Objective We established a model of human breast cancer in nude mice, to discuss the feature of pathology and biology of breast cancer,and give help to establish tools of pathogen research. Methods Human breast cancer cells MCF-7 were subcutaneously injected into the right armpit of nude mice to establish human breast cancer models.The mice were divided into composite group, estrogen group, leptin group and the blank group (30 in each). In the composite group,estrogen and leptin were injected into peripheral region of the tumor daily.In the estrogen group,estrogen was injected.In the leptin group, leptin was injected.In the blank group, physiological saline was injected.Tumor growth was observed, and the volume of the tumor was recorded.The tumor tissues obtained from the mice were pathologically examined. Results (1)The tumor-taking rate of leptin group and the blank group were 46.7% (14/30)and33.3% (10/30) ,and the mode is failure.In composite group and estrogen group they were 96.7% (29/30) and 70% (28/30).There was not significant difference between them ( P < 0.05).(2) The differences of average diameter and volume were statistically significant between the composite group and the estrogen group (P < 0.05).(3) Pathology diagnosis was invasive ductal carcinoma. Conclusions (1) Establishing human breast cancer model in nude mice need the stimulation of estrogen. The tumor-taking rate of nude mice has no relationship with leptin.(2) In the study in vivo, leptin as same as estrogen has stimulating effect on MCF-7 cells proliferation.(3) The transplanted cancer cell partly have the pathology and biology features of the human breast cancer cells and give help to establish tools of pathogen research.

Objective To compare the efficacy and safety of topical anesthesia,conscious sedation and intravenous anesthesia during endoscopic varices ligation(EVL).Methods Patients underwent EVL were divided into 3 groups to receive different anesthetic methods,namely topical anesthesia,conscious sedation and intravenous anesthesia,with 50 subjects in each group.The changes of vital signs,the tolerance to stimulation of the procedure,the time of operation,the rate of complication were recorded and compared between 3 groups.Results The procedure of EVL were completed in all patients.In topical anesthesia group,40(80%)patients had nausea and vomiting,9 cases(1 8%)tried to pull out the endoscopy.The mucosa at the entrance of the esophagus were injured in 10 cases(20%).Massive bleeding occurred in 4 patients(8%)during operation because of nausea and vomiting.In conscious sedation group,only 7 patients(14%)had mild nausea and vomiting,and no complication of variceal bleeding occurred.The mucosa at the entrance of the esophagus was injured in 5 cases(10%).In intravenous anesthesia group,no patient had nausea or vomiting.The respiratory rate,heart rate and mean artery pressure decreased during the procedure,but without significant difference(P＞0.05).The operation time in intravenous anesthesia group WaS shorter than that in other two groups,but without significant difference(P＞0.05).Conclusion EVL can be completed under 3 different anesthetic methods,while EVL under conscious sedation is more effective and safe.

ObjectiveTo investigate the inhibitory effect of lentivirusly-mediated ObR-siRNA on transplanted MCF-7 human breast cancer cells by intratumoral injection.MethodsA model of subcutaneous implanted tumor was generated through injecting MCF-7 human breast cancer cells into the nude mice.Thirty established mice with MCF-7 breast cancer cells xenograft were divided into 3 groups randomly,and mice in the experimental group were intratumorally injected with ObR-siRNA lentivirus,while the negative control group and blank control group mice were injected with the same dose of negative lentivirus and normal saline.All mice were subcutaneously injected with recombinant human leptin around the tumor site once a day.Tumor size was blindly measured every other day and the mRNA expression and protein expression levels of ObR in each group were determined.ResultsKnockdown of ObR-treated xenografted nude mice with a high leptin microenvironment was successfully established.Local injection of ObR-siRNA lentivirus significantly suppressed the established tumor growth in nude mice(P ＜ 0.01,P ＜0.01 ).Real time-PCR and Western blotting showed that the mRNA and protein expression of ObR was decreased in the ObR-siRNA lentivirus group( P ＜ 0.01,P ＜ 0.01 ).ConclusionsIntratumoral injection of recomhinant ObR-siRNA lentivirus inhibits the growth of MCF-7 cells xenografts in the nude mice,suggesting that ObR might represent a therapeutic target in the genotherapies of human breast cancer.

