Study on therapeutic mechanism of Rehmanniae Radix Praeparata- Corni Fructus in sequelae of ischemic stroke based on network pharmacology technology / 中国中药杂志

In ischemic stroke sequela phase, Rehmanniae Radix Praeparata-Corni Fructus drug pair has the effect in protecting damaged neurons, but its mechanism has not been clear. In this study, network pharmacology was used to predict the mechanism of Rehmanniae Radix Praeparata-Corni Fructus in the treatment of ischemic stroke sequela. Through database search and literature retrie-val, 40 active ingredients of Rehmanniae Radix Praeparata and Corni Fructus were obtained, and their targets were obtained through STITCH and TCMSP databases. The targets of ischemic stroke sequela were obtained through OMIM,GAD,TTD and DrugBank databases. By screening the intersections of active ingredients targets and stroke treatment targets, 21 potential targets were obtained. The DAVID database was used for GO enrichment analysis and KEGG pathway analysis of potential targets. GO enrichment analysis showed that Rehmanniae Radix Praeparata-Corni Fructus were mainly involved in regulation of blood pressure, negative regulation of extrinsic apoptotic signaling and positive regulation of angiogenesis. KEGG pathway analysis showed that Rehmanniae Radix Praeparata-Corni Fructus could inhibit inflammatory response and apoptosis signaling pathway by regulating HIF-VEGFA signaling pathway in neural stem cell proliferation, TNF signaling pathway and NF-kappaB signaling pathway. Molecular docking technique was used to verify that Rehmanniae Radix Praeparata-Corni Fructus component has a good binding activity with potential targets. The results showed that in ischemic stroke sequela phase, Rehmanniae Radix Praeparata-Corni Fructus drug pair could play an important role in recovering neural function, promoting the proliferation of neural stem cells, angiogenesis, preventing neural cells apoptosis and regulating inflammatory factors.

Role of ataxia telangiectasis mutated in the oncogenesis of oral squamous cell carcinoma / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the role of ataxia telangiectasis mutated (ATM) gene in the oncogenesis and progression of oral squamous cell carcinoma (OSCC).</p><p><b>METHODS</b>A total of 61 formalin-fixed and paraffin-embedded samples were obtained from patients with hyperkeratosis, oral leukoplakia, OSCC and normal healthy controls. The expression of ATM protein in all of the samples was investigated by streptavidin-peroxidase immunohistochemistry assay. PCR was also performed to detect the loss of heterozygosity (LOH) in D11s2179 of ATM gene. The correlations between ATM and the clinical and histopathological characteristics were also investigated.</p><p><b>RESULTS</b>The results indicated that the ATM expression was increased in oral premalignant lesions (P<0.05). For OSCC, 68.8% samples showed normal or increased ATM expression, while 31.3% had decreased or absent ATM expression. Significant differences were found between the group of decreased or absent ATM expression and that of normal or increased expression over the histopathological grade and lymph node metastasis state. PCR results displayed that none of the samples from oral premalignant lesions showed abnormal changes, while 3 of the OSCC (9.38%) showed loss of heterozygosity (LOH) and 2 (6.25%) with microsatellite instability (MSI). Those 3 samples of LOH showed absent ATM expression.</p><p><b>CONCLUSION</b>These findings indicated that the over expression of ATM may contribute to prevent carcinogenesis of OSCC. ATM inactivation may be one of the genetic alterations of the molecular progression of OSCC.</p>

Amplification methods of DNA from paraffin-embedded tissues of oral leukoplakia / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To set up a feasible method for detection of objective genes from paraffin-embedded tissues of oral leukoplakia (OLK).</p><p><b>METHODS</b>Twenty-five pieces of formalin-fixed, paraffin-embedded tissues of OSCC were selected and ATM gene was detected respectively by 3 methods: the microdissection-nested PCR method, proteinase K-PCR method and conventional phenol-chloroform-PCR method. The positivity rates were compared statistically.</p><p><b>RESULTS</b>The positivity rates of these 3 methods were 84%, 52% and 64% respectively. Significant difference was found in positivity rate between the microdissection-nested PCR method and the proteinase K-PCR method (P = 0.032).</p><p><b>CONCLUSIONS</b>The microdissection-nested PCR method merits recommendation because it is more efficient, easy to perform and has the advantage of less sample amount.</p>

