Effects of compound protein powder on nitrogen metabolism and physiological and biochemical indexes in rats / 公共卫生与预防医学

Objective To explore the nutritional efficacy of compound protein powder formulations from different sources. Methods Three groups of compound protein powder formulations were obtained through scientific blending using soy protein, whey protein and yeast protein as raw materials. The effects of the compound protein powders on nitrogen metabolism, serum biochemical indicators, and pathological changes of liver tissue and epididymal fat in rats were evaluated. Results Compared with the control (casein), the net protein utilization, biological evaluation, and protein efficacy ratio of the compound protein powders in rats were significantly improved, and the changes in these indicators in the formula with the highest whey protein content were most significant among all three formulas. The compound protein powders effectively increased the levels of albumin and globulin, while decreased the content of total cholesterol, indicating beneficial effects on improving immunity and controlling lipid metabolism, with the formula group 2 being the most effective among all three groups. The pathological examination showed that the three groups of protein powder did not have adverse effects on liver tissue and epididymal fat. Conclusion The present study demonstrates that the compound protein powder formulation has nutritional value, which suggests a potential of the application of the compound protein powder formulation in the elderly, and people with special nutritional needs, such as sports people.

Investigate mechanism of Jinzhen Oral Liquid for prevention COVID-19 based on network pharmacology and molecular docking technology / 中草药

Objective: To investigate the mechanism of Jinzhen Oral Liquid (JOL) for prevention COVID-19 through network pharmacology and molecular docking technology. Methods: The protein targets related to COVID-19 were searched by literature mining and retrieving in DisGeNET, OMIM, KEGG and UniProt databases. With the aid of Traditional Chinese Medicine Network Pharmacology Intelligent Information Platform (TCMN) searching JOL chemical components and targets, the "herb-compound-target network" was constructed using Cytoscape-3.2.1 software to predict the main active ingredients and action targets of JOL in the treatment of COVID-19. The crystal structure of novel coronavirus (SARS-CoV-2) 3CL hydrolase (3CLpro) and angiotensin converting enzyme II (ACE2) was retrieved from the RCSB PDB database, and the active compounds were docked with the two proteins by using AutoDock Vina software. Results: The herb-compound-target network contained 75 compounds including isoglabrolide, peimisine, and sennoside B, etc., which are from the three medicinal materials of Glycyrrhiza uralensis, Rheum officinale, and Fritillaria ussuriensis, and 28 targets including mammalian target of rapamycin (mTOR), Janus kinase 3 (JAK3) and mitogen-activated protein kinase 1 (MEK1). Furthermore, nine key compounds (isoglabrolide, glabrolide, ebeiedinone, desoxo- glabrolid-acetate, peimisine, verticinone, imperialine, ussuriedinone and euchrenone A5) and 10 potential targets (mTOR, JAK3, ACE2, TNFA, AKT2, PIK3CA, MEK1, BRD2, ACE and ANPEP) of JOL were predicted for treating COVID-19 by network characteristic analysis. The molecular docking results showed that some core compounds of JOL had a certain degree of affinity for 3CLpro and ACE2. Conclusion: JOL may inhibit the occurrence and development of cytokine storm in COVID-19 by regulating the expression of Brd2, CD13, and ACE2 and interfering with the PI3K/Akt, Jak-STAT, TNF and MAPK signaling pathways, and inhibit virus replication by binding with 3CLpro, thus exerting a preventive or therapeutic effect on COVID-19.

