Establishment of zebrafish osteopenia model induced by dexamethasone / 药学学报

Zebrafish was selected as model animal, and glucocorticoid dexamethasone was used as a model compound to establish a rapid and high efficient osteopenia model. Zebrafish larvae at 4 days post fertilization (dpf) were exposed to a serial concentrations of dexamethasone solutions, and 0.5% DMSO was selected as the vehicle control group. All groups were incubated in 24-well plates (28.5 degrees C) until 9 dpf. In addition, effects of 10 micromol x L(-1) dexamethasone on preventing against osteopenia induced by etidronate disodium were also investigated. Zebrafish bones at 9 dpf were stained with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of bone mineralization. Results showed that dexamethasone group at 2.5, 10 and 25 micromol x L(-1) can decrease the staining area and the staining optical density values of zebrafish head bones when compared with the vehicle control group (0.5% DMSO), which suggested that dexamethasone can significantly reduce the zebrafish mineralized bone and the bone mineral density. Results also showed that 15 and 30 microg x mL(-1) etidronate disodium can increase the mineralized matrix of zebrafish head bone and prevent against osteopenia induced by dexamethasone. In conclusion, the study indicated that zebrafish can be an idea osteopenia model induced by dexamethasone.

Metabolism study of asperosaponin VI by using zebrafish / 药学学报

Model organism zebrafish was used to study metabolism of asperosaponin VI from Dipsacus asper Wall. ex Henry for the first time. Metabolic components of asperosaponin VI after exposing to zebrafish for 24 h were identified by high performance liquid chromatography--electrospray mass spectrometry (HPLC-ESI-MS), the separation was performed with a Zorbax C18 column using a binary gradient elution of 0.05% formic acetonitrile--0.05% formic acid water. The quasi-molecular ions of compounds in both negative and positive mode were observed for molecule mass information, and the potential structures were identified by attentive study on the deglycosylation metabolites and one hydroxylation metabolite of asperosaponin VI. The results were highly in consistent with metabolism of asperosaponin VI in rat. It can be concluded that zebrafish model can wonderfully imitate current metabolic model with advantages of small amount of lower cost, far less amount compound, higher efficiency and more simple, and can reflect integrated metabolism results of in vivo method. Zebrafish metabolic model may become a novel organism model for quick predication on metabolism of even mircoamount compound, which can enrich the available models greatly.

Lymphatic targeting study of pingyangmycin-activated carbon nanoparticles treating oral cancer lymph node metastasis / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the drug distribution in tissues of cervical lymph node metastasis mice model after submucosa adjacent cancer injection of pingyangmycin-activated carbon nanoparticles (PYM-CH-NP) and evaluate the lymph targeting effect of PYM-CH-NP.</p><p><b>METHODS</b>Pingyangmycin (PYM) was radiolabeled with 125I by modified the chloramine T method. Cervical lymph node metastasis mice model was established by buccal submucosa inoculation of a high lymph metastasis cell line U14 cancer cell. 360 mice models burdened with cervical lymph metastasis were randomly divided into 3 groups. PYM group was treated with PYM water solution, PYM-CH-NP group was treated with PYM-CH-NP. Negative control group was injected with activated carbon nanoparticles. PYM-CH-NP and pingyangmycin water solution were injected in pericancer submucosa of the mice respectively. The radioactivity of drug in blood, heart, liver, spleen, lung, kidney and cervical lymph node were measured after 0.5, 1, 4, 8, 12, 24, 48, 72, 96, 120, 144, 168 h administration. The radioactivity of each samples per unit weight were calculated. The selectivity index (SI) and targeting index (TI) of drug were calculated.</p><p><b>RESULTS</b>The radioactivity of drug in cervical lymph node of PYM-CH-NP group was much higher than PYM group in each time point (P < 0.001), whereas the blood, heart, liver, spleen, lung and kidney uptake of pingyangmycin was greatly decreased in PYM-CH-NP group after 4 h administration (P < 0.001). The SI value of PYM group at each time point was less than 1. While the minimum SI and TI value of PYM-CH-NP was 1.793 and 1.562, the maximum value reached to 68.126 and 14.623 after 72 h administration.</p><p><b>CONCLUSION</b>PYM-CH-NP can increase drug dosage in metastasized cervical lymph nodes, and decrease drug dosage of other organs. So better therapeutic outcome and little adverse reaction may be achieved for lymph node metastasis.</p>

