Objective To investigate the efficiency of target gene transfection of the heart and liver after tail vein or intramyocardial injection of adenovirus vector (GFP-Ad).Methods GFP-AD was constructed at first.A total of 20 male 8-week old C57BL/6 mice were randomly and equally divided into tail vein injection of GFP-AD group and intramyocardial injection of GFP-AD group.The mRNA levels of GFP in the heart and liver tissues were detected by Q-PCR at different time points.Fluorescence microscopy was performed to visualize the expression of GFP fluorescence.Results Compared with the tail vein injection group, the GFP mRNA level in mouse heart tissue was apparently higher in the intramyocardial injection group.In both groups, the GFP mRNA levels in liver tissue were significantly increased compared with that in the heart tissue.In the tail vein injection group, the GFP mRNA level in liver tissue reached a peak on day 7;but in the intramyocardial injection group, the mRNA level of GFP in liver tissue reached apeak on day 3.We also observed the same trend of GFP fluorescence expression in the tail vein injection group compared with that in the intramyocardial injection group.Conclusions Intramyocardial injection of adenovirus vector is suitable to achieve a higher transfection efficiency in mouse heart tissue compared with the tail vein injection method.Although both injection methods are suitable for transfection of mouse liver, the tail vein injection method is preferential for it is simple and less invasive.

Objective To provide the endoscopic anatomic basis and anatomic parameters for endoscopic surgical therapy on orbital lesions , and to analyze the advantages and key points of this surgical approach .Methods Five fresh adult heads were used in this experiment .Endonasal thanslamina approach and endoscopic technique were applied to observe important anatomic marks while intraoperative medicalization of the medical rectus muscle was applied to observe the exposure and positioning of important structures and trends of the optic canal and intra orbit .Results Uncinate process was at the lower front corner of middle nasal concha; ethmoidalis bulla was behind the uncinate process , and ethmoidei sinus was reachable after an incision was made on ethmoidalis bulla;anterior ethmoidal artery and posterior ethmoidal artery were the important anatomic landmarks of the inner ethmoidei sinus;optic canal prominence , carotid artery prominence and OCR were the important anatomic landmarks of the inner sphenoid sinus; lamina papyracea was at the lateral wall of ethmoidei sinus , and orbital contents were approacchable after lamina papyracea was cut off ;inside orbit , the optic nerve was approachable through the gap between the medial rectus muscle and inferior rectus muscle .The ophthalmic artery of 9 out of 10 sides of the specimens was originated from the supraclinoidal segment of the internal carotid artery while the remaining one was originated from the cavernous segment of internal carotid artery .There were 7 sides in which ophthalmic artery was located at the inferior lateral of the optic nerve;there were 2 sides in which ophthalmic artery was located at the inferior of the optic nerve; the remaining one was located at the inferior medial of the optic nerve .Conclusion The endoscopic endonasal thanslamina approach can sufficiently expose the optic nerve and the structures in the medical space of the orbit.Uncinate process, ethmoid bulla, anterior ethmoidal artery, posterior ethmoidal artery and posterior ethmoid sinus are the important landmarks of the endoscopic endonasal thanslamina approach .Optic canal prominence , internal carotid artery prominence and OCR are the important landmarks for optic canal decompression .Ophthalmic artery , orbital branches , anterior ethmoidal artery , posterior ethmoidal artery , internal carotid artery are the important vessels . Medialization of the medial rectus muscle is effective to approach the orbital anatomical structures .

Objective:To investigate the effect of lncRNA UCA1 on the radiosensitivity of in vitro cultured glioma cell lines SHG-44, U87 and U251 by regulating the miR-873-5p expression. Methods:The survival of glioma cells SHG-44, U87 and U251 treated with different radiation intensities (0, 2, 4, 6 and 8 Gy) was detected by colony formation assay. The expression levels of UCA1 in glioma cells SHG-44, U87 and U251 were measured by qRT-PCR. The radiation-resistant glioma cells U87 and U251 were selected for subsequent study. After silencing UCA1 expression and/or over-expressing miR-873-5p, the cell survival rate was detected by colony formation assay, and the cell apoptosis rate was determined by flow cytometry. The dual luciferase reporter gene assay and qRT-PCR were employed to verify the targeting relationship between UCA1 and miR-873-5p.Results:UCA1 was up-regulated in the radiation-resistant U87 and U251 cells. Silencing UCA1 or over-expressing miR-873-5p inhibited the survival of U87 and U251 cells, and promoted the cell apoptosis induced by radiation exposure. miR-873-5p was a target gene of UCA1, and UCA1 negatively regulated the expression of miR-873-5p. The inhibition of miR-873-5p could reverse the effect of silencing UCA1 on the radiosensitivity of glioma cells. Silencing UCA1 increased the inhibitory effect of radiation on the glioma cell U251 xenografts.Conclusion:Silencing UCA1 inhibits the survival of glioma cells and promotes the cell apoptosis by up-regulating the expression of miR-873-5p, thereby increasing the radiosensitivity of glioma cells.

