[Morphological study of sexual reproductive disorders in Ligusticum chuanxiong].

Chuanxiong Rhizoma is a well-known Sichuan-specific herbal medicine. Its original plant, Ligusticum chuanxiong, has been cultivated asexually for a long time. L. chuanxiong has sexual reproductive disorders, which restricts its germplasm innovation. However, there is little research on the reproductive system of L. chuanxiong. This study is based on a comparative anatomical research approach, using morphological dissection, paraffin sectioning, staining and compression, and combined with scanning electron microscopy technology, to observe and compare the flowers, fruits, and seeds at various stages of reproductive growth of L. chuanxiong and its wild relative L. sinense. The results showed that the meiosis of pollen mother cells is abnormal in L. chuanxiong anthers, and the size and number of microspores are uneven and inconsistent in the tetrad stage. tapetum cells are not completely degenerated during anther development. During the pollen ripening stage, there are fine cracks in the anther wall, while most anthers could not release pollen normally. The surface of mature pollen grains is concave and partially deformed, and the pollens are all inactive and cannot germinate in vitro. The starch, polysaccharides, and lipids in the pollen were insufficient. The filaments of L. chuanxiong are short at the flowering stage and recurved downward. Double-hanging fruits were observed in the fruiting stage, being wrinkled; with shriveled seeds. Compared with L. sinense at the same stage, the anthers of L. sinense developed normally, and the pollen grains are vigorous and can germinate in vitro. The double-hanging fruits of L. sinense are full and normal; at the flowering period, the filaments are long and erect, significantly higher than the stigma. Mature blastocysts are visible in the ovary of both L. chuanxiong and L. sinense, and there is no significant difference in stigmas. The conclusion is that during the development of L. chuanxiong stamens, the meiosis of pollen mother cells is abnormal, and tetrad, tapetum, filament and other pollen structures develop abnormally. L. chuanxiong has the characteristic of male infertility, which is an important reason for its sexual reproductive disorders.

Effect the accumulation of bioactive constituents of a medicinal plant (Salvia Miltiorrhiza Bge.) by arbuscular mycorrhizal fungi community.

BACKGROUND: Arbuscular mycorrhizal fungi (AMF) form symbiotic relationships with various terrestrial plants and have attracted considerable interest as biofertilizers for improving the quality and yield of medicinal plants. Despite the widespread distribution of AMFs in Salvia miltiorrhiza Bunge's roots, research on the impact of multiple AMFs on biomass and active ingredient accumulations has not been conducted. In this study, the effects of five native AMFs (Glomus formosanum, Septoglomus constrictum, Rhizophagus manihotis, Acaulospora laevis, and Ambispora gerdemannii) and twenty-six communities on the root biomass and active ingredient concentrations of S. miltiorrhiza were assessed using the total factor design method. RESULTS: Thirty-one treatment groups formed symbiotic relationships with S. miltiorrhiza based on the pot culture results, and the colonization rate ranged from 54.83% to 89.97%. AMF communities had higher colonization rates and total phenolic acid concentration than single AMF, and communities also appeared to have higher root fresh weight, dry weight, and total phenolic acid concentration than single inoculations. As AMF richness increased, there was a rising trend in root biomass and total tanshinone accumulations (ATTS), while total phenolic acid accumulations (ATP) showed a decreasing trend. This suggests that plant productivity was influenced by the AMF richness, with higher inoculation benefits observed when the communities contained three or four AMFs. Additionally, the affinities of AMF members were also connected to plant productivity. The inoculation effect of closely related AMFs within the same family, such as G. formosanum, S. constrictum, and R. manihotis, consistently yielded lower than that of mono-inoculation when any combinations were applied. The co-inoculation of S. miltiorrhiza with nearby or distant AMFs from two families, such as G. formosanum, R. manihotis, and Ac. laevis or Am. gerdemannii resulted in an increase of ATP and ATTS by more than 50%. AMF communities appear to be more beneficial to the yield of bioactive constituents than the single AMF, but overall community inoculation effects are related to the composition of AMFs and the relationship between members. CONCLUSION: This study reveals that the AMF community has great potential to improve the productivity and the accumulation of bioactive constituents in S. miltiorrhiza, indicating that it is an effective way to achieve sustainable agricultural development through using the AMF community.

Identification of core genes in prefrontal cortex and hippocampus of Alzheimer's disease based on mRNA-miRNA network.