The predictive analysis of action mechanism for classic TCM excellent prescriptions of Danggui Buxue decoction on diabetes with TCMIP / 中国药理学通报

To explore the pharmacodynamic material basis and mechanism of the classic TCM excellent prescriptions(cTCMep) Danggui Buxue decoction, so as to provide more choices for the prevention and treatment of diabetes mellitus (DM). Methods With the integrative pharmacology of traditional Chinese medicine (TCMIP) , we predicted the pharmacodynamic material basis and mechanisms of the cTCMep Danggui Buxue decoction for treatment of DM. Results The 50 " Astragalus-Angelica" components in the cTCMep Danggui Buxue decoction directly or indirectly acted on 60 key drug targets. These drug targets restored the disease imbalance network of DM and its complications by acting on 30 major regulatory pathways such as nervous-endocrine system, gap junction, hormone signaling pathways, and cardiovascular circulatory system. The main components of Astragalus membranaceus membranaceus involved in the prevention and treatment of DM were astragalosides, astragalosides and alavonoids. The main components involved in the prevention and treatment of DM in Angelica sinensis were organic acids such as ferulic acid, oil components such as (3-sweet myrrh, stigmasterol-|3-D-glucoside, smbelliferol and scopolamine. Conclusions The 50 " Astragalus-Angelica" components in the cTCMep Danggui Buxue decoction may play a role in preventing and treating DM through the AVPR1A, AVPR1B and AVPR2 (AVPR: Arginine vasopressin receptor, three subtypes), of which Astragalus membranaceu is the key component.

A fast adaptive active contour model based on local gray difference for parotid duct / 南方医科大学学报

OBJECTIVE@#To establish a fast adaptive active contour model based on local gray difference for parotid duct image segmentation.@*METHODS@#On the basis of the LBF model, we added the mean difference of the local gray scale inside and outside the contour as the energy term of the driving evolution curve, and the local gray-scale variance difference was used to replace and as the control term of the energy parameter value. Two local similarity factors of different neighborhood sizes were introduced to correct the effects of image gray unevenness and boundary blur to improve the segmentation efficiency.@*RESULTS@#During image segmentation, this algorithm allowed for adaptive adjustment of the evolution direction, velocity and the energy weight of the internal and external regions according to the difference of gray mean and variance between the internal and external regions. This algorithm was also capable of detecting the actual boundary in a complex gradient boundary region, thus enabling the evolution curve to approach the target boundary quickly and accurately.@*CONCLUSIONS@#The proposed algorithm is superior to the existing segmentation algorithms and allows fast and accurate segmentation of the parotid duct with well-preserved image details.

Effects of ginkgolide K on platelet aggregation activity and neuroprotection / 中国中药杂志

To investigate the antagonism effects of different concentrations of ginkgolide K(GK) on platelet activating factor (PAF)-induced platelet aggregation and neuroprotective effect on cells and animal models of ischemia-reperfusion injury. GK-containing serum in rabbit was prepared, and the effects of GK-containing serum on PAF-induced platelet aggregation was observed by platelet aggregation assay. The effect of different concentrations of GK on apoptosis of SH-SY5Y cells injured by oxygen-glucose deprivation/reoxygenation (OGD/R) was investigated by Hoechst 33342/PI double staining in OGD/R cell model. The focal cerebral ischemia-reperfusion model (I/R)was established in rats to detect the effects of GK on neurobehavioral scores and cerebral infarction volume. GK could inhibit PAF-induced platelet aggregation, reverse the apoptosis induced by OGD/R injury and improve the neurobehavioral score and cerebral infarction volume after cerebral ischemia-reperfusion injury in rats in a dose-dependent manner. GK can inhibit PAF-induced platelet aggregation and improve nerve injury after cerebral ischemia-reperfusion.

Study on molecular mechanism of Paeoniae Rubra Radix and Phellodendri Cortex intervening Alzheimer's disease using network pharmacology methods / 中草药

Objective: To conduct computing network pharmacological studies on Paeoniae Rubra Radix (Chishao) and Phellodendri Cortex (Huangbai), and to explore their mechanism for intervening Alzheimer's disease (AD). Methods: The interactions among 199 compounds from the two kinds of Chinese Herbs (Chishao and Huangbai) and 23 approved drug targets related to AD were studied with molecular docking and network pharmacological analysis methods. Results: The most of the compounds in Chishao and Huangbai exhibit good drug-like properties. The mechanism of Chishao and Huangbai may be that they modulate the expression of GSK-3β, caspase-7, BchE, and mTOR to resist AD. Conclusion: The method of network pharmacological studies is helpful to explore the possible active molecules in Chishao and Huangbai and elucidate the mechanism of action.