Application and estimation of subjective criterion for judging immune status of recurrent aphthous ulceration patients / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To adopt a criterion summarized in long-term practice to judge the immune status of recurrent aphthous ulceration (RAU) patients, and to analyze its rationality and feasibility.</p><p><b>METHODS</b>60 cases were included, whose clinical information, such as age, sex, constitution status, history of system diseases and severity of ulceration were collected. The subjective judgement results of the immune status were obtained according to the criterion summarized in clinical practice. At the same time, peripheral blood parameters about immune function were examined in order to obtain the objective examination results of the immune status. With the examination result as the gold standard, the rationality of the criterion was estimated through its sensitivity and specificity.</p><p><b>RESULTS</b>According to subjective judgment, there were 86.67% cases with abnormal immune status (28.33% cases with elevated immune status, and 58.33% cases with reduced immune status). According to objective examination, there were 93.33% cases with abnormal immune status (23.33% cases with elevated immune status, and 70.00% cases with reduced immune status. Sensitivity and specificity were 89.29% and 50.00% when the criterion was used to judge immune abnormality, 42.86% and 76.09% when the criterion was used to judge immune elevation, and 64.29% and 55.56% when the criterion was used to judge immune reduction.</p><p><b>CONCLUSION</b>It is of some rationality and feasibility to judge the immune status according to the criterion based on clinical information, and the criterion can be used in small clinics.</p>

Correlation of oral Candida spp. and denture-wearing in healthy elderly / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the frequency and intensity of Candida spp. incidence from the oral cavities of the healthy elderly in Chengdu, and to study the role of the dentures in the distribution of oral Candida spp.</p><p><b>METHODS</b>A total of 212 individuals(age > 60 years) were divided into four groups: A1 (48 man with dentures), B1 (61 man without dentures), A2 (53 women with dentures) and B2 (50 women without dentures). Samples of their oral flora were obtained by rinsing with 10 mL PBS solution. The samples were centrifuged and resuspended in PBS (500 microL), and plated onto Sabouraud's dextrose agar. CHROMagar Candida, sugar assimilation patterns (API 20C AUX tests) were used to determine Candida spp. The total number of yeast colonies on the plates was considered as the relative intensity of oral Candida.</p><p><b>RESULTS</b>Candida spp. was isolated from 116 healthy elderly individuals (54.72%), such as C. albicans, C. parapsilosis, C. krusei, C. guilliermondii, C. tropicalis, etc. The frequency of Candida spp. in A1, B1, A2 and B2 was 66.67%, 36.07%, 64.15%, and 56.00%, respectively. The frequency of C. albicans in A1, B1, A2 and B2 was 56.25%, 21.31%, 56.60% and 38.00%, respectively. The frequency of Candida spp. and the intensity of Candida spp. were greater for individuals in the denture-wearing group than that in the control.</p><p><b>CONCLUSION</b>The frequency and intensity of Candida spp. incidence from the healthy elderly are closely correlated with denture-wearing, and the differences of the frequency and intensity of Candida spp. incidence in the elderly are due to the differences of frequency and intensity of C. albicans incidence.</p>

Analysis of chromosome karyotype of oral carcinoma-associated fibroblasts / 华西口腔医学杂志