Alzheimer's disease (AD) is a neurodegenerative disorder with cognitive impairment and abnormal mental behaviour. There is currently no effective cure. The development of early diagnostic markers and the mining of potential therapeutic targets are one of the important strategies. This study aimed to explore potential biomarkers or therapeutic targets related to AD in the hippocampus and prefrontal cortex, two brain regions highly related to AD. Differentially expressed genes and miRNAs between AD patients and healthy controls were obtained from the Gene Expression Omnibus database. The mRNA-miRNA network was constructed and key genes involved in AD were screened out by protein-protein interaction analysis, and were subsequently verified by independent datasets and qPCR in an AD mouse model. Our findings showed that six hub genes including CALN1, TRPM7, ATR, SOCS3, MOB3A and OGDH were believed to be involved in the pathogenesis of AD. Western blot analysis further determined that CALN1, ATR and OGDH were the possible biomarkers and therapeutic targets for AD. In addition, 6 possible miRNAs biomarkers have also been verified by qPCR on AD animal models. Our findings may benefit clinical diagnosis and early prevention of AD.

Immediate acupuncture with GB34 for biliary colic: protocol for a randomised controlled neuroimaging trial.

INTRODUCTION: As the main manifestation of gallstone disease, biliary colic (BC) is an episodic attack that brings patients severe pain in the right upper abdominal quadrant. Although acupuncture has been documented with significance to lead to pain relief, the immediate analgesia of acupuncture for BC still needs to be verified, and the underlying mechanism has yet to be covered. Therefore, this trial aims first to verify the immediate pain-alleviation characteristic of acupuncture for BC, then to explore its influence on the peripheral sensitised acupoint and central brain activity. METHODS AND ANALYSIS: This is a randomised controlled, paralleled clinical trial, with patients and outcome assessors blinded. Seventy-two patients with gallbladder stone disease presenting with BC will be randomised into a verum acupuncture group and the sham acupuncture group. Both groups will receive one session of immediate acupuncture treatment. Improvements in patients' BC will be evaluated by the Numeric Rating Scale, and the pain threshold of acupoints will also be detected before and after treatment. During treatment, brain neural activity will be monitored with functional near-infrared spectroscopy (fNIRS), and the needle sensation will be rated. Clinical and fNIRS data will be analysed, respectively, to validate the acupuncture effect, and correlation analysis will be conducted to investigate the relationship between pain relief and peripheral-cerebral functional changes. ETHICS AND DISSEMINATION: This trial has been approved by the institutional review boards and ethics committees of the First Teaching Hospital of Chengdu University of Traditional Chinese Medicine, with the ethical approval identifier 2019 KL-029, and the institutional review boards and ethics committees of the First People's Hospital of Longquanyi District, with the ethical approval identifier AF-KY-2020071. The results of this trial will be disseminated through peer-reviewed publications and conference abstracts or posters. TRIAL REGISTRATION NUMBER: CTR2000034432.

[Microecology and geoherbalism of traditional Chinese medicine].

Microecology was directly or indirectly involved in the growth and development, metabolism process, and component accumulation of traditional Chinese medicine(TCM) in various ways, which affected the formation and changes of the geoherbalism of TCM. It was one of the main tasks of traditional Chinese medical microecology(TCMM) to reveal the relationship among microecological structure and its change rule and the quality effect of TCM. The heterogeneity of soil environment caused by geographical and climatic factors, as well as the discreteness limitation caused by isolation factors such as distance and host selection, were the main causes of the differentiation of microecological geography of TCM. The microecology of TCM had important influences and contributions on the distinctive origin and quality of Dao-di herbs, which was mainly reflected in the formation of excellent germplasm(including disease and insect resistance, drought resistance, salt resistance, cold resistance, etc.), the increase of yield, the formation of medicinal parts, the metabolism and accumulation of effective components, the time limit of harvesting, and the toxicity, increasing efficiency or reducing toxicity of TCM in the processing, the changes of product efficiency after introduction, and the authenticity of fungus medicine. With the vigorous development of metabonomics and modern information technology, the following aspects would become the future research trends, including the microecologically mediated biogenic pathway of chemical components, the metabolic synthesis reactor of TCM based on the microecological quantitative effect relationship, the cultivation of genuine Chinese medicine based on reconstruction of microecological structure, the origin identification barcode traceability technology, and the toxicity reduction and efficiency enhancement technology of TCM based on the microecological.