A single nucleotide polymorphism in the Epstein-Barr virus genome is strongly associated with a high risk of nasopharyngeal carcinoma / 癌症

<p><b>BACKGROUND</b>Epstein-Barr virus (EBV) commonly infects the general population and has been associated with nasopharyngeal carcinoma (NPC), which has a high incidence in certain regions. This study aimed to address how EBV variations contribute to the risk of NPC.</p><p><b>METHODS</b>Using logistic regression analysis and based on the sequence variations at EBV-encoded RPMS1, a multi-stage association study was conducted to identify EBV variations associated with NPC risk. A protein degradation assay was performed to characterize the functional relevance of the RPMS1 variations.</p><p><b>RESULTS</b>Based on EBV-encoded RPMS1 variations, a single nucleotide polymorphism (SNP) in the EBV genome (locus 155391: G>A, named G155391A) was associated with NPC in 157 cases and 319 healthy controls from an NPC endemic region in South China [P < 0.001, odds ratio (OR) = 4.47, 95% confidence interval (CI) 2.71-7.37]. The results were further validated in three independent cohorts from the NPC endemic region (P < 0.001, OR = 5.20, 95% CI 3.18-8.50 in 168 cases vs. 241 controls, and P < 0.001, OR = 5.27, 95% CI 4.06-6.85 in 726 cases vs. 880 controls) and a non-endemic region (P < 0.001, OR = 7.52, 95% CI 3.69-15.32 in 58 cases vs. 612 controls). The combined analysis in 1109 cases and 2052 controls revealed that the SNP G155391A was strongly associated with NPC (P(combined) < 0.001, OR = 5.27, 95% CI 4.31-6.44). Moreover, the frequency of the SNP G155391A was associated with NPC incidence but was not associated with the incidences of other EBV-related malignancies. Furthermore, the protein degradation assay showed that this SNP decreased the degradation of the oncogenic RPMS1 protein.</p><p><b>CONCLUSIONS</b>Our study identified an EBV variation specifically and significantly associated with a high risk of NPC. These findings provide insights into the pathogenesis of NPC and strategies for prevention.</p>

Oncogenic induction of cellular high CpG methylation by Epstein-Barr virus in malignant epithelial cells / 癌症

Epstein-Barr virus (EBV) is a well-known human herpesvirus associated with virtually all nasopharyngeal carcinoma (NPC) and approximately 10% of gastric cancer (GC) worldwide. Increasing evidence shows that acquired genetic and epigenetic alterations lead to the initiation and progression of NPC and GC. However, even deep whole exome sequencing studies showed a relatively low frequency of gene mutations in NPC and EBV-associated GC (EBVaGC), suggesting a predominant role of epigenetic abnormities, especially promoter CpG methylation, in the pathogenesis of NPC and EBVaGC. High frequencies of promoter methylation of tumor suppressor genes (TSGs) have been frequently reported in NPC and EBVaGC, with several EBV-induced methylated TSGs identified. Further characterization of the epigenomes (genome-wide CpG methylation profile--methylome) of NPC and EBVaGC shows that these EBV-associated tumors display a unique high CpG methylation epigenotype with more extensive gene methylation accumulation, indicating that EBV acts as a direct epigenetic driver for these cancers. Mechanistically, oncogenic modulation of cellular CpG methylation machinery, such as DNA methyltransferases (DNMTs), by EBV-encoded viral proteins accounts for the EBV-induced high CpG methylation epigenotype in NPC and EBVaGC. Thus, uncovering the EBV-associated unique epigenotype of NPC and EBVaGC would provide new insight into the molecular pathogenesis of these unique EBV-associated tumors and further help to develop pharmacologic strategies targeting cellular methylation machinery in these malignancies.

