD group, ?-AR B max was recovered to 3.37?0.36 fmol?(1?105 cells) -1 and the affinity increased (K d=3.44?0.44 nmol?L -1). In PD group, ?-AR functions had no significant difference with normal group 〔B max=2.76?0.32 fmol?(1?105 cells) -1 and K d=4.63?1.54 nmol?L -1〕. The ?-AR amount measured by flow cytometry showed the same change tendency as the radioligand binding assay. Conclusion ?-AR activities may be down-regulated during the pathological process of LPS treatment. PD could reverse the effects of LPS by up-regulating ?-adrenoreceptors.

AIM: To investigate the chemotaxis mechanisms of polymerphonuclear leukocytes (PMNs) under the stimulation of LPS and its regulation by polydatin (PD). METHODS: Chemotactic chamber assay was used to investigate the regulatory role of formyl peptide receptor like-1 (FPRL1) in the chemotaxis of PMNs in response to LPS and PD. RESULTS: LPS increased the secretion of PMN chemotactic factor and up-regulated the function of formyl peptide receptor like-1. PD did not obviously affect the chemotactic factor secretion in normal PMN and the function of formyl peptide receptor like-1, but it significantly reduced the rise of chemotactic factor excretion in PMNs caused by LPS stimulation and PMNs transfected with formyl peptide receptor like-1. CONCLUSION: FPRL1 may mediate LPS-induced PMN chemotaxis, and PD regulates PMN chemotaxis by down-regulating the secretion of chemokine and the function of FPRL1 and may play a crucial role in the treatment of inflammation.

OBJECTIVE:To optimize?-cyclodextrin inclusion process in the inclusion of volatile oil compounds of peppermint,biond magnolia flower and agastache rugosa in biyuanling granules.METHODS:Orthogonal experiment was performed,the inclusion process was evaluated with inclusion rate of volatile oil,recovery rate of inclusion compound,extract ratio of inclusion compound as indexes.RESULTS:The optimized inclusion condition was as follows,the ratio of volatile oil to?-cyclodextrin was1∶8(ml/g)with10min ultrasound time and20℃ultrasound water temperature.CONCLUSIONS:The optimized method was simple and efficient,and suitable for industrialized production.

BACKGROUND:Now in mammary stem cellresearch, no proper mammary stem cellmedium is provided to culture mammary stem cells. OBJECTIVE:To create a mammary stem cellmedium and validate its application by isolating Sca-1+mammary stem cells. METHODS:We first used BM medium to culture mammary organoids, and after 6 days, the expression of Sca-1 and vimentin was detected in fibroblasts by immunofluoresence method. Then, we established MaECM medium which arrested fibroblasts growth. After 6 days culture of mammary organoids by MaECM medium, Sca-1+and Sca-1-cellpopulations were sorted out by magnetic sorting and the purity was analyzed by flow cytometry. Sorted 1×104 Sca-1+or Sca-1-cells were transplanted into the bilateral mammary fat pads of four mice, and after 6-8 weeks, the fat pads were harvested for whole-mount immunohistochemical analysis and hematoxylin-eosin staining. RESULTS AND CONCLUSION:After 6 days culture of mammary organoids under BM medium, smal-sized colonies were generated around lots of fibroblasts. Immunofluoresence staining detected strong expression of vimentin and Sca-1 in fibroblasts, indicating that the BM medium is not suitable to isolate Sca-1+mammary stem cell. The MaECM medium promoted the proliferation of mammary epithelial cells whereas arrested fibroblasts growth. After 6 days culture of mammary organoids under MaECM medium and magnetic sorting, the flow cytometry showed that the purity of Sca-1+cellreached 92%and 5%in the Sca-1+and Sca-1-population, respectively. The results from transplantation test showed that six mammary outgrowths were regenerated out of eight injected fat pads in the Sca-1+cells transplantation, but in the Sca-1-transplantation population, one mouse died and the other transplants failed to produce outgrowths. We developed the MaECM medium which promoted the proliferation of mammary epithelial cells whereas arrested Sca-1+fibroblasts growth. Using the medium, we confirmed that Sca-1+mammary cells have capacity of isolating mammary stem cells.