Global clonal spread of mcr-3-carrying MDR ST34 Salmonella enterica serotype Typhimurium and monophasic 1,4,[5],12:i:- variants from clinical isolates.

OBJECTIVES: To investigate the prevalence and transmission of mcr-3 among Salmonella enterica serotype Typhimurium and 1,4,[5],12:i:-. METHODS: A total of 4724 clinical Salmonella isolates were screened for the presence of mcr-3 in China during 2014-19. The clonal relationship of the mcr-3-positive isolates and their plasmid contents and complete sequence were also characterized based on WGS data from the Illumina and MinION platforms. RESULTS: We identified 10 mcr-3-positive isolates, and all were MDR, mostly resistant to colistin, cefotaxime, ciprofloxacin, doxycycline and florfenicol. mcr-3 was co-present with blaCTX-M-55-qnrS1 on hybrid ST3-IncC-FII conjugatable plasmids (n = 6) and an ST3-IncC non-conjugatable plasmid (n = 1) and embedded into a pCHL5009T-like IncFII plasmid on the Salmonella chromosome (n = 3). Four distinctive genetic contexts surrounded mcr-3 and all but one were closely related to each other and to the corresponding region of IncFII plasmid pCHL5009T. IS15DI was most likely the vehicle for integration of mcr-3-carrying IncFII plasmids into ST3-IncC plasmids and the chromosome and for shaping the MDR regions. In addition, a phylogenetic tree based on the core genome revealed a unique Salmonella lineage (≤665 SNPs) that contained these 10 mcr-3-positive isolates and another 38 (33 from patients) mcr-3-positive Salmonella from five countries. In particular, most of the 51 mcr-3-positive isolates belonged to ST34 and harboured diverse antibiotic resistance genes (ARGs), including mcr-3-blaCTX-M-55-qnrS1, and possessed similar ARG profiles. CONCLUSIONS: Our findings revealed global clonal spread of MDR ST34 Salmonella from clinical isolates co-harbouring mcr-3 with blaCTX-M-55 and qnrS1 and a flexibility of mcr-3 co-transmittance with other ARGs mediated by mobile genetic elements.

[Characteristics of hydrogen and oxygen isotopes in precipitation and moisture sources in Gaoyou, Jiangsu Province, China]. / æ±Ÿè‹é«˜é‚®å¤§æ°”é™æ°´æ°¢æ°§åŒä½ç´ ç‰¹å¾åŠæ°´æ±½æ¥æº.

It is necessary to examine the characteristics of hydrological cycle in Gaoyou area of Jiang-su Province in response to climate change and flood disasters. In this study, 121 atmospheric rain samples were collected and environmental factors were recorded from July 2015 to October 2017. We analyzed the hydrogen and oxygen stable isotopic composition of rain and identify moisture sources in this area. Results showed that the hydrogen and oxygen isotopic composition of rain had an seasonal variation, which was positive in the winter but negative in the summer. D-excess value was higher in winter than that in summer. On the annual scale, significantly negative relationships between δD (δ18O) and temperature and between δD (δ18O) and precipitation indicated the "anti-temperature effect" and "precipitation amount effect", respectively. On the seasonal scale, there was no obvious "temperature effect" but "precipitation amount effect" in autumn and winter. Results from the HYSPLIT model showed that the precipitation in this area came mainly from ocean evaporation that was influenced by the Chinese South Sea, Indian Ocean and Pacific Ocean. Precipi-tation in other seasons mainly came from the water vapor mixture from the Eurasian continent, Atlantic Ocean, Arctic Ocean, as well as local evaporation. The seasonal pattern of δD and δ18O values in precipitation was mainly influenced by monsoon activity and El Niño-Southern Oscillation (ENSO). In addition, the precipitation isotopes clearly indicated the shift in climates from El Niño to La Niña.

Increased formation of neutrophil extracellular traps is associated with gut leakage in patients with type 1 but not type 2 diabetes.

