Four New Species and One New Record of Thelephora from China.

Species of the genus Thelephora (Thelephorales, Thelephoraceae) are ectomycorrhizal symbionts of coniferous and broad-leaved plants, and some of them are well-known edible mushrooms, making it an exceptionally important group ecologically and economically. However, the diversity of the species from China has not been fully elucidated. In this study, we conducted a phylogenetic analysis based on the internal transcribed spacer (ITS) regions, using Maximum Likelihood and Bayesian analyses, along with morphological observations of this genus. Four new species from China are proposed, viz., T. dactyliophora, T. lacunosa, T. petaloides, and T. pinnatifida. In addition, T. sikkimensis originally described from India is reported for the first time from China. Thelephora dactyliophora, T. pinnatifida, and T. sikkimensis are distributed in subtropical forests and mainly associated with plants of the families Fagaceae and Pinaceae. Thelephora lacunosa and T. petaloides are distributed in tropical to subtropical forests. Thelephora lacunosa is mainly associated with plants of the families Fagaceae and Pinaceae, while T. petaloides is mainly associated with plants of the family Fagaceae. Line drawings of microstructures, color pictures of fresh basidiomes, and detailed descriptions of these five species are provided.

Identification of immunological bioprocesses involved in peri-implantitis using weighted gene co-expression network analysis.

BACKGROUND: Peri-implantitis is an irreversible infectious disease that occurs with high incidence. Exploring the immune responses of peri-implantitis is key to developing targeted treatment strategies. However, there is limited research on the immune response of peri-implantitis. METHODS: This study performed a weighted gene co-expression network analysis to identify the peri-implantitis related gene network and conducted a functional enrichment analysis of the gene network. Thereafter, the candidate hub genes were selected by constructing a protein-protein interaction network and drawing an upset plot. The hub genes were identified through their significant associations with disease condition and validated using quantitative reverse transcription-polymerase chain reaction (qRT-PCR) analysis. Using the gene set variation analysis, the hub genes were further used to explore infiltrating immunocytes and immune factors in peri-implantitis. Finally, the immunocytes and immune factor related hub genes were intersected to obtain the therapeutic target, which was validated using histological staining. RESULTS: The peri-implantitis related gene network was enriched in innate and adaptive immune response. Subsequently, interleukin (IL)1B, IL10, ITGAM, ITGB1, STAT3, and TLR4 were identified as hub genes. Plasmacytoid dendritic cells, macrophages, myeloid-derived suppressor cells, natural killer T cells, and immature B cells were positively and significantly related to the hub genes IL1B, TLR4, ITGAM, and ITGB1 (correlation coefficient > 0.80). While immune factors CXCL10, IL6, and CXCL12 and hub genes IL10 and IL1B held the highest degree in the immune factors network. IL1B may be a promising therapeutic target. CONCLUSION: This study provides new insights into the hub genes, immunocytes, and immune factors underlying peri-implantitis immunological bioprocess.

Osteotomy combined with the trephine technique for invisible implant fracture: A case report.

BACKGROUND: Implant fracture is one of the most serious mechanical complications of dental implants. Conventional treatment necessitates visibility of the apical portion of the fractured implant, whereas for deep and invisible implant fractures, the traditional trephine method has been ineffective. Surgical removal of the marginal bone to expose the fracture surface would be a time-consuming and extensively damaging procedure. Here, we propose a novel technique to address invisible implant fractures. CASE SUMMARY: A 50-year-old woman was referred to our department with the chief complaint that her right mandibular implant tooth had fallen out 3 mo earlier. Cone-beam computed tomography examination showed an implant fracture with a fracture surface 5.1 mm below the crestal ridge. The patient was treated with osteotomy combined with the trephine technique to expose the surgical field and remove the implant. The invisible fractured implant was successfully removed, with minimal trauma. A modified Wafer technique-supported guided bone regeneration treatment was then administered to restore the buccal bone wall and preserve the bone mass. Six months later, fine regenerative bone and a wide alveolar crest in the edentulous area were observed, and a new implant was placed. Four months later, restoration was completed using a cemented ceramic prosthesis. Clinical and radiographic examinations 12 mo after loading fulfilled the success criteria. The patient reported no complaints and was satisfied. CONCLUSION: Osteotomy combined with the trephine technique can be effectively used to address deep and invisible implant fractures.

