[Screening on antiosteoporotic active parts of dipsacus radix based on zebrafish model].

OBJECTIVE: Prednisolone-induced osteoporosis model using zebrafish was used to screen the antiosteoporotic active parts of Dipsacus Radix, in order to investigate the applicability and rationality of the zebrafish model of osteoporosis. METHODS: Zebrafish larvae at 5 days post fertilization (dpf) were exposed with 25 micromol/L prednisolone and 0.5% DMSO for 48 h, then except one group of 25 micromol/L prednisolone, other groups of 25 micromol/L prednisolone were treated with a range of concentration (0.025, 0.25, 2.5, 25 microg crude drug/mL) of extract of Dipsacus Radix and its different concentration ethanol elution parts of macroporous resin with 25 micromol/L prednisolone. All groups were incubated in 24-well plates (28.5 degrees C) until 10 dpf. Zebrafish skeleton at 10 dpf were anesthetized and fixed for staining with alizarin red. Quantitative analysis of the stained area was performed by microscopic inspection and digital imaging methods to reflect the amount of zebrafish head skeleton mineralization. RESULTS: The results indicated that head skeleton mineral area and integrated optical density (IOD) of 25 micromol/L prednisolone model group were significantly decreased when compared with vehicle control group, and the extract of Dipsacus Radix and its 30%, 50%, 70% and 90% ethanol elution parts of macroporous resin rescued the further bone loss of zebrafish induced by prednisolone when compared with the model group. HPLC analysis indicated that components of 30%, 50%, 70% and 90% ethanol elution parts of macroporous resin containing saponins and nonsaponins components. CONCLUSION: Both saponins and nonsaponins can prevent bone loss of zebrafish induced by prednisolone. This novel osteoporosis zebrafish model was successfully used to screen antiosteoporotic active parts of Dipsacus Radix, which had advantages of simple, high efficiency and easy to perform.

[Establishment of zebrafish osteopenia model induced by dexamethasone].

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].

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].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

Derived vascular endothelial cells induced by mucoepidermoid carcinoma cells: 3-dimensional collagen matrix model.

Mucoepidermoid carcinoma undergoes uniquely vigorous angiogenic and neovascularization processes, possibly due to proliferation of vascular endothelial cells (ECs) induced by mucoepidermoid carcinoma cells (MCCs) in their three-dimensional (3D) microenvironment. To date, no studies have dealt with tumor cells and vascular ECs from the same origin of mucoepidermoid carcinoma using the in vitro 3D microenvironment model. In this context, the current research aims to observe neovascularization with mucoepidermoid carcinoma microvascular ECs (MCMECs) conditioned by the microenvironment in the 3D collagen matrix model. We observed the growth of MCMECs purified by immunomagnetic beads and induced by MCCs, and characteristics of tubule-like structures (TLSs) formed by induced MCMECs or non-induced MCMECs. The assessment parameters involved the growth curve, the length, the outer and inner diameters, and the wall thickness of the TLSs, and the cell cycle. Results showed that MCCs induced formation of the TLSs in the 3D collagen matrix model. A statistically significant difference was noted regarding the count of TLSs between the control group and the induction group on the 4th day of culture (t=5.00, P=0.001). The outer and inner diameters (t(1)=5.549, P(1)=0.000; t(2)=10.663, P(2)=0.000) and lengths (t=18.035, P=0.000) of the TLSs in the induction group were statistically significant larger than those in the control group. The TLSs were formed at the earlier time in the induction group compared with the control group. It is concluded that MCCs promote growth and migration of MCMECs, and formation of the TLSs. The 3D collagen matrix model with MCMECs induced by MCCs in the current research may be a favorable choice for research on pro-angiogenic factors in progression of mucoepidermoid carcinoma.

[Anticancer effects of Pingyangmycin-activated carbon nanoparticles against human oral squamous carcinoma Tca8113 and BcaCD885 cell lines in vitro].