Anticancer effects of Pingyangmycin-activated carbon nanoparticles against human oral squamous carcinoma Tca8113 and BcaCD885 cell lines in vitro / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The cytotoxic effects of a new formulation of Pingyangmycin-activated carbon nanoparticles (PYM-CH-NP) on two human oral squamous cell carcinoma Tca8113 and BcaCD885 cell lines were studied in vitro.</p><p><b>METHODS</b>The inhibitory effects of PYM-CH-NP and Pingyangmycin (PYM) were evaluated by methyl thiazolyl tetrazolium (MTT) assay at 1-7 days. The 50% inhibition concentration values (IC50) and relative antitumor activity (RAA) of PYM-CH-NP and PYM against Tca8113 and BcaCD885 with different drug concentration were evaluated. The time-dependent cytotoxic effects of PYM-CH-NP and PYM were during 1-5 days, so the doseeffect relationship was investigated at 5th day.</p><p><b>RESULTS</b>Both PYM-CH-NP and PYM had high anticancer effects on Tca8113 and BcaCD885, and the cytotoxic effects were dose-dependent and time-dependent.</p><p><b>CONCLUSION</b>The activated carbon nanoparticles (CH-NP) may serve as a new drug delivery carrier of PYM. The new formulation PYM-CH-NP could slow down drug release, prolonged the drug concentration and its acting time, so more effective anticancer efficacy could be achieved.</p>

Correlation between the expression of thrombospondin-1 and neovascularization in mucoepidermoid carcinoma / 中华医学杂志(英文版)

<p><b>BACKGROUND</b>Researchers have recently demonstrated that thrombospondin-1 (TSP-1) has an important function in regulating neovascularization. Whether it inhibits or accelerates neovascularization, however, is still controversial. We found few reports about the correlation between TSP-1 and vascularization in mucoepidermoid carcinoma. In this research, the distribution and expression of TSP-1 in mucoepidermoid carcinoma were investigated. We also analyzed (1) the correlation between the expression of TSP-1 and microvessel density (MVD), as an indicator of neovascularization activity, and (2) the effect of TSP-1 on neovascularization and tumor growth in the subcutaneous xenotransplanted model of mucoepidermoid carcinoma.</p><p><b>METHOD</b>(1) The sites and intensity of expression of TSP-1 and the MVD were analyzed in 45 cases of mucoepidermoid carcinoma after surgery by the method of streptavidin-peroxidase (SP) immunohistochemistry; and (2) recombinant human thrombospondin-1 (rhTSP-1) was injected twice a week for five consecutive weeks around the tumor in the subcutaneous xenotransplanted tumor model of mucoepidermoid carcinoma in nude mice. Each week, the tumor size was measured, in order to draw the growth curve of the xenotransplanted tumor model of mucoepidermoid carcinoma, and MVD was measured.</p><p><b>RESULTS</b>(1) The positive expression of TSP-1 protein was 57.78% (26/45). Most positive staining for TSP-1 was found in the cytoplasm of the cancer cells, while some staining occurred in the extracellular matrix. The mean MVD in 45 cases of mucoepidermoid carcinoma was 58.17 +/- 19.77 per 100 visual fields. Tumors with a high expression of TSP-1 showed a low MVD value, and the TSP-1 immunocompetence and microvessel density showed a significant negative correlation (r(s) = -0.947, P < 0.001). (2) The xenotransplanted tumors with the injection doses of 1.25, 0.75 and 0.25 microg/ml respectively were 36.97%, 53.36% and 73.61% of the size of the control group ((451 +/- 92), (651 +/- 113), (898 +/- 86) and (1220 +/- 157) mm(3) respectively, F = 53.167, P < 0.001), and their weights were respectively 35.14%, 51.35% and 70.27% of the control group ((1.3 +/- 0.5), (1.9 +/- 0.5), (2.6 +/- 0.3), and (3.7 +/- 0.7) g respectively, F = 62.669, P < 0.001). Their MVDs were 25.00%, 45.93%, and 72.20% respectively of the control group and concentration dependent (15.43 +/- 3.45, 28.35 +/- 4.24, 44.57 +/- 3.35 and 61.73 +/- 5.43 per 100 visual fields respectively, F = 54.582, P < 0.001).</p><p><b>CONCLUSIONS</b>The TSP-1 has a higher expression in mucoepidermoid carcinoma and the expression has a significant negative correlation with neovascularization. The TSP-1 inhibits neovascularization and tumor growth, and it might be a new biological therapy for treatment of patients with mucoepidermoid carcinoma.</p>