<p><b>OBJECTIVE</b>The purpose of this study was to investigate whether the fundamental genetic character of oral carcinoma-associated fibroblasts changes through contrasting and analyzing the oral carcinoma-associated fibroblasts and the normal fibroblasts of oral mucosa.</p><p><b>METHODS</b>The two kinds of cells were treated with colchicine and microsometic fluid, and then were expanded with cold acetic acid and formalized with methyl alcohol. The cells were observed under the oil microscope after Giemsa staining. The chromosome karyotype of the two kinds of cells was analyzed by Visus 2. 1.</p><p><b>RESULTS</b>There were not obvious differences in the way of chromosome karyotype between the oral carcinoma-associated fibroblasts and the normal fibroblasts of oral mucosa.</p><p><b>CONCLUSION</b>The basic genetic characteristics of the normal cells are conserved in the oral carcinoma-associated fibroblasts, which means the cells have no malignant changes.</p>

Adhesion to buccal epithelial cells of Candida albicans isolates from oral lichen planus / 华西口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the adhesion to buccal epithelial cells of Candida albicans isolates from erosive oral lichen planus (OLP) and nonerosive OLP, and its role in the development of OLP.</p><p><b>METHODS</b>A total of 112 isolates, comprising healthy control (26), erosive OLP (62) and nonerosive OLP (24), were screened for the adhesion by using buccal epithelial cell (BEC) assay.</p><p><b>RESULTS</b>The adhesion to buccal epithelial cells of the isolates from erosive OLP group was stronger than that of those from healthy control.</p><p><b>CONCLUSION</b>Candida albicans, some isolates with a special virulence attribute may contribute to the occurrence and progression of erosive OLP.</p>

Amplification of EMS1 gene in oral carcinogenesis / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the amplification of EMS1 gene in the carcinogenesis of oral mucosa.</p><p><b>METHODS</b>A total of 78 subjects, including 30 patients with oral leukoplakia (OLK), 33 with oral squamous cell carcinoma (OSCC), and 15 healthy controls, were studied. By using microdissection method, we obtained normal mucosa, hyperplastic epithelia, mild-dysplastic epithelia, moderate-dysplastic epithelia, severe-dysplastic epithelia and primary OSCC tissue. Then we analyzed EMS1 amplification by using differential PCR.</p><p><b>RESULTS</b>EMS1 amplification began from moderate-dysplastic epithelia and occurred in 20.0% OLK cases and 57.6% OSCC cases. In the progress of OSCC, no gene amplification was observed in normal tissues, non-dysplastic OLK and mild-dysplastic OLK, while in the cases with metastasis, amplification frequency increased significantly (P = 0.015).</p><p><b>CONCLUSIONS</b>EMS1 amplification parallels with the progress of oral carcinogenesis, indicating their potential roles in oral carcinogenesis.</p>

Centrosome hyperamplificationin oral precancerous lesions and squamous cell carcinomas / 华西口腔医学杂志

<p><b>OBJECTIVE</b>It is currently considered that the defect of mitotic spindle caused by centrosome abnormalities may be one of the reasons for the development of aneuploidy in tumors. This study attempted to elucidate the possible role of centrosome defects in the development and progression of OSCC by investigating the frequency of centrosome amplification in oral precancerous lesions and OSCC.</p><p><b>METHODS</b>Formalin-fixed, paraffin-embedded tissues of 12 cases of normal oral epithelium, 22 case of dysplasia with different degree epithelium dysplasia and 32 cases of OSCC with different differentiation were investigated for centrosome status by using indirect immunofluorescence double staining with antibodies to centrosome protein gamma-tubulin and cytokeratin. The differences and the change trend of centrosome status in these groups were statistically analyzed by SPSS10.0.</p><p><b>RESULTS</b>Normal oral epithelium showed normal centrosomes in epithelium cells, while 16 of 22 cases (72.73%) of dysplasia (DYS) and 27 of 32 cases (84.38%) of OSCC showed the evidence of centrosome amplification and morphological abnormalities characterized by huge size, clump or supernumerary centrosomes in a fraction of epithelium or tumor cells. The percentage of cells with abnormal centrosomes increased gradually from mild-dysplasia epithelium to poorly differentiated OSCC, which positively correlated with the histologicalcytologic grade of oral precancerous lesions and OSCC (P < 0.01).</p><p><b>CONCLUSION</b>Centrosome amplification was an early event and that might play a role in the establishment and perhaps the progression of OSCC. There might be some direct relationship between centrosome defects and the cellular morphological phenotype characteristics of dysplasia and OSCC. Centrosome amplification could be served as an alternative diagnostic indicator of dysplasia and the intervention of centrosome cycle might serve as a particular way for the prevention and treatment of OSCC in the future.</p>