OBJECTIVE: 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. METHODS: 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. RESULTS: Both PYM-CH-NP and PYM had high anticancer effects on Tca8113 and BcaCD885, and the cytotoxic effects were dose-dependent and time-dependent. CONCLUSION: 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.

[Intralesional Pingyangmycin therapy for 51 infantile patients with parotid gland hemangiomas].

PURPOSE: To investigate the indication and outcome of intralesional Pingyangmycin (PYM) therapy for parotid gland hemangiomas in early childhood. METHODS: 51 infantile patients with hemangiomas in the parotid gland were studied retrospectively, which had been treated in the clinic of West China Hospital of Stomatology, Sichuan University during the 15-year period from May 1990 to May 2005. In this study, 21 were male, 30 were female, and the ratio of male to female was 1:1.43. The age of the children ranged from 6 months to 4 years, with an average age of 10 months. 38 were deep-seated hemangiomas, and 13 were mixed hemangiomas. 27 were in the right parotid gland and 24 in the left, no bilateral case. All the patients underwent intralesional injection of a solution of 8 mg PYM in 8 mL normal saline mixed with 5 mg dexamethasone. The total dose of PYM ranged from 20mg to 35 mg, which was administered 0.5 or 1 mg per injection. SPSS10.0 software package was used to compare the treatment efficacy between the patients with hemangioma <4 cm in diameter and >or=4 cm in diameter. RESULTS: Hemangiomas of 42 cases (82.35%) showed complete resolution with good appearance, 8 cases (15.69%) were partly regressed, and 1 case (1.96%) had no obvious size change. No serious side effects were seen, such as pulmonary fibrosis and growth inhibition. No significant correlation was found between treatment efficacy and tumor size. CONCLUSION: Intralesional PYM therapy maybe is a selective primary therapy option for parotid gland hemangioma, and ultrasonography should be useful for diagnosis and treatment of this lesion.

The expression profile of microRNAs in a model of 7,12-dimethyl-benz[a]anthrance-induced oral carcinogenesis in Syrian hamster.

BACKGROUND: Non-coding RNA molecules, such as microRNAs, may play an important role in carcinogenesis. Recent studies have indicated that microRNAs are involved in initiation and progression of various malignancies. However, little work has been done to compare the microRNA expression patterns in oral cancer. In this study, we constructed an animal model of oral squamous cell carcinoma to investigate expression profiles of microRNAs in oral carcinogenesis. METHODS: The animal model of oral squamous cell carcinoma was conducted by tri-weekly (Monday, Wednesday, and Friday) painting with 5% DMBA in acetone. Six Syrian hamsters, including three from the treated group and three from the control group, were used as a training group for microRNA microarray analysis. All microarray data were analyzed by Significance Analysis of Microarrays (SAM) and CLUSTER 3.0 software, and this result was further confirmed by qRT-PCR assay. RESULTS: Seventeen microRNAs were differentially expressed in oral squamous cell carcinoma. Five microRNAs (hsa-miR-21, hsa-miR-200b, hsa-miR-221, hsa-miR-338, and mmu-miR-762) were significantly upregulated and twelve microRNAs (hsa-miR-16, hsa-miR-26a, hsa-miR-29a, hsa-miR-124a, hsa-miR-125b, mmu-miR-126-5p, hsa-miR-143, hsa-miR-145, hsa-miR-148b, hsa-miR-155, hsa-miR-199a, and hsa-miR-203) were down-regulated in cancer tissues. The expression levels of hsa-miR-21 and hsa-miR-16 seen with Stem-loop qRT-PCR were also seen in microarray analysis in all samples. CONCLUSION: Our findings identified specific microRNA expression in oral squamous cell carcinoma and suggested that microRNAs have a role in oral carcinogenesis.

[Embolization and degradation of arteries in vivo by pingyangmycin albumin microspheres].