BACKGROUND: The aim of this study was to investigate the association of the formation of neutrophil extracellular traps (NETs) with gut leakage in type 1 (T1D) and type 2 diabetes (T2D). METHODS: In all, 105 subjects (56 T1D, 49 T2D) were included in the study. Eight biomarkers of NET formation and gut leakage (ie, protein arginine deiminase type 4 [PAD4], neutrophil elastase [NE], proteinase 3 [PR3], complement 5a [C5a], α1 -antitrypsin [AAT], DNase I, zonulin, and lipopolysaccharide [LPS]) were measured in serum samples by ELISA. Neutrophils were isolated and stimulated by phorbol myristate acetate to form NETs in vitro. Neutrophil intracellular contents were then collected and used as antigens to detect anti-neutrophil cytoplasmic antibodies (ANCA) in the serum. RESULTS: There was an increase in NET-associated proteins (PAD4, NE, PR3, C5a, AAT and DNase I) in new-onset T1D patients but not in those with T2D. Of PAD4, NE, and PR3, PAD4 was found to be the most sensitive biomarker for the diagnosis of T1D. Furthermore, circulating levels of zonulin and LPS were not only increased, but were also strongly correlated with NET formation and ANCA generation in T1D patients. CONCLUSIONS: This study provides evidence that increased formation of NETs, particularly PAD4, is closely associated with gut leakage in T1D but not T2D, and suggests that microorganisms and the release of neutrophil cytoplasmic antigen during the formation of NETs may be involved in the pathogenesis of T1D.

[Research and prospect of Traditional Chinese Medical Microecology].

In the long-term evolution, microbes and hosts coexist widely, forming a symbiotic microecosystem and resulting the complex interactions of the metabolism. With the application of microecological theory in Chinese materia medica science, two main points have been accepted gradually. On the one hand, the prevention and treatment of human diseases by traditional Chinese medicines can be achieved through the correction and adjustment of the imbalance of the human microecosystem. On the other hand, the microecosystem can regulate the quality of traditional Chinese medicines in real time, and further affect their curative effect. Thus, a new discipline, Traditional Chinese Medical Microecology, has been gradually established. In this review, the background, theoretical structure, research directions, key problems and the relationship with human microecology of Traditional Chinese Medical Microecology were systematically summarized and prospected for promoting its development. Moreover, this review provides a reference protocol for further discoursing the microecological mechanism involving the efficacy of traditional Chinese medicines.

Modified technique for preservation of inferior alveolar nerve during mandibulectomy.

BACKGROUND: The purpose of this article is to introduce the modified technique of preservation of the inferior alveolar nerve (IAN) during mandibulectomy for a benign lesion. METHODS: Five cases of osteofibrous hyperplasia and 3 cases of centricity osteomyelitis were included. During surgery, the IAN was marked using a planned cutting guide. Using an oscillating saw, the depth of the osteotomy along the IAN was controlled until the bone cortex was cut through. After splitting, the bony section was removed, leaving the neurovascular bundle intact. The sensation of the lower lip was evaluated using current perceptive threshold testing during follow-up. RESULTS: After follow-up for 6-27 months, no recurrence or secondary deformity was found. One patient had severe sensory disturbance. CONCLUSION: With the use of a cutting guide and osteotomy tricks, mandibulectomy with preservation of the IAN can be accurately performed.

Three-Dimensional Measurement and Cluster Analysis for Determining the Size Ranges of Chinese Temporomandibular Joint Replacement Prosthesis.

The aim of this study was to investigate the osseous characteristics of Chinese temporomandibular joint (TMJ) and detect the size clusters for total joint prostheses design.Computer tomography (CT) data from 448 Chinese adults (226 male and 222 female, aged from 20 to 83 years, mean age 39.3 years) with 896 normal TMJs were chosen from the Department of Radiology in the Shanghai 9th People's Hospital. Proplan CMF 1.4 software was used to reconstruct the skulls. Three-dimensional (3D) measurements of the TMJ fossa and condyle-ramus units with 13 parameters were performed. Size clusters for prostheses design were determined by hierarchical cluster analyses, nonhierarchical (K-means) cluster analysis, and discriminant analysis.The glenoid fossa was grouped into 3 clusters, and the condyle-ramus units were grouped into 4 clusters. Discriminant analyses were capable of correctly classifying 97.24% of the glenoid fossa and 94.98% of the condyle-ramus units. The means and standard deviations for the parameter values in each cluster were determined.Fossa depth and angles between the condyle and ramus were important parameters for Chinese TMJ prostheses design. 3D measurements and cluster analysis of the osseous morphology of the TMJ provided an anatomical reference and identified the dimensions of the minimum numbers of prosthesis sizes required for Chinese TMJ replacement.

[Isolation and Identification of Antagonistic Endophytic Actinomycetes Against Root Rot Disease in Ligusticum chuanxiong].