The expression of inhibitor-1 of DNA binding/differentiation-1 and thrombospondin-1 in mucoepidermoid carcinoma of different malignant degree / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the expression of inhibitor-1 of DNA binding/differentiation-1 (Id-1) and thrombospondin-1 (TSP-1) genes in mucoepidermoid carcinoma of different malignant degree and analyze the relationship between them.</p><p><b>METHODS</b>Using immunohistochemistry (IHC) staining technique, TSP-1 and Id-1 proteins in the mucoepidermoid carcinoma of different malignant degree, including well-differentiated, moderately differentiated and poorly differentiated mucoepidermoid carcinoma, and normal salivary gland tissues were detected.</p><p><b>RESULTS</b>The positive rate of Id-1 and TSP-1 in normal salivary glands were apparently lower than that in malignant mucoepidermoid carcinoma(P = 0.000, P = 0.013). The positive rate of Id-1 in moderately and poorly differentiated mucoepidermoid carcinoma was higher than that of the well-differentiated (P = 0.001, P = 0.002). However, the positive expression of Id-1 showed no relationship between the moderately and poorly differentiated mucoepidermoid carcinoma(P > 0.05). The positive rate of TSP-1 in poorly differentiated mucoepidermoid carcinoma was less than that of the well-differentiated(P = 0.014). The positive expression of TSP-1 showed no relationship between the moderately and poorly differentiated mucoepidermoid carcinoma(P > 0.05), and the positive expression of it also showed no relationship between the moderately and well differentiated mucoepidermoid carcinoma (P > 0.05). The expression of Id-1 and TSP-1 showed negative correlation(r = -0.394, P = 0.002).</p><p><b>CONCLUSION</b>The expression of TSP-1 may inhibit the development of the mucoepidermoid carcinoma, contrarily, the expression of Id-1 may prompt the development of the mucoepidermoid carcinoma. The expression of Id-1 and TSP-1 has negative correlation.</p>

Study of target pegylated recombinant mutant human granulocyte colony stimulating factor / 生物工程学报

Recombinant mutant human granulocyte colony stimulating factor (rmhG-CSF) was pegylated, purified and characterized. rhG-CSF was mutated in position 1,3,4,5,17, and cysteine was added in C-terminal. rmhG-CSF was pegylated by PEG-Mal 20000 and separated by ion-exchange chromatography, gel filtration chromatography. Analysis of SDS-PAGE showed thar the purity of the separated PEG-rmhG-CSF was greater than 95%. and in intro and in vivo bioactivity study showed that target modified PEG-rmhG-CSF kept full bioactivity which was better than traditional pegylation method, and longer half-life was proved in mice.

A retrospective clinicopathologic study of 30 cases of sublingual gland malignant tumors / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To assess the clinical and histological features and therapeutic efficacy of 30 cases of malignant sublingual salivary gland tumors.</p><p><b>METHODS</b>The clinicopathologic data of 30 patients with malignant sublingual salivary gland tumor were obtained from West China Hospital of Stomatology, Sichuan University from 1955 to 2005.</p><p><b>RESULTS</b>There were 18 male and 12 female, and the average age of patients was 50.6 years old. Seventeen cases were adenoid cystic carcinoma, accounting for 56.7%. There were 17 cases clinically staged as III, accounting for 56.7%. Distant metastasis and tumor recurrence were the main death reasons. The overall local recurrence rate was 30.0%, and distant metastasis rate was 26.7%.</p><p><b>CONCLUSION</b>Sublingual gland malignant tumors are rare and most of them are adenoid cystic carcinoma. Surgery is the main treatment option. The resection of the tumor accompanying with the neck dissection is the key method to achieve good therapeutic effect. The postoperative radiotherapy and chemotherapy should be adjuvant.</p>

