Objective:To investigate the mechanism of miR-218-5p regulating the glycolytic process in human non-small cell lung cancer A549 cells by targeting phosphodiesterase 7A(PDE7A).Methods:A549 cells were routinely cultured,and miR-218-5p mimic,mimic-NC,PDE7A overexpression plasmid(PDE7A-oe)and PDE7A control plasmid(PDE7A-NC)were transfected into A549 cells using Lipo3000,and recorded as the miR-218-5p mimic group,the mimic-NC group,the PDE7A-oe group and the PDE7A-NC group.The transfection efficiency was verified by qPCR assay;the expressions of glycolysis key enzyme proteins were detected by WB assay;the 2-deoxyglucose and lactate contents in A549 cells of each transfection group were detected by glucose assay and lactate production assay;the target binding relationship between miR-218-5p and PDE7A was verified by dual-luciferase reporter gene assay,and the data from the TCGA database were used to analyze the expression level of PDE7A mRNA in lung cancer tissues.Results:miR-218-5p was successfully overexpressed in A549 cells(P<0.01).Overexpression of miR-218-5p significantly inhibited the expressions of PDE7A,HK2,PKM2 proteins(all P<0.01),glucose uptake and lactate production(both P<0.01)in A549 cells.Overexpression of PDE7A significantly promoted the expressions of PDE7A,HK2,and PKM2 proteins(all P<0.01),as well as glucose uptake and lactate production(both P<0.01)in A549 cells.miR-218-5p in A549 cells could directly bind to the 3′-UTR of PDE7A mRNA.Database data analysis showed that PDE7A mRNA was highly expressed in lung squamous cell carcinoma tissues(P<0.01).Conclusion:miR-218-5p targets PDE7A to regulate the expression levels of HK2 and PKM2 in A549 cells,which in turn inhibits the glycolytic process.miR-218-5p/PDE7A may be a potential target for clinical diagnosis and treatment of NSCLC.

Objective: To retrospectively analyze the epidemiological characteristics of supernumerary teeth in patients aged 4-18 years old and the influencing factors on the selection of anesthesia methods, and to provide a reference for the selection of anesthesia plans for children and adolescents with supernumerary teeth. Methods: This study is a retrospective study approved by the Institutional Ethics Committee. Based on clinical electronic medical record system and cone beam CT (CBCT) data, a retrospective analysis was conducted on 2 210 patients 4-18 years of age who underwent supernumerary tooth extraction at the School of Stomatology, Fourth Military Medical University from August 2019 to July 2021. Inclusion criteria: ① Age 4-18 years old; and ② The American Society of Anesthesiologists (ASA) classifies anesthesia into grades I-II; and ③ Have complete oral and anesthesia case records and relevant imaging data. Exclusion criteria: ① Incomplete medical records or unclear imaging data; and ② Patients with ASA grade II or above. Patients’ gender and age, the number of supernumerary teeth, arch, location, orientation, eruption status, reason for appointment, anxiety level, degree of cooperation, anesthesia method, and other relevant information were collected and statistically analyzed. Results: A total of 1 865 eligible patients were included, with an average age of (8.9±3.2) years old. There were more male patients (71.37%, 1 331 cases) than female patients (28.63%, 534 cases) (P < 0.001), with a gender ratio of 2.49:1. The majority of supernumerary teeth were single (75.97%, 1 417/1 865), primarily located in the maxilla (97.2%, 1 812/1 865) and the anterior dental region (94.2%, 1 757/1 865), and in a centric position (77.3%, 1 442/1 865). The majority of patients with erupted supernumerary teeth were active in seeking treatment (97.67%, 335/343). Patients with supernumerary teeth located in the maxilla and mandible bones, as well as in the nasal floor, were mainly referred to the department for diagnosis, accounting for 94.38%(1 361/1 442) and 90.00% (72/80) (χ2=1 363.24, P < 0.001), respectively. Regarding anesthesia methods, the largest proportion of patients received nitrous oxide sedation-assisted local anesthesia or nerve block anesthesia, accounting for 38.07% (710/1 865), followed by local anesthesia, accounting for 35.23% (657/1 865). The proportion of patients receiving midazolam intravenous sedation with local anesthesia or nerve block anesthesia and general anesthesia was relatively small, accounting for 20.86% (389/1 865) and 5.84% (109/1 865). Patients 13-18 years of age with supernumerary teeth in the mandibular and posterior regions and without anxiety had the highest proportion of local anesthesia use (P < 0.001). In contrast, patients who had supernumerary teeth located at the base of the nose (50%), severe anxiety (94.12%), and poor cooperation (98.18%) had the highest proportion of general anesthesia use (P < 0.001). There was no significant difference (P = 0.35) in the incidence of intraoperative and postoperative complications after the extraction of supernumerary teeth. However, the proportion of anesthesia-related complications, such as dizziness, coughing, and respiratory depression, occurring in patients who received general anesthesia was higher than local anesthesia, accounting for 3.81% (P = 0.006). Conclusion There is a gender difference in the incidence of supernumerary teeth in patients 4-18 years of age, with a higher prevalence in males. The majority of supernumerary teeth are single and located in the maxillary anterior region, predominantly in a centric position. Patients whose teeth had erupted were more likely to seek medical treatment voluntarily, while patients with supernumerary teeth located in the maxilla and mandible bones, as well as in the nasal floor were more likely to be referred to the department. Patients with high levels of anxiety, poor cooperation, young age, multiple teeth, and high surgical difficulty were more inclined to choose general anesthesia.