Dynamic navigation system-guided trans-inferior alveolar nerve implant placement in the atrophic posterior mandible: A case report.

BACKGROUND: In atrophic posterior mandibular areas, where the bone height superior to the inferior alveolar nerve (IAN) is less than 6 mm, short implants are not applicable. Conventional alternatives such as IAN transposition and various alveolar bone augmentation approaches are technically demanding and prone to complications. CASE SUMMARY: Computer-guided dynamic navigation implantation improves the accuracy, predictability, and safety of implant placement. This case report presents a dynamic navigation system-guided trans-IAN implant placement technique, which can successfully treat a posterior mandibular dentition defect when the bone height is only 4.5 mm. The implant was inserted into the buccal side of the IAN and was 1.7 mm away from the IAN. The implantation deviations were controlled within a satisfying range, and the long-term restoration outcome was stable. CONCLUSION: Dynamic navigation system-guided trans-IAN implant placement might be a recommended technique for patients with extremely insufficient residual bone height and sufficient bone width in the posterior mandibular area.

FMRP-absence-induced up-regulation of hypothalamic MAP1B expression decreases AgRP level linking with reduces in food intake and body weight.

Fragile X mental retardation protein (FMRP), strongly associated with fragile X syndrome, plays important roles by regulating gene expression via interacting with other RNA binding proteins in the brain. However, the role of FMRP in hypothalamus, a central part responsible for metabolic control, is poorly known. Our study shows that FMRP is primarily located in the hypothalamic arcuate nucleus (ARC). Using proteomic analysis, we identified 56 up-regulated and 22 down-regulated proteins in the hypothalamus of Map1b KO mice, with microtubule-associated protein 1 B (MAP1B) being the most outstanding increased protein (more than 10 folds). Immunofluorescent assays showed that MAP1B significantly increased in the Map1b-KO ARC, in which the number of agouti-related peptide (AgRP)-staining neurons significantly reduced, but not altered for pro-opiomelanocortin (POMC) neurons. We further showed an age-dependent reduces in food intake and body weight of the KO mice, along with the decreases of MAP1B and AgRP at the same time points. In hypothalamic GT1-7 cells, the AgRP expression decreased upon knockdown of FMRP or overexpression of MAP1B, and increased in response to overexpression of FMRP or knockdown of MAP1B. Co-knockdown or co-overexpression of FMRP and MAP1B led to a reverse expression of AgRP compared to overexpression of knockdown of FMRP alone, demonstrating that MAP1B is essential for the regulatory effect of FMRP on AgRP expression. Taken together, these data suggest that FMRP-deficiency-induced increase of hypothalamic MAP1B and decrease of AgRP might be associated with reduces in food intake and body weight.

Two bone blocks sandwich technique for horizontal reconstruction of severely atrophic alveolar ridge in anterior maxilla: A case report.

BACKGROUND: Severe horizontal bone deficiency of the maxillary anterior region is considered a major challenge in reconstruction and successful implant placement. Various approaches have been developed to augment bone volume. Of these approaches, onlay bone graft, alveolar bone splitting, and guided bone regeneration have been suggested. CASE SUMMARY: A 22-year-old female patient, with no previous medical history, presented to the Department of Oral Implantology, Wuhan University due to a missing right maxillary incisor. The X-ray results showed severe horizontal bone deficiency, with an available bone width of 3.1-4.0 mm. The two bone blocks sandwich technique was performed to augment the bone volume. After 6 months healing, X-ray results showed that the newly formed alveolar ridge dimension increased to 4.7-9.5 mm horizontally. Implant insertion surgery was performed and all-ceramic restorations were fabricated. The implant was stable at the 1-year follow-up visit after restoration, and the X-ray showed a stable bone level around the dental implant. The scores for the pink esthetic score and white esthetic score were 12 and 8, respectively, and the patient was satisfied with the esthetic outcome. CONCLUSION: The two bone blocks sandwich technique may be an alternative treatment option in augmenting severe horizontal bone deficiency of the anterior maxilla.