Objective To observe the changes on the neurogenic inflammation-related factors in the dura mater of the rat model of migraine and investigate the possible mechanism of the pain of migraine.Methods SD rats were randomly divided into stimulation group ( n = 32 ) and sham group ( n = 32 ).Unilateral trigeminal ganglion was stimulated to induce migraine for rats in the stimulation group. Rats in the sham group were subjected to sham surgery. The levels of calcitonin gene-related peptide (CGRP) in the blood of jugular vein in the stimulation side were measured by radioimmunoassay. The levels of histamine in peripheral blood and prostaglandin E2 (PGE2 ) in the dura mater were determined by enzyme-linked immunosorbent assay (ELISA). The number of mast cells and percentage of their degranulation in the dura mater were determined under a microscope after toluidine blue staining. Cyclooxygenase 2 (COX-2)expression in the dura mater was evaluated by immunohistochemical staining and western blot analysis. Results In the stimulation group, the level of CGRP in the ipsilateral jugular vein was (82. 84 ± 16. 24)pg/ml and in the sham group was (59. 20 ±11.66) pg/ml (t = -3.34, P ＜ 0. 05 ). The level of histamine in the ipsilateral jugular vein was ( 11.59 ± 1.20) ng/ml and in the sham group was (9. 87 ±0. 88) ng/ml (t = - 3. 27, P ＜ 0. 05). The number of mast cells in the dura mater decreased from 15.46 ± 2. 40 in the stimulation group to 11.63 ± 1.67 in the sham group ( t = 3.71, P ＜ 0. 05 ). Degranulation of mast cells in the dura mater significantly increased from 14. 09% ±4. 53% in the sham group to 29. 10% ±9. 39% in the stimulation group (t = - 4. 07, P ＜ 0. 05 ). The level of PGE2 in the stimulation group was ( 382. 30 ±20. 90) pg/ml and in the sham group was (80. 70 ± 10. 60) pg/ml (t = - 16. 674, P ＜0. 05). The number of COX-2 positive cells significantly increased from 42. 00 ± 18.40 in the sham group to 139.00 ±20. 50 in the stimulation group (t = -7. 994, P ＜0. 05). Also the COX-2 protein level was elevated from 19. 50 ±9. 20 in the sham group to 359. 20 ±21.90 in the stimulation group (t = -5. 190, P ＜0. 05). Conclusions Electrical stimulation on the unilateral trigeminal ganglion induces neurogenic inflammation in the dura mater. Changes on the neurogenic inflammation-related factors are probably the essential pathophysiological mechanism underlying the pain in migraine.

Objective To investigate the cytotoxicity of silica nanoparticles on vascular endothelial cells, and to clarify its action mechanism.Methods The 60 nm silica nanoparticle was selected and the invitro cultured human umbilical vein endothelial cells (HUVECs)were used as cell model.The HUVECs were divided into control and silica nanoparticle exposure groups with concentrations of 12.5,25.0,and 100.00 mg·L-1 .MTT assay was used for the determination of cell viability,lactate dehydrogenase (LDH)release assay for membrane integrity,flow cytometry (FCM)for intracellular reactive oxygen species (ROS)content,and real-time PCR assay for intracellular NF-E2-related factor 2 (Nrf2 ), heme oxygenase-1 (HO-1 ), superoxide dismutase 2 (SOD2 ) and glutamate-cysteine ligase catalytic subunit (GCLC)mRNA levels.Results The MTT results showed that the cell viabilities in each silica nnaoparticle exposure group were decreased compared with control group in a dose-dependent manner. Upon the silica nanoparticle exposure for 12 h,the cell viability was declined significantly only in 100 mg·L-1 exposure group compared with control group (P<0.05).When exposured for 24 h,the cell viabilities in 25.0, 50.0,and 100.0 mg·L-1 exposure groups were declined significantly compared with control group (P<0.05). Under the exposure to silica nanoparticle with the same dose, the cell viabilities were decreased along with the elongation of exposure time.LDH assay and FCM showed that except for that in 12.5 mg·L-1 exposure group, both the LDH activities in media and intracellular ROS levels in other exposure groups were increased compared with control group (P<0.05 ). The results of real-time fluorescence PCR showed that the mRNA levels of Nrf2, HO-1,SOD2 and GCLC in 100 mg·L-1 silica nanoparticle exposure group were increased significantly compared with control group (P<0.05).Conclusion Silica nanoparticles have toxicity to vascular endothelial cells,which includes reducing cell viability,membrane integrity destruction,induction of ROS generation,and tranSCriptional regulation of redox-related factors. Oxidative damage is one of the mechanisms of vascular endothelial toxicity mediated by silica nanoparticles.

This paper was aimed to study effects of different preparation methods on the content of ginsenosides Rg1,Re and Rb1 in Yi-Xin-Shu (YXS) tablets by HPLC-ELSD.HPLC-ELSD was used as the detection method.The separation and content of ginsenosides Rg1,Re and Rb1 were used as indexes.The influences of three different preparation methods (i.e.,defatted alcohol extraction and butanol extraction,alcohol extraction and butanol extraction,alcohol extraction and butanol extraction ammonia solution washing) on the effect of YXS tablets were studied.Then,the same content determination method was used to compare the influence of alkali washing treatment to ginsenosides Rg1,Re and Rb1 among different batches of Panax ginseng.The results showed that a good separation of ginsenosides Rg1,Re and Rb1 component peak of YXS tablets was achieved by three kinds of separation methods.The separation degree was greater than 1.5.Ammonia solution washing had some effect on ginsenosides Rg1,Re and Rb1 content,which made the content of ginsenosides Rg1,Re and Rb1 be 1.5-1.8 times to those without alkali washing.No effect was shown on the content of ginsenosides Rg1,Re and Rb1 during ammonia solution washing.It was concluded that some other ginsenosides can be transferred into ginsenosides Rg1,Re and Rb1 in YXS tablets solution after ammonia solution washing.