Aberrant promoter CpG methylation and its translational applications in breast cancer / 癌症

Breast cancer is a complex disease driven by multiple factors including both genetic and epigenetic alterations. Recent studies revealed that abnormal gene expression induced by epigenetic changes, including aberrant promoter methylation and histone modification, plays a critical role in human breast carcinogenesis. Silencing of tumor suppressor genes (TSGs) by promoter CpG methylation facilitates cells growth and survival advantages and further results in tumor initiation and progression, thus directly contributing to breast tumorigenesis. Usually, aberrant promoter methylation of TSGs, which can be reversed by pharmacological reagents, occurs at the early stage of tumorigenesis and therefore may serve as a potential tumor marker for early diagnosis and therapeutic targeting of breast cancer. In this review, we summarize the epigenetic changes of multiple TSGs involved in breast pathogenesis and their potential clinical applications as tumor markers for early detection and treatment of breast cancer.

Promoter methylation of tumor suppressor genes in esophageal squamous cell carcinoma / 癌症

Esophageal squamous cell carcinoma (ESCC) is a prevalent and fatal cancer in China and other Asian countries. Epigenetic silencing of key tumor suppressor genes (TSGs) is critical to ESCC initiation and progression. Recently, many novel TSGs silenced by promoter methylation have been identified in ESCC, and these genes further serve as potential tumor markers for high-risk group stratification, early detection, and prognosis prediction. This review summarizes recent discoveries on aberrant promoter methylation of TSGs in ESCC, providing better understanding of the role of disrupted epigenetic regulation in tumorigenesis and insight into diagnostic and prognostic biomarkers for this malignancy.

Epigenetic disruption of cell signaling in nasopharyngeal carcinoma / 癌症

Nasopharyngeal carcinoma (NPC) is a malignancy with remarkable ethnic and geographic distribution in southern China and Southeast Asia. Alternative to genetic changes, aberrant epigenetic events disrupt multiple genes involved in cell signaling pathways through DNA methylation of promoter CpG islands and/or histone modifications. These epigenetic alterations grant cell growth advantage and contribute to the initiation and progression of NPC. In this review, we summarize the epigenetic deregulation of cell signaling in NPC tumorigenesis and highlight the importance of identifying epigenetic cell signaling regulators in NPC research. Developing pharmacologic strategies to reverse the epigenetic-silencing of cell signaling regulators might thus be useful to NPC prevention and therapy.

Protective effect of Annexin 5 on human sperm membrane and DNA integrity / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the role of Annexin 5 in protecting human sperm membrane and DNA integrity.</p><p><b>METHODS</b>We collected 53 semen samples based on the criteria of sperm density > 20 x 10(6)/ml and motility > 60%, and divided them into an experimental group (2.5 microl 10(-6) mol/L Annexin 5 added to 47.5 microl semen), a negative control group (2.5 microl 1 mol/L Tris-HCl [pH 8.0, 25 degrees C] added to 47.5 microl semen), and a blank control group (2.5 microl 0.01 mol/L PBS [pH 7.4] added to 47.5 microl semen). After 20 minutes of incubation, we evaluated the sperm membrane integrity using the hypoosmotic swelling test and, after another 60 minutes of treatment with H2O2 at 2.5 microl 10.02 mol/L, measured the sperm nuclear DNA integrity by acridine orange fluorescent staining.</p><p><b>RESULTS</b>After 20 minutes of treatment with Annexin 5, the experimental group showed extremely significant difference in the percentage of hypoosmotic swelling sperm ([66.17 +/- 12.02] %) from the blank control ([58.13 +/- 13.08]%, P < 0.01) and the negative control group ([59.94 +/- 11.91]%, P < 0.01), but there was no significant difference between the latter two. Treatment with H2O2 remarkably increased DFI in the experimental group (6.39 +/- 1.07) as compared with the blank control (11.16 +/- 1.16) and the negative control group (10.86 +/- 1.05, P < 0.01), but no significant difference was observed between the latter two.</p><p><b>CONCLUSION</b>Annexin 5 can increase the percentage of hypoosmotic swelling sperm in vitro and protect sperm membrane integrity, and it can also protect sperm DNA from H2O2 damage.</p>