[Rev-erbß Knock-out Affectsthe Proliferation, Migration and Invasion Ability of Hepatocellular Carcinoma HepG2 Cell Line in vitro].

OBJECTIVE: To investigate the effect of nuclear receptor Rev-erbß knockout on proliferation and migration ability of human hepatocellular carcinoma cell line HepG2. METHODS: -The Rev-erbß gene knockout HepG2 cell line was abtained by CRISPR/Cas9 genome editing technique with specific DNA modification of the target gene. The Rev-erbß gene targeting vectors were co-transfected into HepG2 cells. Through cloning and screening, the Rev-erbß gene knockout HepG2 cell line was constructed, PCR, sequencing and Western blot methods were carried out for the identification of the Rev-erbß gene knockout HepG2 cell line. The expression level of tumor migration and invasion-associated gene in Rev-erbß gene knockout cell was determined by real-time quantitative PCR (qRT-PCR) and was compared with normal cell as control.MTT, cell scratch and Transwell experiments were conducted in order to explore the effect of Rev-erbß gene on HepG2 cell's ability of proliferation, migration and invasion. RESULTS: A Rev-erbß gene knockout monoclonal cell line, which was identified by PCR, sequencing and Western blot, was successfully constructed and named HepG2 C5 (Rev-erbß -/-). qRT-PCR results showed that Rev-erbß knockout resulted in up-regulation of matrix metalloproteinase-2 (MMP2), matrix metalloproteinase-9 (MMP9) and extracellular matrix protein-1 (ECM1) gene expression (P < 0.05) and down-regulation of E-cadherin (CDH1) gene expression (P=0.05).Results of MTT, cell scratch and transwell experiments showed that HepG2 C5 had stronger proliferation, migration and invasion ability than control cells (P < 0.05). CONCLUSION: Rev-erbß gene knockout could change the expression of migration and adhesion-associated genes in HepG2 cell, and then affect the proliferation, migration and invasion ability of HepG2 cells.

Lentiviral-Mediated Overexpression of MicroRNA-141 Promotes Cell Proliferation and Inhibits Apoptosis in Human Esophageal Squamous Cell Carcinoma.

BACKGROUND: Esophageal Carcinoma (EC) is the eighth most common cancer worldwide. Numerous studies have highlighted a vital role of microRNAs (miRNAs) in the development of EC. However, the mechanism of microRNA (miRNA)-141 in Esophageal Squamous Cell Carcinoma (ESCC) remains unknown. OBJECTIVE: In this study, we explored the effects of miRNA-141 on EC cell proliferation, apoptosis, xenograft tumour growth and their possible mechanisms. METHODS: A lentivirus-vector-expressing miRNA-141 was constructed, and a TE-1 cell line of ESCC with a stable expression of miRNA-141 was transfected and screened. The miRNA-141 expression level was detected using qRT-PCR. Effects of miRNA-141 overexpression on cell proliferation and apoptosis were detected using MTT and flow cytometry, respectively. Using a dual-luciferase reporter assay, a direct interaction between miRNA-141 and the 3'-Untranslated Region (UTR) of YAP1 and SOX17 was confirmed. Tumour xenograft experiment in nude mice was used to detect the tumour growth, and the effects of miRNA-141 overexpression on YAP1 and SOX17 were analysed using Western blot. RESULTS: We found that miRNA-141 was highly expressed in TE-1 cells, and miRNA-141 overexpression promoted cell proliferation and inhibited apoptosis. Moreover, the miRNA-141 group showed significantly increased tumour growth ability, luciferase activities and expression levels of YAP1 and SOX17 in the miRNA-141group were significantly down-regulated. CONCLUSION: miRNA-141 promotes cell proliferation and inhibits apoptosis in ESCC by downregulating the expression level of YAP1 and SOX17, indicating that miRNA-141 may be a potential molecular target for the treatment of ESCC.

