Objective:To clarify the transitional components in the blood of compound Banlangen Granules; To explore the mechanism of drugs in the treatment of epidemic encephalitis B, hepatitis and parotitis.Methods:The transitional components in blood of compound Banlangen Granules were analyzed by ultra-high performance liquid chromatography-mass spectrometry (UPLC-MS/MS). The regulatory targets and pathways of compound Banlangen Granules in the treatment of epidemic encephalitis B, hepatitis and parotitis were analyzed based on UPLC-MS/MS and network pharmacology.Results:A total of 9 blood components were identified, of which 8 were prototype components, including sucrose, o-aminobenzoic acid, uridine, adenosine, guanosine, indole-3-acetonitrile-2 murine-S-β-D-glucopyranoside and salicylic acid. Through network pharmacological analysis, it was concluded that compound Banlangen Granules may treat epidemic encephalitis B, hepatitis and parotitis by regulating lipid and atherosclerosis, insulin resistance, IL-17 and other signal pathways.Conclusion:The 9 blood components of compound Banlangen Granules may treat epidemic encephalitis B, hepatitis and parotitis by regulating lipid and atherosclerosis, insulin resistance, IL-17 and other signal pathways.

Objective to investigate the role of Pim‐3 and NF‐κB in the development and progression of infiltrating ductal carcinoma of the breast .Methods Here ,we used immunohistochemistry to detect expression of Pim‐3 and NF‐κB in 75 samples of infiltrating ductal breast carcinoma ,21 samples of intraductal breast carcinoma and 30 normal breast tissues .The relationship of their expression ,as well as their correlation with clinicopathological features and patient survival were assessed .Results In con‐trast ,both Pim‐3 and NF‐κB were more commonly detected in infiltrating ductal carcinoma than in intraductal carcinoma and normal tissue .In the infiltrating ductal carcinoma ,the positive expression rate of Pim‐3 was 77 .3% ,and that of NF‐κB was 68 .0% ;in duc‐tal carcinoma of the breast ,the positive expression rate of Pim‐3 was 52 .4% ,and that of NF‐κB was 42 .9% ;in the normal breast tissue ,the positive expression rate of Pim‐3 was 23 .3% ,and that of NF‐κB was 16 .7% ;the positive expression rate of Pim‐3 was correlated with tumor size ,histological grade ,and clinicopathological stage ;and that of NF‐κB was correlated with tumor size ,histo‐logical grade ,lymph node metastasis of breast cancer .Spearman rank correlation analysis revealed a positive correlation between Pim‐3 expression and NF‐κB expression in infiltrating breast cancer (r=0 .243) .Conclusion Our results demonstrate that Pim‐3 and NF‐κB play a role in the initiation and development of breast cancer ,thus ,these proteins may serve as useful diagnostic and prognostic markers of invasive breast cancer .

Objective To observe the influence of losartan on rats′blood pressure and myocardium.Methods To randomly divide 24 4-year-old spontaneously hypertensive male rats into 3 groups.Group A is given normal saline(NS);group B is given losartan 20 mg/kg in 4-week-old;group C is given the same dose in 1 2-week-old;To take another 4-week-old male wistar rats to form group D,feed NS served as normal comparison.Each group was given NS or losartan one time each day through intragastric administration for 4 weeks.Follow up 24 weeks after stopping the medicine.Rats’SBP were measured through tail cuff method respectively when at 4,8,12,16,20 and 24 weeks’ time.After 24 weeks,cut off the head to take the sample,the RVW/BW,HW/BW,LVW/BW were measured through weighing method.To observe the morphologic change of the cardiac muscles and measure the area of myocardium cell cross section through HE stain method.Ra-dioimmunoassay was used to measure the AngⅡand Aldo level of the blood plasma and myocardium.Results At the age of eight-week,the SBP of group A and C are higher than those of group D(P<0.05),group B significantly lower than group A.After stopping the medicine, SBP of group B is slowly increasing and still lower than group A(P<0.05)as well as group C (P<0.05)until the 24th week.At the 16th week,SBP of group C is significantly lower than that of group A(P<0.05)but higher than group D,but quickly increases after stopping the medicine till the 24th week and there is no significant difference between group A and C(P>0.05).About LVW/BW,HW/BW and the ar-ea of myocardium cell cross section,group A increased significantly in comparison with group D (P<0.05),group B and C decreased in comparison with group A(P<0.05),group B decreased significantly in comparison with group C(P<0.05).Through the observation of the area of myocardial cell cross section under the microscopy,group A increased significantly in comparison with group D about myocardial cell fattened and the gap between cells are widened,the fibers of mesenchyma proliferates.group B and D are similar,group C is between group B and A.The level of AngⅡand Aldo in plasma and myocardium of group A increased significantly in comparison with group D (P<0.05),group B decreased significantly in comparison with group A and C(P<0.05).Conclusion To use losartan for a short time in trea-ting rats with prehypertension can get the after-effect of reducing blood pressure and anti-ventricular-remodeling.