Survey of quality of life in the patients with revision endoscopic sinus surgery / 临床耳鼻咽喉头颈外科杂志

OBJECTIVE@#To explore the effect of revision endoscopic sinus surgery (RESS) on quality of life in the patients with chronic rhinosinusitis with nasal polyposis (CRSwNP).@*METHOD@#To survey and evaluate 60 cases of RESS patients (treatment group) and 120 normal ones with physical examination (Control group) through the medical outcomes survey short form questions (MOS SF-36) and the Sino-Nasal Outcome Test-20 (SNOT-20), and comparison and analysis of the two groups results which we got were carried out.@*RESULT@#With SF-36 scales for evaluation of quality of life, the results show that: the scores of CRSwNP patients (treatment group) without surgical treatments with RESS were significantly lower than that of the control group(P 0.05).@*CONCLUSION@#RESS may obviously improve the clinical symptom of CRSNP patients. The SF-36 and SNOT-20 assessment scales could reflect the patient's QoL change.

Suppression to ameloblastoma xenografts of chicken embryo chorioallantoic membrane by tissue inhibitor of metalloproteinases-2 / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To explore the invasiveness of xenografts on chicken embryo chorioallantoic membrane (CAM) after tissue inhibitor of metalloproteinase-2 (TIMP-2) gene transfection.</p><p><b>METHODS</b>Fresh ameloblastoma tissues were minced into 1-2 mm3 and transplanted on the CAM. There were three groups named as control group (Empt), plasma transfection group (Lipo), and TIMP-2 gene transfection group (P). The specimens were respectively investigated by microscope indifferent spots after implanting. The volume of the xenografts and the weight of xenografts in the termination time of the experiment were recorded. The invasiveness of xenografts was divided into four grades by pathological examination. Western blot analysis was performed to investigate matrix metalloproteinase-2 (MIMP-2) and TIMP-2 protein in xenografts.</p><p><b>RESULTS</b>Ameloblastoma tissues can survive on CAM and the tumor cells may invade it on 5-7 days after implanting. At 9 d after implanting, the invasiveness grades in P group were 7 in grade 0, 1 in grade 2, 0 in grade 3. The expression of TIMP-2 protein in P group was significantly higher than that in Empt group (P < 0.05). The expression of MMP-2 protein in P group was lower than that in Empt group (P < 0.05).</p><p><b>CONCLUSION</b>The xenotransplanted tumor model of human ameloblastoma on CAM was successfully established. The invasiveness of ameloblastoma xenografts was suppressed might be due to TIMP-2 gene transfection.</p>