Objective: To get an understanding of the microflora of endophytic actinomycetes in Ligusticum chuanxiong,and to obtain the resource of antagonistic strains against Ligusticum chuanxiong root rot disease. Methods: Actinomycetes in stem nodes and rhizomes of Ligusticum chuanxiong were isolated and purified by a series of means,namely tissue block method, homogenate technique and plat streaking method. Further, dual culture and inhibition zone method were employed to test the antagonistic activity of these strains. To identify strains with potential precisely,both microscopic observation and DNA sequencing were conducted. Results: 83 strains of Ligusticum chuanxiong endophytic actinomycetes were identified, which consisted of 13 species groups. Among all of them,species identified as Streptomyces scopuliridis( KF600747. 1), Streptomyces griseorubiginosus( AB706352. 1) and Streptomyces agglomeratus( LC055413. 1) showed antagonistic activities against four kinds of pathogenic fungi of Ligusticum chuanxiong root rot disease. Conclusion: Actinomycetes which belong to Griseorubroviolaceus groups of streptomyce are potential biocontrol microbes to Ligusticum chuanxiong root rot,showing importance to the production of Chuanxiong Rhizoma.

[Identification of Pathogens Causing Root Rot Disease on Ligusticum chuanxiong in Sichuan].

OBJECTIVE: To identify the pathogens causing root rot disease on Ligusticum chuanxiong from the main producing areas in Sichuan, and to provide gist of management of this disease in the future. METHODS: The diseased rhizomes of Ligusticurn chuanxiong with root rot symptom were sampled in two main producing areas in Sichuan. According to Koch's postulate, the pathogenicity on Ligusticum chuanxiong rhizomes of the isolates was confirmed. The representative isolates were identified based on their morphological characteristics and ribosomal DNA internal transcribed spacer(rDNA-ITS) sequences. RESULTS: The isolates(f2-16, 3-2, f4-19 and f5-7)which caused root rot disease held high homology reached 99%, 99%, 100% and 100% respectively, when compared with GenBank sequences GQ229075, JQ796755, JN232136 and FJ481024. CONCLUSION: The causing agents of root rot disease on Ligusticum chuanxiong belong to Fusarium solani, Fusarium oxysporum, Plectosphaerella cucumerina and Phoma glomerata. Pathogens from different regions are varied.

The gene expression patterns of BMPR2, EP300, TGFß2, and TNFAIP3 in B-Lymphoma cells.

OBJECTIVE: The results of a previous study showed that a clear dysregulation was evident in the global gene expression of the BCL11A-suppressed B-lymphoma cells. In this study, the bone morphogenetic protein receptor, type II (BMPR2), E1A binding protein p300 (EP300), transforming growth factor-ß2 (TGFß2), and tumor necrosis factor, and alpha-induced protein 3 (TNFAIP3) gene expression patterns in B-cell malignancies were studied. METHODS: The relative expression levels of BMPR2, EP300, TGFß2, and TNFAIP3 mRNA in B-lymphoma cell lines, myeloid cell lines, as well as in cells from healthy volunteers, were determined by real-time quantitative reverse transcript-polymerase chain reaction (qRT-PCR) with SYBR Green Dye. Glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was used as reference. RESULTS: The expression level of TGFß2 mRNA in B-lymphoma cell lines was significantly higher than those in the cells from the healthy control (P<0.05). However, the expression level of TNFAIP3 mRNA in B-malignant cells was significantly lower than that of the healthy control (P<0.05). The expression levels of BMPR2 and EP300 mRNA showed no significant difference between B-malignant cell lines and the healthy group (P>0.05). In B-lymphoma cell lines, correlation analyses revealed that the expression of BMPR2 and TNFAIP3 (r=0.882, P=0.04) had significant positive relation. The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in cell lines from myeloid leukemia were significantly lower than those in the cells from the healthy control (P<0.05). The expression levels of TGFß2 mRNA showed no significant difference between myeloid leukemia cell lines and the healthy control or B-malignant cell lines (P>0.05). The expression levels of BMPR2, EP300, and TNFAIP3 mRNA in B-lymphoma cells were significantly higher than those of the myeloid leukemia cells (P<0.05). CONCLUSION: Different expression patterns of BMPR2, EP300, TGFß2, and TNFAIP3 genes in B-lymphoma cells exist.

Four osteotomy methods with piezosurgery to remove complicated mandibular third molars: a retrospective study.