Objective:To investigate the role of circular RNA (circRNA) circSEPT9 in the radioresistance of glioma and its molecular mechanism.Methods:Pathological samples were collected from 40 glioma patients who underwent surgery and postoperative radiotherapy in the First Affiliated Hospital of Zhengzhou University from January 2019 to January 2022. All patients were divided into the radiation-sensitive and radiation-resistant groups. The expression levels of circSEPT9 were assessed using real-time reverse transcription PCR (RT-qPCR) in two groups. The radiation-resistant glioma cell U251R was constructed based on human glioma cell U251. The U251R cells were divided into the negative control (si-NC), circSEPT9 knockdown (si-circSEPT9), negative control combined with irradiation (si-NC+4 Gy), circSEPT9 knockdown combined with irradiation (si-circSEPT9+4 Gy), circSEPT9 knockdown combined with control inhibitor and irradiation (si-circSEPT9+NC inhibitor+4 Gy), and circSEPT9 knockdown combined with miR-432-5p inhibitor and irradiation (si-circSEPT9+miR-432-5p inhibitor+4 Gy) groups. The targeting relationship between circSEPT9 and miR-432-5p was verified through dual-luciferase reporter assay. Colony formation assay was employed to assess the survival rate of U251R cells. Flow cytometry was adopted to measure the apoptosis rate. The expression level of circSEPT9 in glioma tissues was statistically analyzed using independent sample t-test. The survival and apoptosis rates in each group were evaluated using one-way ANOVA. Results:The expression level of circSEPT9 was up-regulated in the glioma tissues of patients in the radiation-resistant group (1.00±0.18 vs. 3.25±0.13, P<0.05). Compared to the si-NC group, the U251R cells in the si-circSEPT9 group exhibited a significant reduction in survival fraction and a notable increase in apoptosis rate (9.24±0.83 vs. 19.36±2.13, both P<0.05). After radiation exposure at 4, 6 and 8 Gy, si-circSEPT9 treatment significantly decreased the survival fraction in U251R cells (all P<0.05). Compared with the si-NC+4 Gy group, the apoptosis rate was increased in the si-circSEPT9+4 Gy group (18.83±1.94 vs. 35.23±3.56, P<0.05). Dual-luciferase reporter assay showed that circSEPT9 could target and negatively regulate the expression level of miR-432-5p. Compared with the si-circSEPT9+NC inhibitor+4 Gy group, the survival fraction of U251R was significantly increased in the si-circSEPT9+miR-432-5p inhibitor+4 Gy group ( P<0.05). Conclusion:Knockdown of circSEPT9 enhances the radiosensitivity of glioma cells by regulating cell apoptosis through targeting the miR-432-5p.

Objective : To explore the role and molecular mechanism of Supervillin in zebrafish embryo development. Methods : Clustal Omega and DNAman software were used to analyze and compare the homology of amino acid sequences of Svil and SVIL in zebrafish and human. The expression pattern of Svila (subtype of Svil protein in zebrafish) during early embryonic development was analyzed by RT⁃PCR and in situ hybridization. Svila expression in zebrafish was inhibited by injecting morphinos ( MO) , and morphological changes of embryos were observed. The expression and nuclear localization of β ⁃catenin protein were detected by Western blot , and the expression of Wnt/β⁃catenin target gene was detected by RT⁃PCR. Results : During the early embryonic development of zebrafish , Svila was maternally expressed and showed an upward trend with the development process. The expression of Svila was reduced by MO , and the development of zebrafish embryos was distorted and the body axis was bent , which might be related to the blocked movement of concentrated extension of embryos. Further studies showed that Svila expression affected β⁃catenin nuclear transport and Wnt/β⁃catenin signaling pathway activation. Conclusion Svila regulates the concentrated extension movement of zebrafish embryos by activating the Wnt/β⁃catenin signaling pathway.

Dental Caries is a kind of chronic oral disease that greatly threaten human being's health. Though dentists and researchers struggled for decades to combat this oral disease, the incidence and prevalence of dental caries remain quite high. Therefore, improving the disease management is a key issue for the whole population and life cycle management of dental caries. So clinical difficulty assessment system of caries prevention and management is established based on dental caries diagnosis and classification. Dentists should perform oral examination and establish dental records at each visit. When treatment plan is made on the base of caries risk assessment and carious lesion activity, we need to work out patient‑centered and personalized treatment planning to regain oral microecological balance, to control caries progression and to restore the structure and function of the carious teeth. And the follow-up visits are made based on personalized caries management. This expert consensus mainly discusses caries risk assessment, caries treatment difficulty assessment and dental caries treatment plan, which are the most important parts of caries management in the whole life cycle.
Consensus ; Dental Care ; Dental Caries/prevention & control* ; Humans ; Prevalence