Effects of GnRH analogues on MAPK pathway in rat Leydig cells / 中华男科学杂志

<p><b>OBJECTIVE</b>To investigate the effects of GnRH analogues GnRHa and GnRHant on the MAPK pathway in rat Leydig cells.</p><p><b>METHODS</b>Rat Leydig cells were primarily cultured for 24 hours in vitro and serum-starved for 2 hours, followed by treatment with GnRHa (10(-7) mol/L) or GnRHant (10(-6) mol/L) for 0, 5, 15, 30, 60 and 90 minutes, with the 0 min group as the control. Then the protein levels of phosphorylated ERK (p-ERK) and phosphorylated p38 (p-p38) were detected by Western blot, and that of p-ERK determined by the same means after co-incubation of GnRHa or GnRHant with the PKC inhibitor GF109203X at 1, 5, 10 and 20 micromol/L.</p><p><b>RESULTS</b>After stimulation of the Leydig cells with GnRHa or GnRHant for different times, the protein level of p-p38 showed no significant difference from that of the control group (P > 0.05). Then the Leydig cells were treated with GF109203X at different concentrations for 20 minutes and with addition of GnRHa for another 10 minutes. The level of p-ERK was significantly decreased (P < 0.05) by GF109203X at 10 and 20 micromol/L. Compared with the control, the p-ERK expression was increased by 65% at 15 minutes (P < 0.05) in the GnRHant stimulation group, by 81% (to the peak) at 30 minutes (P < 0.05), began to fall at 60 minutes, and returned to the base level at 90 minutes. The p-ERK level exhibited no significant difference from that of the control (P > 0.05) after treatment of the Leydig cells with different concentrations of GF109203X for 20 minutes and then with GnRHant for 30 minutes.</p><p><b>CONCLUSION</b>The ERK MAPK activation induced by GnRHa depends on the PKC pathway, but not that induced by GnRHant. The p-38 MAPK pathway may not be involved in the effect of GnRH analogues on rat Leydig cells.</p>

Expression of rat annexin 5 and its effect on human sperm motility in vitro / 中华男科学杂志

<p><b>OBJECTIVE</b>Gonadotropin releasing hormones (GnRH) regulate the expression of annexin 5 in Leydig cells, and annexin 5 is supposed to be a signal molecule in regulating testosterone secretion. This study aimed to investigate the function of annexin 5 in male reproduction by observing its effect on human sperm motility in vitro.</p><p><b>METHODS</b>The encoding sequence of rat annexin 5 was chemically synthesized and inserted into the HIS fusion expression vector pET28a. The expression of the fusion protein HIS-annexin 5 was induced by isopropyl-beta-D-thiogalactoside (IPTG) under the control of the T7 promoter, and the products were purified by affinity chromatography. The anticoagulant activity of annexin 5 was determined by the modified activated partial thromboplastin time (APTT) test. Semen samples from 15 donors were assigned to a control and an annexin 5 group, the latter treated with recombinant annexin 5 at the concentration of 10(-8) mol/L. Sperm motility and the percentage of grade a + b sperm were measured by computer-assisted semen analysis (CASA) after 20 and 60 min exposure, and the sperm ascending experiment was done after 20 min treatment.</p><p><b>RESULTS</b>The product of the synthesized target gene was 947 bp in length, and the inserted sequence corresponded to the published encoding sequence of rat annexin 5. The plasmid pET28a-annexin 5 was transformed into E. coli BL21(DE3) and IPTG induced a fusion protein with a relative molecular weight of about 36,000, a purity of 95% and a high anticoagulant activity. Compared with the control group, sperm motility and the percentage of grade a + b sperm were increased by 40% (P < 0.01) and 21% (P < 0.01), respectively, after 20 min treatment with annexin 5, but neither showed any significant improvement after 60 min. The sperm ascending altitude was remarkably elevated after annexin 5 treatment, with extremely significant difference from the control group (37.84 +/- 6.35 vs. 49.5 +/- 12.27, P < 0.01).</p><p><b>CONCLUSION</b>An annexin 5 recombinant expression vector was successfully constructed. The protein annexin 5 can be efficiently expressed in E. coli and effectively improve human sperm motility in vitro.</p>

Survey of quality of life in the patients with revision endoscopic sinus surgery / 临床耳鼻咽喉头颈外科杂志

Objective:To explore the effect of revision endoscopic sinus surgery (RESS) on quality of life in the patients with chronic rhinosinusitis with nasal polypsis (CRSwNP).Method:To survey and evaluate 60 cases of RESS patiens (treatment group) and 120 normal ones with physical examination (Control group) through the medical outcomes survey short form questions (MOS SF-36) and the Sino-Nasal Outcome Test-20 (SNOT-20), and comparison and analysis of the two groups results which we got were carried out.Result:With SF-36 scales for evaluation of quality of life, the results show that: the scores of CRSwNP patients (treatment group) without surgical treatments with RESS were significantly lower than that of the control group(P0.05 ).Conclusion:RESS may obviously improve the clinical symptom of CRSNP patients. The SF-36 and SNOT-20 assessment scales could reflect the patient's QoL change.