PURPOSE: Piezosurgery has been used widely in oral and maxillofacial surgery, but there has been no report systematically describing an osteotomy method with piezosurgery for complicated mandibular third molar removal. The aim of this study was to introduce 4 osteotomy methods using piezosurgery and evaluate their effects. MATERIALS AND METHODS: A retrospective study was conducted of patients with a complicated impacted mandibular third molar requiring extraction. The predictor variable was the extraction technique. Four osteotomy methods using piezosurgery were tested according to different impaction types: method 1 involved complete bone removal; method 2 involved segmental bone removal; method 3 involved bone removal combined with tooth splitting; and method 4 involved block bone removal. Outcome variables were success rate, operative time, major complications (including nerve injury, mandible fracture, severe hematoma, or severe edema), and serious pyogenic infection. Data were analyzed using descriptive statistics. RESULTS: The study was composed of 55 patients with 74 complicated impacted mandibular third molars. All impacted mandibular third molars were removed successfully. The average surgical time was 15 minutes (range, 8 to 26 minutes). Thirty-eight molars (51.4%) were extracted by method 1, 18 molars (24.3%) by method 2, 12 molars (16.2%) by method 3, and 6 molars (8.1%) by method 4. Two cases (2.7%) developed postoperative infections and recovered within 1 week using drainage and antibiotic administration. CONCLUSION: The 4 osteotomy methods with piezosurgery provide effective ways of removing complicated impacted mandibular third molars.

Local resection of the mass to treat the osteochondroma of the mandibular condyle: indications and different methods with 38-case series.

BACKGROUND: Local resection of the mass was reported to treat the condylar osteochodroma in some cases. The purpose of this study was to evaluate the feasibility and the indications of the local resection. METHODS: In all, 47 patients with osteochondroma of the mandibular condyle were treated from January 2002 to March 2012. The decision to perform local resection depended on 2 factors: there was a stalk existing between the mass and the condyle, and the condylar surface was involved less than 1/2. RESULTS: Local resection of the mass was performed in 38 cases (80.1%). These masses were removed by 4 methods: direct removal (18 cases), pushed-out by a screw and steel wire (14 cases), excision in multiple pieces (3 cases), and temporary osteotomy of the zygomatic arch (3 cases). In the follow-up period, there was no recurrence. CONCLUSION: Local resection of the mass was a more conservative procedure to treat the solitary osteochondroma with a stalk.

[Analysis of endophytic fungi community of Ligusticum chuanxiong using PCR-DGGE].

OBJECTIVE: To research the diversity of endophytic fungal communities among Ligusticum chuanxiong growing at 5 areas in Sichuan province, and illuminate the developing mechanism of geoherbs from the microecological perspective. METHOD: The PCR-DGGE and DNA sequencing techniques were used to analyze the endophytic fungi community of L. chuanxiong. RESULT: The community of endophytic fungi present difference among different growing areas. Though minor difference were found among individuals at the same area, similarity among individuals from the same growing areas were higher significantly than those from different growing areas. Compared with the other 4 growing areas, L. chuanxiong from Shiyang town, Dujiangyan city had more abundant endophytic fungi and low similarity to others, and which probably had special types of fungi. CONCLUSION: The abundant and stable endophytic fungal community is an important factor for the development of geoherb L. chuanxiong at Shiyang town, Dujiangyan city.

[Changing of TSG101 expression on the invasion and metastasis of gastric cancer cells].