OBJECTIVE: To investigate the embolization effect of Pingyangmycin-albumin microspheres (PYM-AMS) on small arteries and its process of degradation in vivo. METHODS: Twenty four Japanese white rabbits were randomly divided into four groups, 6 in each group. PYM hydrochloride + 0.9% NaCl, PYM + soybean oil, and PYM-AMS + soybean oil were injected into the central auricular arteries of the rabbits in the three experimental groups, respectively, for about 30 seconds (0.26 mL/per ear, which contained PYM 5 mg/mL). The vessel samples were taken and examined at 2, 7, 14, and 21 days. RESULTS: The PYM + 0.9% NaCl group had no significant vessel changes. In the PYM+ soybean oil group, some endothelial cells dropped off at the 7th day after injection. At the 21st day, mild proliferation of endothelial cells and walls of central auricular arteries were observed, especially on the intima. But the lumen was still obvious and the blood flow was not blocked. In the PYM-AMS group, the central auricular arteries were narrowed at the 7th day after injection. At the 21st day, the vessels had sclerostenosis, and the blood flow was blocked. At the 14th day, significant proliferation of endothelial cells and walls of central auricular arteries were observed. The surface of PYM-AMS was absorbed. At the 21st day, the walls of central auricular arteries and some small veins proliferated obviously, and the arteries were sclerostenosed. Many smooth muscle cells, fibroblasts, and myofibroblasts in the original blood vessel lumen appeared. There were thrombi besides the PYM-AMS. CONCLUSION: PYM-AMS may become an option for the treatment of large venous or arteriovenous malformations and for the local chemotherapy of malignant tumors.

Correlation between the expression of thrombospondin-1 and neovascularization in mucoepidermoid carcinoma.

BACKGROUND: 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. METHOD: (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. RESULTS: (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). CONCLUSIONS: 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.

[Correlation between the expression of thrombospondin-1 and the angiogenesis in mucoepidermoid carcinoma].

OBJECTIVE: To study the relationship between the expression of thrombospondin-1 (TSP-1) and the angiogenesis in mucoepidermoid carcinoma. METHODS: Immunohistochemistry were applied to detect the expression of TSP-1 and the value of microvessel density (MVD) in 45 mucoepidermoid carcinoma patients. RESULTS: Positive expressions of TSP-1 protein were detected in 26 of the 45 (57. 78%) cases. Most positive staining for TSP-1 was observed in the cytoplasm of the cancer cells, some of those were in the extracellular matrix. The mean MVD in 45 cases with mucoepidermoid carcinoma was 60. 68 +/- 19.84 vessels per 100 field of vision. Tumors with a high expression of TSP-1 showed a low value of MVD and the correlation between TSP-1 immunocompetence and microvessel density was highly significant (r(s) = -0.942, P < 0.001). CONCLUSION: The TSP-1 is expressed in most mucoepidermoid carcinoma and were associated with neovascularization. TSP-1 is likely to inhibit the extensive neovascularization and increased TSP-1 expression might inhibit angiogenic phenotype in mucoepidermoid carcinoma.

[The expression of inhibitor-1 of DNA binding/differentiation-1 and thrombospondin-1 in mucoepidermoid carcinoma of different malignant degree].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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). CONCLUSION: 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.

[Study of target pegylated recombinant mutant human granulocyte colony stimulating factor].

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].

OBJECTIVE: To assess the clinical and histological features and therapeutic efficacy of 30 cases of malignant sublingual salivary gland tumors. METHODS: 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. RESULTS: 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%. CONCLUSION: 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.

[The changes of bacteria group on oral mucosa after radiotherapy of postoperative patients of oral carcinoma].

OBJECTIVE: 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. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[An experirmental study on the preparation and drug sustained release characteristics of Pingyangmycin Albumin Microspheres].

OBJECTIVE: The aim of this study was to prepare Pingyangmycin Albumin Microspheres (PYM-AMS) for arteriovenous malformations treatment. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Effects of Pingyangmycin albumin microspheres on sclerostenosis of the rabbit central auricular arteries].