Objective : To investigate the efficacy of a holistic approach for postoperative pain management in children receiving dental treatment under general anesthesia in day-surgery operating room. Methods: A total of 120 children, aged 3-7 years, of American Society of Anesthesiologists physical status Ⅰ or Ⅱ, with ≥ 10 treated teeth, receiving comprehensive dental treatment under general anesthesia from January 2020 to August 2020 were enrolled in this trial and randomly allocated into the holistic approach group (group H, n=60) and including preemptive analgesia, instructions to parents for pain management and web-based assessment system (assessment pain by scanning the quick response code 4, 6, and 24 hours postoperatively) and the control group (group C, n=60) only scanning the quick response code. Pain, face, legs, activity, cry and consolability (FLACC) scale was used to assess the level of pain 2 h postoperatively and the parents postoperative pain measure (PPPM) was used to assess the level of pain 4, 6, and 24 h postoperatively in two groups. Results: The FLACC scores of group H 2 h postoperatively were significantly lower than group C (P <0.05). The incidences of significant pain (PPPM scores ≥ 6) 4, 6 and 24 h postoperatively in group H were lower than group C (P <0.05). Altogether, 91.7% of parents in group H and 71.6% in group C assessed the level of pain of children over time. The compliance rate of parents in group H was significantly higher than group C (P <0.05). Conclusion The holistic approach had a positive effect on reducing postoperative pain for children receiving dental treatment under general anesthesia in the day-surgery operating room.

In order to enrich the library of domestic research about new red fluorescent marker in lactic acid bacteria (LAB), we described a new fusion expression system in Lactobacillus plantarum WCFS1 based on the pSIP vector. This system contained red fluorescent protein mCherry as a marker and bile salt hydrolase gene (bsh) as a reporter gene. Moreover, in this study, four different promoters (PsppA, PldhL, P32 and PslpA) were used to regulate the expression of the fusion protein mCherry-BSH, completing the inducible and constitutive expression in lactic acid bacteria. The recombinant protein mCherry-BSH presented activity of red fluorescence and bile salt hydrolase (BSH). The successful construction of the fusion expression system in LAB using a red fluorescent protein mCherry provides favorable conditions for the distribution, intestinal colonization and survival rate of lactic acid bacteria, so as to reveal the function mechanism of its probiotic characteristics; and the system also could lay the foundation for researches on protein expression, cellular localization and properties identification of active protein in lactic acid bacteria.
Lactobacillus plantarum ; Luminescent Proteins ; Probiotics