OBJECTIVE: To explore the functions of tumor susceptibility gene 101 (TSG101) in the invasion and metastasis of gastric cancer cells by cell culture. METHODS: The TSG101 eukaryotic expression and empty plasmids were transfected into gastric cancer cell line SGC7901. After screening with G418, single cell clone was selected and cultured. The expression of TSG101 was detected by reverse transcription (RT)-PCR and Western blotting. Cells were divided into TSG101 eukaryotic expression plasmid and blank control groups. Then the relationship was examined between TSG101 expression and tumor invasion and metastasis through the invasion, mobile, adhesion and damage scar experiment. RESULTS: The expression levels of TSG101 in mRNA and protein in the TSG101 eukaryotic expression group were significantly higher than those of the plasmid and blank control groups (0.85 ± 0.09 vs 0.55 ± 0.07, 0.45 ± 0.07 and 29.4 ± 1.2 vs 17.0 ± 0.4, 15.9 ± 0.4, all P < 0.05). The cell number of TSG101 eukaryotic expression group through Matrigel, laminin, type IV collagen protein (84 ± 14, 128 ± 10, 62 ± 7) were significantly higher than those of the plasmid group (55 ± 9, 77 ± 10, 31 ± 6) and blank control group (48 ± 8, 76 ± 9, 24 ± 5, all P < 0.01). The number of cells adherent to Matrigel, laminin, type IV collagen protein of the TSG101 eukaryotic expression group (0.97 ± 0.04, 1.34 ± 0.04, 0.90 ± 0.01) were obviously higher than those of the plasmid group (0.53 ± 0.03, 0.75 ± 0.05, 0.42 ± 0.02) and blank control group (0.60 ± 0.03, 0.72 ± 0.03, 0.40 ± 0.01, all P < 0.01). The number of TSG101 eukaryotic expression group cell migrating to membrane lower surface was obviously higher than that of the plasmid group and blank control group (87 ± 13 vs 54 ± 8, 48 ± 7, all P < 0.01). The fusion speed of the TSG101 eukaryotic expression group was faster than that of plasmid and blank control groups after cultivating for 24 and 48 h. CONCLUSIONS: TSG101 expression increases significantly in SGC-7901 cells after a stable transfection of TSG101 eukaryotic expression plasmids. Also the capacities of invasion and metastasis become markedly enhanced.

Computer-assisted local resection for exostosis osteochondroma of the mandibular condyle.

The purpose of this study was to evaluate the guiding effect of computer-assisted design technique in local resection of exostosis osteochondroma (EOC) from the mandibular condyle. Eight patients diagnosed with EOC through computed tomographic scan were selected from January 2011 to March 2012. SurgiCase CMF 5.0.0.32 software was used for preoperative design. The osteotomy angle, depth, and tumor shape were measured to guide the surgery. The tumors were resected from the stalk 2 mm within the normal tissue to restore the unaffected condyle as much as possible. Postoperative computed tomographic scans showed that the tumors had been resected completely in all patients. The mean (SD) error between the design and the result was 1.82 (1.25) mm. Five patients had more than 1-year follow-up without recurrence. Computer-assisted design is a good way to help local resection of EOC. It can improve the accuracy of tumor resection and keep the unaffected condyle as much as possible for function.

Combined transfection of Bcl-2 siRNA and miR-15a oligonucleotides enhanced methotrexate-induced apoptosis in Raji cells.

OBJECTIVE: B-cell lymphoma 2 (Bcl-2) is an important member of the Bcl-2 family of proteins that regulate the induction of apoptosis. This study aims to investigate whether Bcl-2 small interfering RNA (siRNA) combined with miR-15a oligonucleotides (ODN) could enhance methotrexate (MTX)-induced apoptosis in Raji cells. METHODS: Chemically synthesized miR-15a ODN and Bcl-2 siRNA were transfected in Raji cells by using a HiPerFect Transfection Reagent and then combined with MTX. Expression levels of Bcl-2 protein were detected by Western blot. Cell proliferation was determined by CCK8 assay. The rate of cell apoptosis was determined by Annexin V/PI double staining. The morphology of apoptotic cells was observed by Hoechst-33 258 staining. RESULTS: After the cells were transfected with miR-15a ODN combined with Bcl-2 siRNA, Bcl-2 protein levels were evidently decreased. CCK8 assay showed that cell proliferation was significantly decreased and was significantly lower in miR-15a ODN combined with Bcl-2 siRNA plus MTX group than in miR-15a ODN with methotrexate group, Bcl-2 siRNA with MTX group, and single MTX group (P<0.05). Hoechst 33258 staining revealed numerous apoptotic cells. AnnexinV/PI double staining showed that the apoptotic rates were (13.13±1.60)%, (34.47±2.96)%, (32.87±3.48)%, and (45.47±2.16)% in MTX, Bcl-2 siRNA plus MTX, miR-15a ODN plus MTX, and miR-15a ODN combined with Bcl-2 siRNA plus MTX groups, respectively. Among these groups, the apoptotic rate of miR-15a ODN combined with Bcl-2 siRNA plus MTX group was the highest; this apoptotic rate was also significantly different from that of miR-15a ODN or Bcl-2 siRNA plus MTX (P<0.05). CONCLUSIONS: Bcl-2 siRNA combined with miR-15a ODN could enhance MTX-induced apoptosis in Raji cells. Bcl-2 siRNA and miR-15a combined with MTX may be a useful approach to improve the treatment effects on lymphoma.