Objective To explore the specific cellular and humoral immunity induced by dendritic cells (DC) vaccine loading allogenic microvascular endothelial cell bEnd. 3 antigen against U14 cervical cancer cell of mice. Methods Mouse brain microvascular endothelial cell bEnd. 3 was cultured and identified for preparation endothelial cell bEnd. 3 antigen. The level of mRNA expression of vascular endothelial growth factor receptor 2 (VEGF-R2) and integrin αV was detected by reverse transcription (RT)-PCR. The BALB/c mice were immuned with DC loading bEnd. 3 antigen 4 times in 4 weeks (bEnd. 3-DC group), while the mice only were immuned with DC or injected with phosphate buffer saline (PBS group) as control group. One week after last vaccination, U14 cervical cancer cells were injected subcutaneously into the mice. The tumor size, cytotoxic T lymphocyte (CTL) response of spleen lymphocytes in vitro, the percentage of CD3+ CD+8 surface markers of spleen lymphocytes, and the titer of serum antibody were detected. The specific immunity was examined by immunocytochemistry and western blot. Results The expression of VEGF-R2 and integrin αV gene in bEnd. 3 cells were expressed highly.After the vaccine was injected, the tumors of mice in PBS group grew faster than those in other groups, while the tumors in bEnd. 3-DC group grew slowly and disappeared after 2 weeks. The volume of tumors in DC group grew slower than those in PBS group [(0.11± 0.13) cm3 versus (3.38 ±0.34) cm3]. The CTL response of spleen iymphocytes in vitro showed that bEnd. 3-DC cells could kill bEnd. 3 cells, the special lysis rate was more than 60% . The percentage of CD+3 CD+8 spleen lymphocytes in bEnd. 3-DC group[(38.6 ± 0.7) %] was higher than those in other groups (P ＜ 0.05). The titer of serum antibody of Immunocytochemistry analysis indicated there were specific antigen-antibody reaction to bEnd. 3 cell in bEnd. 3-DC group. Western blot analysis revealed that there were specific bands at 220 000 (VEGF-R2).Conclusions bEnd. 3-DC vaccine can inhibit the tumor growth of U14 cervical cancer cell of mice, which indicates that the special cellular and humorai immunity are induced by bEnd. 3-DC antigen which maybe have some antigens in bEnd. 3 cells that reacts with endothelial cell proliferation-related antigens.

Objective To investigate the differentially expressed genes of primary esophageal squamous cell carci-noma and of normal esophageal mucosa. Methods LCM-GMA-cDNA microarray was used to detect the mRNA from both the primary carcinoma and the corresponding esophageal epithelium in 15 cases of human esophageal squamous cell carcinoma (ESCC). After high-stringent washing, the cDNA microarray was scanned for the fluores-cent signals. Results Among the 886 target genes, 34 genes had significant difference in Ⅰ / Ⅱ and Ⅲ/Ⅳ group. Cell cycle regulators possibly promoting the growth of tumor cells were highly expressed in the early stages of ESCC, whereas adhesion molecules and extracellular matrix-related molecules possibly promoting invasiveness increased in the later stages. Conclusion More than one gene contributed to esophageal cancer. The profiles of gene expression will bring us chance to understanding the molecular mechanism of tumor progression and to support clinical treat-ment.

To screen the genes associated with esophageal cancer, a cDNA microarray technique was established and used for the analysis of the gene expression profile in human esophageal cancer cell line ECa109. The results showed that 107 (12.08%) genes differentially expressed among 886 target genes were identified between ECa109 cell line and normal human esophageal epithelial cells (HEEC), of which 51 (5.76%) were up-regulated and 56 (6.32%) down-regulated. Two genes were validated by quantitative RT-PCR (Q-RT-PCR) and the results were identical. The RNA amplification technique based-T7 RNA polymerase was established. The gene expression profile revealed better consistency between the amplified samples and those without amplification by T7 RNA polymerase, which provides a method for studying the profile of minute quantities of tumor cells in primary esophageal cancers. And the preliminary study on differential expression gene profile also enables us to have an understanding of the pathogenesis and pathomechanism of esophageal cancer.
Carcinoma, Squamous Cell ; genetics ; pathology ; Esophageal Neoplasms ; genetics ; pathology ; Gene Expression Profiling ; methods ; statistics & numerical data ; Gene Expression Regulation, Neoplastic ; Humans ; Oligonucleotide Array Sequence Analysis ; methods ; Reverse Transcriptase Polymerase Chain Reaction ; Tumor Cells, Cultured