Objective To investigate the effect of NS-398 combined with docosahexaenoic acid (DHA) on apoptosis of cholangiocarcinoma QBC939 cells and its mechanism.Methods In vitro,cultured cholangiocarcinoma QBC939 cells were exposed to 0,25,50,100,150 and 200 μmol/L NS-398 with 0,15,30,45,60 and 75 μg/ml DHA respectively.The absorbance of the QBC939 cells were measured by CCK8 and its growth inhibition ratio were calculated.Flow cytometry was applied to detect cell apoptosis.The level of β-catenin and c-myc mRNA and protein were measured by real-time PCR,Western blot and enzyme-linked immunoabsordent assay respectively.Results Exposure to NS-398 combined with DHA suppressed the growth of QBC939 cells.When NS-398 was at 100 μmol/L and DHA at 45 pμg/ml,the relative growth inhibition rate of QBC939 cells was 90％ (F =5.85,P ＜ 0.05).NS-398 combined with DHA promoted QBC939 cells apoptosis at the early stage (F =8.16,P ＜ 0.01).Real-time PCR could detect low β-catenin and c-myc expression in QBC939 cells disposed by NS-398 combined with DHA (F =7.61,P ＜0.01),(F =7.92,P ＜0.01).NS-398 combined with DHA decreased β-catenin (F =7.75,P ＜ 0.01),(F=8.17,P＜0.01) and c-myc (F=8.76,P＜0.01),(F=8.12,P＜0.01) protein expression in QBC939 cells.Conclusion NS-398 combined with DHA promoted apoptosis and inhibit proliferation of cholangiocarcinoma cells QBC939 in vitro possibly through downregulated mRNA and protein expression of β-catenin and c-myc.

Objective:To investigate the incidence and the influencing factors of school bullying among middle school students in Xi'an,thus to provide evidence for strengthening the prevention of school violence.Methods:Using Chinese version of School Bullying Experience Questionnaire (C-SBEQ),a survey was conducted among 995 middle school students selected from 2 middle schools in Xi'an.The data was analyzed using SPSS 22.0.Results:The incidence of school bullying in one year was 42.0％,approximately 36.8％ of the students suffered from verbal bullying,while 5.8％ of the students suffered from physical bullying.As for perpetrating,19.0％ and 5.4％ of the students were the perpetrators of verbal bullying and physical bullying,respectively.The influencing factors of school bullying included gender,mother's education level,study type,playing video games,smoking and drinking (P ＜0.05).Conclusion:School bullying among middle school students in Xi'an was at a high level.The occurrence of school bullying was related to personal behavior,family,school and other factors.Thus,targeted prevention measures should be taken to prevent school bullying.

Objective To knockout Rag2 and IL2rg genes and construct severe combined immunodeficiency mice based on CRISPR/Cas9 technology. Method Design and synthesis of 25 bp sgRNA were made according to the Rag2 and IL2rg sequences in Genbank. After annealing, sgRNA was cloned into pX330 vector. Recombination plasmid Rag2?sgRNA, IL2rg?sgRN and Cas9 were then transcribed into RNA, these RNA were microinjected into zygotes and the zygotes were transplanted into recipient ICR mice. F0 founders were born and mutated F0 founders mated with wild type mice to obtain F1 generation heterozygous mice. Mutated F1 mice were crossed and got F2 generation homozygous mice. Genotype and phenotype of the knockout mice were identified by sequencing, flow cytometry and xenograft model. Results Rag2?sgRNA and IL2rg?sgRNA recombination plasmids were constructed and transcribed into RNA. After microinjection and mat? ing, F0 founders were born and F2 homozygous mice were obtained. The results of sequencing showed that there were two types of genotype in IL2rg gene, 10 bp or 11 bp deletion;however, there was only one genotype in Rag2 gene, which was 8 bp deletion. Compared with wild?type BALB/c mice, the number of CD3 +, B220 + and NKp46 + cells in peripheral blood of the knockout mice was reduced significantly. After inoculation of human breast cancer cell line SKBR?2HL cells, tumor size in the xenograft mouse model was increased gradually along with time extension. Conclusion CRISPR/Cas9 is an efficient way to mutate Rag2 and IL2rg gene in mice in vivo, leading to aberrant T cells, B cells and NK cells.