PURPOSE: The aim of this study was to investigate the mechanism of Pingyangmycin-albumin microspheres (PYM-AMS) acted on the rabbit central auricular arteries. METHODS: In the study of randomized block design, 24 Japanese white rabbits were divided into 4 groups, 6 rabbits in each group. The animals were put on the operation table after being anaesthetized by intramuscular injection of Xumianxin (0.2 ml/Kg) and the interior division was blocked to stop drug liquid get into the interior ear. After being sterilized with 75% alcohol, No.1(PYM hydrochloride injection+0.9% NaCl), No.2(PYM+soybean oil), No.3(PYM-AMS+soybean oil) liquids which contained PYM 5mg/ml were injected into the central auricular arteries of the animals about 30 seconds (0.26 ml/per ear), respectively. Then these vessels were examined histologically after 2,7, 14, 21 days respectively. RESULTS: After injection, in PYM+0.9% NS control group (No.1 liquid), the ears and vessels had no significant changes. In PYM+soybean oil control group (No.2 liquid), at the 2nd day, the endothelial cells were mild swollen. At the 7th day, some endothelial cells were dropped off. At the 14th day, the central auricular arteries had mild change, but the blood flow was not blocked. At the 21st day, the wall of the central auricular artery had more layers, especially on the intima, but the lumen was still obvious. A few of endothelial cells were proliferative. In PYM-AMS group(No.3 liquid), the blood was not resumed at once, and there was some oil in the vessels. At the 1st day, the injection site was mild swollen, and small thrombosis was observed in small vessels. At the 7th-14th day, the central auricular arteries were narrowed, but the blood could be observed. At the 21st day, the vessels had sclerostenosis, and the blood flow was blocked, but no scar and necrosis were observed. Under light microscopy, at the 2nd day, the endothelial cells were mild swollen and small vessels were embolized by PYM-AMS. At the 7th day, the endothelial cells were mild swollen. At the 14th day, the endothelial cells were proliferative and the wall of the central auricular artery had more layers. The lumen of the central artery changed mildly, while the surface of PYM-AMS was absorbed. At the 21st day, the wall of the central auricular artery was proliferative obviously and the artery became sclerostenosed, while cell division was observed. The PYM-AMS was obviously absorbed and some artery was sclerostenosed, while the wall of small vein was proliferative, too. The proliferative intima was proved to be muscle cells with Masson stain and van Gieson's stain. There were many smooth muscle cells, fibroblasts, myofibroblasts in the original blood vascular lumen on the 21st day under electron microscopy. CONCLUSION: Pingyangmycin nonspecifically made the endothelia and vessels injured, and induced proliferation of endothelial cells and smooth muscle cells, and gradually the vessels became sclerostenosed. PYM-AMS had both sustained-releasing effect and embolization effect. PYM-AMS may be a better drug for treatment of large venous malformations and arteriovenous malformations.

[An experimental study on PLGA carrier for oral tissue engineered mucosa by subcutaneous implantation in rabbits].

OBJECTIVE: To look for the best carrier for cultivating oral tissue engineered mucosa. METHODS: 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. RESULTS: 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. CONCLUSION: 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.

[Studies on the transfection of umbilical endothelia with catalytic subunit of telomerase].

OBJECTIVE: To investigate the variety of proliferating ability of umbilical endothelia (UE) transfected by plasmid pBABE-HYGR-hTERT. METHODS: 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. RESULTS: 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). CONCLUSION: The transfection of pBABE-HYGRO-hTERT had greatly improved the proliferating abilities and activated the telomerase of UE.

[Study of the relative factors of the biological behavior of pleomorphic adenoma in parotid].

OBJECTIVE: To study the growth way of parotid pleomorphic adenoma and the relative factors. METHODS: 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. RESULTS: 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. CONCLUSION: 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.