Treatment of infantile hemangioma in parotid gland region with pingyangmycin injection / 中华医学美学美容杂志

Objective To evaluate the clinical efficacy of pingyangmycin (PYM) injection on infantile hemangioma located in the parotid gland region. Methods Twelve patients were treated by intralesional injection of PYM. When necessary, the injections were repeated at an interval of one week, but not more than 3-4 sessions within a therapeutic period. Normally, the secondary therapeutic period was performed 1 month later. The general and local adverse responses were recorded and the clinical outcomes were assessed with a follow-up of 1 to 3 years. Results Complete clinical resolutions were achieved in 10 patients. 2 patients received one injection, 3 patients received 2 injections, 3 patients received 3 injections, and 2 patients received 4 injections. The remaining 2 patients with partial resolution received 6 and 7 injections respectively. No clinical recurrence was observed during the follow-up of 1 to 3 years. No ulcerations or postoperative sears in injection regions were presented. The function of facial nerve was remained normality in all patients. The systematic side effects included transient pyrexia and poor appetite appeared in partial patients. No allergy cases were found. Conclusion Treatment of infantile hemangioma located in parotid gland region with PYM injection reveals a high rate of complete clinical resolution, with fair cosmetic results and short treatment time, and it does not damage the facial nerve or form local scar.The treatment time of PYM injection seems to be positively related to size of the lesions.

Relationship between lymphangiogenesis in colorectal cancer and metastasis using the D2-40 antibodies / 中国综合临床

Objective To investigate the clinical pathologic significance of lymphangiogenesis in in colorectal cancer. Methods New lymphatic-specific markers D2-40 was used immunohistochemically to detect the lymphatic vessel density(LVD) in the intratumoural and peritumoral areas, and in normal tissue from 96 cases of colorectal cancer, which were analyzed with clinical pathologic parameters of those colorectal cancer. Results Significandy higher LVD was found in the intratumoural area(14.5±2.4), when compared with normal(5.9±1.1)and peritumoural areas(6.7±1.2) (P<0.01). LVD of the peritumoural area was higher than normal area (P< 0.01). However, peritumoural LVD was associated with both depth of invasion and liver metastasis (r=0.71,0.78 P<0.05), but not associated with tumour size, macroscopic type and lymph-node metastasis (P>0.05). Intratu-moural LVD was not correlated with tumour size, macroscopic type, the depth of invasion,lymph-node metastasis, and liver metastasis(P>0.05). Conclusion Lymphangiogenesis in the peritumoural area may be helpful in evalution of liver metastasis and prognosis.

Characterization of growth and proliferation in a telomerase-immortalized ameloblastoma cell line / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To establish and characterize the cell line of ameloblastoma (AM) by transfection with human telomerase reverse transcriptase (hTERT).</p><p><b>METHODS</b>Primary cultures of AM cells were infected with a retroviral vector encoding hTERT. Infected cells were selected and checked by immunocytochemistry (ICC), in vitro proliferation, reverse transcriptase polymerase chain reaction (RT-PCR), senescence associated beta galactosidase staining (SA-beta-Gal staining), telomerase activity assay.</p><p><b>RESULTS</b>Compared to the uninfected cells, which arrested at the population doublings (PDL) of 6, the infected cells were more active in proliferation and reached 65 PDL to date. ICC confirmed the epithelial origin of the infected cells based on positive pan-cytokeratin and negative vimentin expression. There was no senescent signal in infected cells but not in uninfected cells. hTERT mRNA and telomerase activity were detected stably in infected cells.</p><p><b>CONCLUSIONS</b>The infected AM cells were immortalized after transfection with hTERT and can serve as a genetically defined model for AM study.</p>