An experirmental study on the preparation and drug sustained release characteristics of Pingyangmycin Albumin Microspheres / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The aim of this study was to prepare Pingyangmycin Albumin Microspheres (PYM-AMS) for arteriovenous malformations treatment.</p><p><b>METHODS</b>PYM-AMS was prepared at 140 degrees C by the method of emulsification-heat solidification and its characteristics were evaluated, such as morphosis, particle size, drug loading (DL%), encapsulation efficiency (EE%), stability and drug sustained-releasing in vitro. After being packaged, PYM-AMS were sterilized with 13.7 kGy of 60Co. Small samples of PYM-AMS were packaged in small bottles and stored at 3 - 5 degrees C, 15 - 25 degrees C, 37 degrees C for 3 months, then checked the change of morphology, DL, EE and the release rate.</p><p><b>RESULTS</b>The surface of particles was smooth and integrated. The average diameter of PYM-AMS particles was 139.422 microm and 80% was in the range of 56 - 251 microm. The mean DL% and EE% were 26.47% and 84.3%, respectively. PYM released fast in 5 h, but then released slowly. 88.65% drugs were released in 24 h, and t50 was 1.5 h. There was no obvious change of the morphology, DL,EE and the release rate of PYM-AMS stored at 3 - 5 degrees C 15 - 25 degrees C, 37 degrees C for 3 months.</p><p><b>CONCLUSION</b>PYM-AMS prepared in this study had sustained-release effect, high drug loading and high stability. Albumin is a good carrier of PYM embolization agent.</p>

The changes of bacteria group on oral mucosa after radiotherapy of postoperative patients of oral carcinoma / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the microbial contents presented on the surface of mucosa in the oral cavity of patients who accepted radiotherapy, and to provide the evidences of controlling post-radiotherapeutic infections.</p><p><b>METHODS</b>32 patients (19 males and 13 females) aged from 37 - 72 received radiotherapy after oral squamous cell carcinomas operation were selected. Samples of saliva were obtained from the radiated center and opposite mucosa before and after radiotherapy. The detective amount, detective ratio and constituent ratio were analysed by cultivation and identification.</p><p><b>RESULTS</b>Streptococci, Candida albicans and Pseudomonas aeruginosa significantly increased on both sides of the oral mucosa while Neisseria and Actinobacillus decreased on radiated region after the radiotherapy.</p><p><b>CONCLUSION</b>Radiotherapy has great effects on oral bacteria and pathogenic organism may play a role in post-radiotherapy infections. It is necessary to do bacteria culture and choose sensitive antibiotics regularly for post-radiotherapeutic patients.</p>

An experimental study on PLGA carrier for oral tissue engineered mucosa by subcutaneous implantation in rabbits / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To look for the best carrier for cultivating oral tissue engineered mucosa.</p><p><b>METHODS</b>A series of membrane of scaffold materials of polycleclide-co-plycoclide (PLGA) were studied on their weight and biocompatibility after they had been implanted subcutaneously in rabbits for 1, 2, 3, 4 weeks respectively.</p><p><b>RESULTS</b>PLGA I , II, III degraded completely in rabbits after 2, 3, 4 weeks respectively. The other PLGA membrane degraded about 50% after 4 weeks. Histologically, the reactions of PLGA I, II, III with surrounding tissues were normal and membranes had a good biocompatibility.</p><p><b>CONCLUSION</b>The biodegrading rate of PLGA II is suitable for clinic practice. PLGA II was a promising carrier for oral tissue-engineered mucosa due to its excellent biocompatibility and biodegrading rate.</p>

Studies on the transfection of umbilical endothelia with catalytic subunit of telomerase / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the variety of proliferating ability of umbilical endothelia (UE) transfected by plasmid pBABE-HYGR-hTERT.</p><p><b>METHODS</b>UE was identified from two aspects: morphology and CD34 labeling technique. The plasmid was obtained and identified by alkali splitting and gel electrophoresis. Liposomes were used to transfect UE. RT-PCR based telomeric repeat amplification protocol (TRAP) assay was used to measure the telomerase activity of endothelia.</p><p><b>RESULTS</b>UE arranged as "cobblestone" and were positive of CD34 labeling. Endothelia transfected by pBABE-HYGR-hTERT(HC) had an raised absorbance of 0.889. The shape of growth curve of HC was similar to UE. But the absorbance of MTT test and the amount of HC were prior to UE at every measuring time and the amount of HC increased four times within 8 days (P < 0.05).</p><p><b>CONCLUSION</b>The transfection of pBABE-HYGRO-hTERT had greatly improved the proliferating abilities and activated the telomerase of UE.</p>