This study was aimed to improve the transfer rate of sinomenine in the intermediate product of Caulis Sinomenii in order to optimize the purification process of intermediate product ofCaulis Sinomenii fromTong-An (TA) injection. The transfer rate of sinomenine and the stability of fingerprints in the intermediate product of Caulis Sinomenii were used as indexes for the investigation on the impact from different pH ofCaulis Sinomenii extract before extraction. The transfer rate of sinomenine was used as an index for the investigation on the impact by different pH of hydrochloric acid created to dry extract solution. The transfer rate of sinomenine was used as an index for the investigation of four separation ways, which included the vacuum filtration, plate and frame filters, high-speed tube separator, and flat direct centrifuge, on the liquid separation of sinomenium acutum acid. The results showed that the pH ofCaulis Sinomenii extract before extraction was 10-11; the transfer rate of sinomenine was the highest in the extraction process and the fingerprints of TA injection was stable. The pH of hydrochloric acid was 2.0-2.5; and the highest transfer rate of sinomenine in acid dissolution process was 92.94%. The high-speed tube separator had the best separation to sinomenium acutum acid-dissolving liquid. The highest transfer rate of sinomenine was 93.34%. It was concluded that the optimized process can effectively improve the transfer rate of sinomenine in the intermediate product ofCaulis Sinomenii. Meanwhile, fingerprints of the product were stable. The process was simple with good repeatability.

Objective To investigate the expression of endostatin and angiopoietin (Ang)-2 in human ghoma and its significance. Methods The expression of endostatin and Ang-2 were measured by immunohistochemistry and endostatin mBNA by hybridization in situ in 108 cases of brain glioma and 5 cases of normal brain tissues. Results The expression of endostatin (0.0657±0.0038)and Ang-2 (0.0286± 0.0042) were significantly higher in grade Ⅲ-Ⅳ glioma patients than those in grade Ⅰ-Ⅱ ghoma patients (0.0349±0.0048,0.0084±0.0018, respectively) and normal brain tissues (0,0)(P<0.01). The expression of endostatin mRNA were significantly higher in grade Ⅲ-Ⅳ glioma patients (0.0310±0.0041) than that in grade Ⅰ-Ⅱ glioma patients (0.0152±0.0031) and normal brain tissues (0)(P< 0.01 ). Theratio of endo-stalin to Ang-2 was negatively rehted to the grade of glioma (r=-0.810,P <0.01). Conclusion The interaction of endostatin and Ang-2 plays an important role in the invasive growth and malignant development of human glioma, and may be related to the prognosis and the malignant degree of glioma.

Objective To investigate the different effect of exogenous leptin on estrogen receptor α,β mRNA in human breast tumor tissue in nude mice xenograft models. Methods We made nude mice xenograft models of MCF-7 human breast cancer cells cultured in vitro, then divided them into experimental group of]eptin( n = 30)and control group of normal saline( n = 30)randomly. The models of experimental group were injected subcutaneously the recombinant human leptin for 15 consecutive days, the models of control group were injected subcutaneously the same dose of normal saline. A real- time quantitative RT- PCR assay was developed to quantify the expression of estrogen receptor α, β mRNA in tumor tissue, using the relative quantitative analysis. Results The leptin-intervened nude mice xenograft models were safely established. The relative quantitation of estrogen receptor α mRNA was significantly higher in the leptin group than in the normal saline group ( P ＜ 0.01 ), the relative quantitation of estrogen receptor β mRNA was significantly lower in the leptin group than in the normal saline group ( P ＜ 0. 01 ). Conclusion The nude mice xenograft models can be safely intervened with human leptin by subcutaneous injection around tumor.Estrogen receptor is one of the targets of leptin in the progress of breast cancer. Exogenous human leptin can up- regulate the expression of estrogen receptor α and down- regulate the expression of the estrogen receptor βin nude mice xenograft models of human breast tumor.

Objective To study TMSG-1 and Cyclin D1 expressions in esophageal squamous cell carcinoma (ESCC) and their relevance to the clinicopathological data and prognosis. Methods Immunohistochemistry S-P method was used to examine the expressions of TMSG-1 and Cyclin D1 in pathological specimens of 136 cases of ESCC and 13 cases of normal esophageal mucosa. Contrast study among immunohistochemistry, clinicopathological data and prognosis was also analyzed. Results (1)The positive expression rates of TMSG-1 and Cyclin D1 in ESCC were 52.2% and 65.4%, respectively. The expressions of TMSG-1 and Cyclin D1 in ESCC were significantly higher than that in normal esophageal mucosa (P<0.05). (2)The expressions of TMSG-1 and Cyclin D1 were all related to clinical stage , differentiation degree and lymph node metastasis (P < 0.05). (3)The expression of TMSG-1 was negatively correlated to the expression of Cyclin D1 in ESCC (r=-0.386,P=0.000). (4)The expression levels of TMSG-1 and Cyclin D1 were independent risk factors in patients with ESCC (P < 0.05). Conclusions The abnormal expressions of TMSG-1 and Cyclin D1 may cooperatively play a role in initiation and development of ESCC. Co-examination of TMSG-1 and Cyclin D1 expressions in primary tumors may be useful for predicting prognosis of ESCC.