[Establishment of a bioluminescent MDA-MB-231 cell line for in vivo imaging of human triple-negative breast cancer xenograft].

OBJECTIVE: To establish a bioluminescent MDA-MB-231 cell line which can stably express luciferase and green fluorescent protein to allow bioluminescent imaging in nude mouse models bearing human triple-negative breast cancer xenografts. METHODS: The lentivirus carrying luc2, eGFP and neo fusion genes were packaged in 293T cells via calcium phosphate co-precipitation. Human triple-negative breast cancer cell line MDA-MB-231 was infected by the lentivirus, and the positive cell clones were tested for eGFP and luc2 expressions by fluorescence microscopy and Xenogen IVIS200 bioluminescent imaging system, respectively. MTT assay, transwell invasion assay and wound healing assay were performed to evaluate the changes in the proliferation, invasion and migration abilities of the infected cells. The cells were then orthotopically implanted into the right second mammary fat pat of female BALB/c nude mice. The tumor growth was monitored by the in vivo imaging system every week, and the tumor tissues were harvested to evaluate the in vivo stability and tumorigenicity of the modified cells using cryosection and HE staining. RESULTS: The lentivirus-infected MDA-MB-231cells could stably express luc2 and eGFP, and the luciferase activity reached 9689 phontons/s/per cell. No significant changes occurred in the biological activities of the lentivirus-infected MDA-MB-231 cells. We successfully established the nude mouse model bearing orthotopically implanted human triple-negative breast cancer cells. CONCLUSION: The modified MDA-MB-231 cell line can be detected sensitively at the primary implantation site and distant metastasis site in nude mice, which provides a convenient and sensitive platform for the research of metastatic mechanism and new antitumor drugs of human triple-negative breast cancer. The combination of eGFP and luc2 is superior to single reporter gene.

[Study on construction of chimeric adenovirus vector Ad5/11 carrying human eGFP and endostatin-K5 and its experimental investigation in vitro].

AIM: To construct chimeric adenoviral vector Ad5/11 carrying reporter gene eGFP and human endostatin-K5. METHODS: Chimeric adenoviral backbone vector expressing eGFP was generated by overlap PCR and homologous recombination in E.coli BJ5183. Then chimeric adenoviral vector Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP carrying eGFP and human endostatin-K5 was constructed by co-transfecting Pac I linearized chimeric adenoviral backbone and adenoviral E1 shuttle vector expressing human endostatin-K5 into HEK 293 cells. The expression of eGFP was observed under fluorescent microscope. The expression of human endostain-K5 in U87MG cells infected by chimeric adenoviral vector was detected by RT-PCR. The infection efficiency between chimeric adenovirus and unmodified control adenovirus for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231 in vitro was evaluated by the comparison of the expression of eGFP. RESULTS: Chimeric adenovirus Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP could successfully express eGFP and endostatin-K5. Chimeric adenoviral vector significantly enhances the infection efficiency for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231 compared with unmodified adenoviral vector Ad5 E1-CMV-eGFP. CONCLUSION: Chimeric adenoviral vector Ad5/11-E1-CMV-endo-K5/E3-CMV-eGFP can significantly improve the infection efficiency for human glioblastoma cell line A172 and breast cancer cell line MDA-MB-231.

[Construction and expression of recombinant adenovirus vector carrying human endostatin, K5 and endostatin-K5 gene].