Expression of nucleolar organizer regions associated proteins of T lymphocytes in the patients with oral and maxillofacial tumors / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To study the Ag-NORs expressive character of T lymphocyte in patients with oral and maxillofacial tumor and evaluate its diagnostic value for the patients.</p><p><b>METHODS</b>Nucleolar organizer regions(NORs) of T lymphocyte from 86 normal adults, 102 patients with oral and maxillofacial benign tumors and 87 patients with oral and maxillofacial malignant tumors were analyzed through KL imaging system.</p><p><b>RESULTS</b>There was significant difference among normal adults, benign and malignant patients (P < 0.01). Their average I.S% values were 7.87 +/- 0.12, 5.72 +/- 0.14, 4.19 +/- 0.13, respectively.</p><p><b>CONCLUSION</b>The expression of Ag-NORs of T lymphocyte has definite relation with oral and maxillofacial tumors; detection of Ag-NORs might play valuable in diagnosis of oral and maxillofacial tumors.</p>

Preparation of lymphatic targeting dosage form: pingyangmycin absorbed on activated carbon nanoparticles / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To prepare anticancer nanoparticles for targeting therapy for oral cancer lymph node metastasis.</p><p><b>METHODS</b>The activated carbon nanoparticles (CH-NP) were prepared for drug carrier. Pingyangmycin (PYM), a high sensitive anticancer drug for oral squamous cell carcinoma, were selected as model drug. The activated carbon nanoparticles and PYM were mixed with saline and shaken 20 minutes so that PYM was absorbed on activated carbon enough, resulting in a new formulation of PYM (PYM-CH-NP). The absorbency of PYM on activated carbon nanoparticles was evaluated.</p><p><b>RESULTS</b>The diameter distribution for CH-NP ranged form 136 nm, to 540 nm, the average diameter was 176 nm. The proportion of CH-NP to PYM was increased and more absorbency of PYM on activated carbon nanoparticles was achieved.</p><p><b>CONCLUSION</b>The activated carbon nanoparticles has high absorbency of PYM. The new formulation PYM-CH-NP can be used as targeting therapy of cervical lymph node metastasis by peri-cancer submucosal injection.</p>

Study of the relative factors of the biological behavior of pleomorphic adenoma in parotid / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To study the growth way of parotid pleomorphic adenoma and the relative factors.</p><p><b>METHODS</b>The histological slides of 97 cases of the primary parotid pleomorphic adenoma were examined for the state of intra-capsule infiltration and extra-infiltration. The relative relationships between the infiltration state and the histological type, relative amount of various components, size and course of the tumor were analysed to investigate the growth way and relative factors.</p><p><b>RESULTS</b>1. There were more chances to develop infiltration of tumor in which the major content was epithelium. The tumor was severer with the increasing of epithelium, and decreasing of mucous content and elongation of course of the disease. 2. The limitation of the extra-envelop infiltration and budding was 0.085 - 0.210 mm, so, the boundary of partial parotidectomy should be away from the 1 cm envelop.</p><p><b>CONCLUSION</b>The growth way of parotid pleomorphic adenoma is related to the histological types and characters, relative amount of various components and the course of the tumor.</p>

Transfection of the nm23-H1 gene into BcaCD885 cell line inhibits the potential of invasion, adhesion and mobility / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To transfect nm23-H1 into the BcaCD885 cell lines in order to get safe high-efficiency and low-toxicity, and to find out whether nm23-H1 could affect the invasion and metastases ability of BcaCD885 cell lines.</p><p><b>METHODS</b>Lipofect was used to transfect nm23-H1 into BcaCD885 cell lines; immunohistochemistry was used to detect the difference expression of nm23-H1 between transfected and non-transfected cell lines; then transwell-room and wash way were used to detect the difference of invasion and metastases ability between transfected and non-transfected cell lines.</p><p><b>RESULTS</b>PCMV-NEO-BAM system gave the stability expression of nm23-H1; there was significant different NDPKA expression between transfected and non-transfected BcaCD885 cell lines; the invasion and metastases ability of transfected BcaCD885 cell lines decreased obviously.</p><p><b>CONCLUSION</b>nm23-H1 can inhibit the metastases of BcaCD885 cell lines significantly.</p>