Objective To observe the effects of Jieyu-Chufan capsule(JCC) on the behavior and the monoamine neurotransmitter content of cerebral in depression mouse model and explore its mechanism. Methods The ICR male mouse were randomly divided into seven groups according to their weights(the model group, the low, middle and high doses group, Baiyoujie group, the amitriptyline group, the normal control group). The low, middle and high doses group were given JCC with 1.250, 2.500, 5.000 g/kg separately by intragastric administration. Baiyoujie group was given 0.014 g/kg Baiyoujie by gavage. The amitriptyline group was given 0.050 g/kg amitriptyline by gavage. The model group and the normal control group received the same volume of distilled water intragastrically. The drugs were administrated once a day. The brepharoptosis and movement conditions of mouse were observed after 22 days. The monoamine neurotransmitter content of each group was measured after 25 days. Results Compared to the model group, the number of mouse with brepharoptosis decreased in the high dose group after 1, 2, 6 hour with reserpine injection (P<0.05 or P<0.01);the number of mouse came out from the circle increased after 4 hours with reserpine injection (P<0.05 or P<0.01);the content of NA (880.45 ± 428.81 ng/g, 875.98 ± 449.33 ng/g vs. 299.92 ± 267.08 ng/g) and DA (2 305.99 ± 530.37 ng/g, 2 169.99 ± 278.19 ng/g vs. 1 439.34 ± 357.33 ng/g) in the middle and high dose group increased (P<0.01);the content of 5-HT (781.43 ± 135.10 ng/g vs. 492.01 ± 192.80 ng/g) in the middle group increased (P<0.01). Conclusions JCC showed an antidepression influence in depression mouse model induced by reserpine. The mechanism may be related to regulating the level of monoamine neurotransmitter in the nervous centralis .

Aim To study the influence of PD on the activity of PKC(protein kinase C) of VSMC during ischemia and hypoxia.Methods The hypoxia model was made by drawing off the oxygen of an enclosed acryl glass box and ischemia model was made by low sugar culture medium. Then the activity of PKC was measured by ?-scintillation counting instrument.Results It was found that the activity of PKC of cytoplasm in VSMC in ischemia and hypoxia situation increased (vs normal group P0.05 ).Conclusion PD can reverse the PKC activities induced by ischemia and hypoxia. It may be one of the mechanisms of PD to have antishock effect.

OBJECTIVE: To study the optimum technique for extraction of mixture volatile oil from Olibanam and Sandalwood. METHODS: The orthogonal test was adopted to optimize the extraction process using the yield rate of volatile oil as indicator. RESULTS: The optimum extraction conditions were as follows: powder forming by 10mesh, 10- fold water added, soaking for 10h and extraction for 8h. CONCLUSIONS: This extraction method is simple, efficient and suitable for industria-lized production.

To investigate the knowledge, attitude and practice (KAP) on rabies among a mass of the exposed population in Beijing so as to provide scientific evidence for development of measures on the prevention and protection of rabies, the descriptive studies were employed, in which patients were interviewed face to face with a standard questionnaire in hospital or the center for disease control and prevention (CDC), including history of exposure, knowledge on rabies, post-exposure treatment etc. Of 478 respondents, 76% of them were bitten on the upper limbs with the exposure proportions in category Ⅰ, Ⅱ and Ⅲ of 24%, 70% and 6% respectively. The proportions of awareness of knowledge in rabies before biting, its prevention, the correct measure to treat their wounds themselves, rabies vaccine inoculation, and injections with rabies antiserum constituted in 82%, 55%, 59%, 99% and 22% respectively. Result of this investigation indicates that most of the people investigated had a lower awareness of knowledge on rabies, the correct measures to treat rabies after biting as well as the measures to prevent rabies, suggesting that community-based prevention and control measures should be emphasized, especially targeting the population with high risk and the heath education on rabies should performed promptly by several governmental departments.

Objective To investigate the effect of serum of esophageal squamous cell carcinoma (ESCC) patients with blood stasis syndrome (BSS) on proliferation and cycle of EC9706 cells, and to explore the action of blood micro-environment of ESCC patient with BSS on EC9706 cells. Methods Human EC9706 cells were cultured in an incubator with RPMI-1640 medium containing fetal bovine serum ( FBS) , at 37℃ and under 5% saturated humidity for 24 h. After EC9706 cells were starved in serum-free medium for another 24h, the three experimental groups were treated with serum of ESCC patients with BSS, serum of ESCC patients with spleen-qi deficiency syndrome (SQDS), and serum from healthy volunteers, respectively. Cell proliferation was determined by methyl thiazolyl tetrazolium ( MTT) assay, EC9706 cell morphology was observed under light microscope, and cell cycle was measured by flow cytometer (FCM). Results The serum concentrations of ESCC patients with BSS and ESCC patients with SQDS for obtaining 50 percent cell proliferation rates ( PI50) were 71.1 μL/mL and 118 μL/mL, respectively. And the proliferation of EC9706 cells in the both groups all arrived to the peak values at culturing hour 48. The light microscopy results showed that the feature of EC9706 cells in both groups presented as spindle-like or polygon-like shape, and cell count in BSS group was larger than SQDS group. FCM assay results for EC9706 cell cycle showed that the percentage of G1-phase EC9706 was decreased and the percentage of S-phase EC9706 was increased in BSS group as compared with those in SQDS group ( P<0.05). Conclusion Serum micro -environment in ESCC patients with BSS is more beneficial to EC9706 cells proliferation than ESCC patients with SQDS, and the mechanism may be related to the regulation of cell cycle.