Human oral carcinoma-associated fibroblasts: its cultivation, purification and identification / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To separate, cultivate, purify and identify oral carcinoma-associated fibroblasts (CAFs) preliminarily.</p><p><b>METHODS</b>The primary oral CAFs and normal fibrolasts (NFs) of oral mucosa were obtained by tissue culture. Then cells were dissociated by 0.25% trypsin and purified by curettage method and trypsinization. Morphological characteristics were observed under light microscope and electron microscope. The certain proteins were examined by immunohistochemistry (SP method).</p><p><b>RESULTS</b>The third passage purified oral CAFs were maintained. The characteristics of shape and growth of the oral CAFs changed significantly comparing to the NFs. Myofilament and electron dense patch were showed in the oral CAFs by electron microscope. The oral CAFs showed negative staining for cytokeratin, and positive staining for vimentin, alpha-smooth muscle actin and matrix metalloproteinases-2.</p><p><b>CONCLUSIONS</b>There are obvious differences of the morphological characteristics and expression of certain proteins between the CAFs and NFs. The microecology of the oral tumor-host interface might be one of the most important factors affecting the CAFs.</p>

The attribute of Candida albicans isolates from patients with oral lichen planus / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To investigate the genotypic profiles of Candida albicans isolates from erosive oral lichen planus (OLP) and nonerosive OLP, and then to compare the results with their virulence attributes.</p><p><b>METHODS</b>A total of 112 isolates from healthy control (26), erosive OLP (62) and nonerosive OLP (24) were screened for genotypic profiles by using the randomly amplified polymorphic DNA (RAPD) assay. In addition, adhesion to buccal epithelial cells assay and phospholipase activity assay were used to evaluate the virulence attributes of these isolates.</p><p><b>RESULTS</b>RAPD analyses with some random primer revealed 4 different genotypes among all isolates, and there was significant difference in the geneotypic constitution between every two groups. Statistically, in healthy group the major type was B and D, however, the major type in erosive OLP was A and C, and the major type in nonerosive OLP was A and D. The isolates with genotype A had the strongest adherence among 4 genotypes. The phospholipase activity of the isolates with genotype A and C were higher than that with genotype B and D.</p><p><b>CONCLUSIONS</b>Some Candida albicans isolates with special genotypic profiles and virulence attributes may contribute to the development and progression of OLP.</p>

Effect of millimeter therapy in burning mouth syndrome / 中华口腔医学杂志

<p><b>OBJECTIVE</b>To observe the millimeter wave therapy responses in patients with burning mouth syndrome.</p><p><b>METHODS</b>Eighty patients were randomized divided into 4 groups. The first group was treated with both millimeter wave irradiation and routine medication, the second group with millimeter wave irradiation, the third with pretending millimeter wave irradiation and routine medication and the fourth with routine medication. Pain, extravasated blood level and autonomic nerve system condition were double-blindly evaluated either before or after the treatment.</p><p><b>RESULTS</b>Statistically significant difference (P < 0.05) was found as the degree of pain was compared before and after treatment of all the 4 groups. The first and second group, which were affected by the millimeter wave irradiation, had obvious improvements in the extravasated blood level and autonomic nerve system condition (P < 0.05). When the 4 groups were compared with each other, there were significant differences (P < 0.05) between the first and the fourth groups, and the second and the fourth groups regarding the reduction of pain. According to extravasated blood level, significant differences (P < 0.05) were found in the first and third or fourth groups, the second and third or fourth groups.</p><p><b>CONCLUSIONS</b>The irradiation of holographic point by millimeter wave can improve the patients' pain, extravasated blood level and autonomic nerve system condition. It might provide a new treatment method for burning mouth syndrome.</p>