Objective To investigate the etfect of microenvironment simulated by colon carcinoma homogenate supernatant on the differentiation and development of human dendritic ceils (DCs), and to investigate the function of vascular endothelial growth factor A (VEGF-A) during this process . Methods Fresh colon carcinoma and peri-cancer tissues were collected to prepare homogenate supernatant. The pe-ripberal blood mononuclear cells were isolated and cultured with 1640 medium including rhGM-CSF and rhIL-4. Then the colon carcinoma homogenate supernatant, peri--carcinoma homogenate supernatant and VEGF-A were added to the cultures at day 2. Antigen of colon carcinoma cell line SW620 was added at day 4 and lipopolysaccharide (LPS) was added at day 6. DCs were collected at day 8 for further study. The con-tent of VEGF-A was tested by ELISA. The morphology and the immunopbenotype of DCs were checked by microscope and flow cytometry, respectively. The expression of CDIa was tested by RT-PCR, and the prolif-eration and killing rate of T cell was measured by CCK-8. Results The content of VEGF-A in the homoge-nate supernatant of colon carcinoma was significantly higher than that of the peri-carcinoma (P ＜ 0. 05). Compared with normal DCs, the cell morphology of colon carcinoma homogenate aupernatant group was in-hibited, and the cell number was decreased. Besides, the positive expression rate of DC surface markers de-creased (P ＜ 0.01). The capacity of mixed lymphocyte reaction (MLR) and killing capacity of T cells de-creased(P ＜0.01). However, there was almost no difference between VEGF-A group and normal DCs on the cell morphology and cell number, and VEGF-A had no obvious inhibition on the expression of DCs sur-face markers (P ＞ 0.05). But VEGF-A group had significantly inhibitory effect on the MLR and T cells kill-ing. Conclusion The tumor microenvironment simulated by the colon carcinoma homogenate supernatant obviously has inhibitory effect on the differentiation and function of DCs, and VEGF-A has the inhibitory effect on DC function, but the inhibitory effect is not through the inhibition of the expression of DC costimu-lators.

The differentially expressed genes between esophageal squamous cell carcinoma (ESCC) with or without lymphatic metastasis were investigated by gene chip, and the lymphatic metastasis-associated genes were screened out. Expression array was used to detect the mRNA from both the primary carcinoma and the corresponding esophageal epithelium in 15 cases of human ESCC. The lymphatic metastasis-associated genes were screened by bioinformatics between ESCC with or without lymphatic metastasis. The results showed that 43 (4.85 %) genes significantly differed between the ESCC with and without lymphatic metastasis (P<0.05), of which 18 (2.03 %)were upregulated and 25 (2.82 %) down-regulated. The up-regulated genes were involved in cell adhesion molecules and cell membrane receptors and the down-regulated genes were mostly cell cycle regulators and intracellular signaling molecules. It was suggested that lymphatic metastasis-associated genes were screened by gene chip, which was helpful to understand the molecular mechanism of ESCC lymphatic metastasis and lymphatic metastasis-associated genes might be used as diagnostic markers and therapeutic targets for lymphatic metastasis.

The differentially expressed genes between esophageal squamous cell carcinoma (ESCC)with or without lymphatic metastasis were investigated by gene chip, and the lymphatic metastasisassociated genes were screened out. Expression array was used to detect the mRNA from both the primary carcinoma and the corresponding esophageal epithelium in 15 cases of human ESCC. The lymphatic metastasis-associated genes were screened by bioinformatics between ESCC with or without lymphatic metastasis. The results showed that 43 (4.85%) genes significantly differed between the ESCC with and without lymphatic metastasis (P＜0.05), of which 18(2.03%)were upregulated and 25 (2.82 %) down-regulated. The up-regulated genes were involved in cell adhesion molecules and cell membrane receptors and the down-regulated genes were mostly cell cycle regulators and intracellular signaling molecules. It was suggested that lymphatic metastasis-associated genes were screened by gene chip, which was helpful to understand the molecular mechanism of ESCC lymphatic metastasis and lymphatic metastasis-associated genes might be used as diagnostic markers and therapeutic targets for lymphatic metastasis.

Objective To evaluate the choice of therapeutic methods and MRI diagnosis of acoustic neuroma.Methods The MRIimaging and clinical materials of 60 patients with acoustic neuroma were analyzed,48 cases were underwent an operation,12 cases weretreated conservatively or gamma knife treatment,follow-up ranged from 1 to 4 years.Results There were 62 tumors round the internalauditory canal.There were 58 cases with single acoustic neuroma and two cases with couples acoustic neuroma.28 tumors demonstratedhypointense and 30 tumors demonstrated hypo-and isointense signals on T_1 weighted image.38 tumors demonstrated hyperintense and 24tumors demonstrated hyper-and isointense signals on T_2 weighted image.The Ⅶ,Ⅷ nerves affected side were thickened than that ofopposite side in 32 patients.After Gd-DTPA administration 24 tumors were homogeneously enhanced,26 tumors were inhomogeneously or circularly enhanced in 50 acoustic neuromas of 48 cases.Operation was still the main election for acoustic neuroma.Conclusion MRI is an effective method in the diagnosis of acoustic neuroma,and providing advice for clinics in making therapeutic programs.