Objective To construct miRNA-29b1 gene knockout mice based on CRISPR/Cas9 technology. Methods To design and synthesize sgRNA according to the miRNA-29b1 sequence in Genbank .sgRNA and Cas9 were transcribed to RNA in vitro, these RNA were then microinjected into zygotes of C 57BL/6 mice.After mouse birth, the genome DNA was extracted and sequenced to identify its genotype; meanwhile , real-time PCR was used to assay the expression of miRNA-29b1 in the heart, liver, spleen, lung and kidney of mutated mice .Result A 20 bp sgRNA targeted on miRNA-29b1 was synthesized and transcribed to RNA with Cas 9.After microinjection, miRNA-29b1 gene-mutated mice were obtained.The sequencing results showed that there were two types of genotype for the mutated mice , one was 10 bp deletion, and another was 23 bp deletion accompanied with a 3 bp insertion.Compared with the wild-type mice, the expression of miRNA-29b1 in the heart, liver, spleen, lung and kidney was reduced significantly .Conclusions miRNA-29b1 gene knockout mice are constructed successfully by using CRISPR /Cas9 technology.

Purpose] The article analyzed the rules of diagnosis and treatment of “menstruation stopped when not old yet”in the Fuqingzhunvke Section · Menstruation Chapter, which provides a new thought of diagnosing and treating for the treatment of premature ovarian failure. [Methods] Because of the characteristics of the provisions in the Fuqingzhunvke is consistent with the clinical symptoms of premature ovarian failure in the modern medicine. The article analyzed the cause, pathogenesis, principle-method-recipe-medicine and treatment to explore the treatment of premature ovarian failure. [Result] The characteristics of the provisions in the Fu Qingzhu Nvke are consistent with the clinical symptoms of premature ovarian failure in the modern medicine. Fushi thought that premature ovarian failure derived from heart, liver, spleen and kidney, treatment should reinforce kidney water;puts forward some new ideas for qi depression, nourishing liver to resolve depression method;pay attention to tonifying spleen, nourishing heart and tranquilizing mind; reminding relieving depression lies in strengthening the body resistance. [Conclusion] Through analyzing the etiology, pathogenesis, treatment of premature ovarian failure, to clarify the etiology, pathogenesis, treatment of premature ovarian failure. The study of classics of traditional Chinese medicine provides a new method for the treatment of premature ovarian failure. Making the past serve the present, the clinical curative effect will be improved.

Objective] Investigate the relationship between polycystic ovary syndrome and phlegm wet, lay a theoretical foundation for from the wet treatment. [Method] According to the diagnostic standard of PCOS, try from the signs and symptoms, laboratory indexes, imaging manifestations of three aspects, analyzes and demonstrates the relationship between the disease with phlegm wet. From curing phlegm wet as the breakthrough point, dialectical analysis and tailored medication,explore the treatment effect and advantage. [Result] PCOS shown by a series of symptoms and signs are associated with phlegm wet blocking uterus; Insulin resistance, lipid metabolic disorder, leptin, and tumor necrosis factor pathways are closely associated with phlegm wet; Polycystic ovary is visual performance of phlegmy wet block uterus.Arguments from phlegmy wet, and guiding drugs, combined with the spirit, exercise, diet therapy have a significant clinical curative effect .[Conclusion] In this paper, starting from the theory of phlegm wet treatment of the disease,curative effect is distinct and has high practical value. Also it demonstrates the scientific and particularity of Chinese medicine on disease. It will provide a new train of thought for the gynecologic diseases and the other diseases in the future.