AIM: To construct the recombinant adenoviral vectors expressing human endostatin, K5 and endostatin-K5 gene respectively, and study their bioactivity in vitro. METHODS: Human endostatin, K5 and endostatin-K5 gene were amplified by PCR, which were then subcloned into shuttle vector pAd5-CMV-H1H2-MCS-6His by enzyme and ligation respectively. The positive recombinant plasmids linearized by Pac I were cotransfected into HEK 293 cells with the Pac I linearized adenoviral backbone plasmid using calcium phosphate precipitation method. The recombinant viruses were purified by CsCl density gradient centrifugation. The protein expression at different time points (24 h, 48 h and 72 h) was determined by Western blot. The inhibitory effect of the protein on ECV-304 growth was detected by MTT. RESULTS: The recombinant adenoviral vectors expressing human endostatin, K5 and endostatin-K5 gene were successfully constructed in HEK293 cells. Protein expression was detected by Western blot. The level of protein expression was increased with the prolonged incubation of the infected HeLa cells. Three kinds of the protein expressed by the recombinant adenoviral vectors showed obvious inhibitory effect on ECV-304 cell growth. CONCLUSION: The protein expressed by adenoviral vectors carrying endostatin, K5 and endostatin-K5 gene has an obvious inhibitory effect on ECV-304 cell growth.

[Effect of crestal rotated flap on esthetics of peri-implant soft tissues in anterior maxilla].

OBJECTIVE: To investigate the height of interdental papillae around single-tooth implants in the anterior maxillae after exposure of implant using crestal rotated flap. METHODS: The study comprised 34 implants in the anterior maxillae from 32 patients. All implants were uncovered by use of crestal rotated flap technique. Impressions were taken before and after stage 2 surgery, and stone models were used to evaluate the change of papillae height. The gingival papillae index around single-tooth implants was compared before and after the surgery. RESULTS: The average increase in papillae height was (0.77 +/- 0.25) mm, which was statistically significant with analysis of a paired t test (P < 0.05). The change of gingival papillae index showed no significant difference using statistical analysis of rank sum test (P > 0.05). The lower the gingival papillae index was before the surgery, the less the papillae height increased after the surgery. CONCLUSIONS: The main advantages of the crestal rotated flap technique are simplicity and predictability, and it consistently provides high papillae for the maxillary implants.

[Influence of implant-abutment connection mode on stress distribution in peri-implant bone].

OBJECTIVE: To investigate the influence of butt joint connection and platform switching design of implant-abutment connection on the stress distribution in peri-implant bone. METHODS: Three-dimensional finite element models of implant-supported mandibular first molar with different implant-abutment connections were computed by COSMOSM 2.85. Traditional butt joint connection was used in model A and platform switching design in model B. Loading conditions were a vertical load of 200 N and inclined load of the same magnitude at 45 degrees to the vertical axis of the implant. Stress distribution in peri-implant bone and von Mises stresses at the same point of buccal and lingual implant-bone interfaces in two models were compared. RESULTS: Stresses concentrated in peri-implant cortical bone at the neck of implants on the buccal and lingual sides; maximum von Mises stresses under inclined load were higher than those under vertical load. Maximum von Mises stress in bone was 11.61 MPa in model A and 7.15 MPa in model B under vertical load, and 22.07 MPa in model A and 11.87 MPa in model B under inclined load respectively. Von Mises stresses decreased as the distance from implant-abutment junction increased and the most obvious change occurred at the interface between cortical bone and spongy bone. Von Mises stresses at the same points of buccal and lingual implant-bone interfaces in model A were higher than those in model B. CONCLUSIONS: Compared with butt joint connection, platform switching design improved the stress distribution and decreased the maximum stresses in peri-implant bone around implant cervix.

[Construction of an expression vector for high expression of siRNA].

AIM: To construct the vector for efficient expression of siRNA using pre-mir30 backbone. METHODS: By chemical synthesis method, pre-mir30 backbone introduced an appropriate restriction enzyme site for foreign shRNA inserting was cloned into an expressing vector containing U6 promoter. The silencing efficiency of a new siRNA expressing vector was detected by transfection and Western blot. RESULTS: The new vector containing pre-mir30 backbone expressing siRNA against GFP could markedly inhibit the expression of GFP compared with the vector expressing control siRNA. CONCLUSION: siRNA expressing vector constructed by pre-mir30 backbone could highly express foreign siRNA.