Objective:To establish a method for the rapid detection of varicella-zoster virus (VZV) by recombinase-aid amplification (RAA) combined with Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system.Methods:Clinical samples of suspected herpes zoster in Shandong province and Shanghai from 2023 to 2024 were collected, nucleic acids of positive samples were extracted, RAA-specific primers and crRNA (CRISPR RNA, crRNA) were designed for the conserved region of VZV, and the fluorescence intensity generated by Cas12a non-specific cleavage of single-stranded fluorescent probes was used to screen highly sensitive crRNAs and optimize the concentrations of crRNA, Cas12a and ssDNA probes. The sensitivity and reproducibility of the RAA-CRISPR/Cas12a detection method were evaluated by using synthesized plasmids and clinical samples, and the specificity of the method was evaluated by using other viral nucleic acids. The method was used to detect clinical samples by using the method and quantitative real-time PCR (qPCR) method, and the detection rate and consistency of the two method were compared.Results:The highly sensitive crRNA-4 was screened from the four crRNAs designed, and a VZV detection method for RAA-CRISPR/CAS12a based on fluorescence intensity measurement was established, which could be detected at 37℃ in 45 min, and the sensitivity of the detection could reach 10 copies/μL, a minimum clinical sample with a Ct value of 38.980 can be detected. It has high specificity and no cross-reactivity with Adenovirus 7, Herpes simplex virus type I, Herpes simplex virus type II, Coxsackieviruses A16, Cytomegalovirus, Epstein-Barr virus, Measles virus, Mumps virus, Enterovirus 71, Japanese encephalitis virus genotype 5. It has good stability, and can be successfully detected in low, medium and high concentrations of viral positive plasmids with good consistency. The detection rate of the clinically positive samples was 100%, which was completely consistent with the qPCR test result.Conclusions:RAA isothermal amplification technology combined with CRISPR-CAS12a technology was used to establish an accurate method for the detection of VZV virus, which was highly sensitive, specific, and had low requirements for experimental conditions, and could be completed within 45 min, which could provide strong technical support for the early detection of VZV.

Objective To develop an ultra-high performance liquid chromatography-mass spectrometry method(UPLC-MS/MS)for the determination of nirmatrelvir concentration in mouse plasma.Methods The ACQUITY UPLC system was used in tandem with an API 4000 triple quadrupole mass spectrometer.The analytical column was Waters BEH C18(2.1 mm×5.0 mm,1.7 μm)column,and the mobile phases consisted of water(containing 0.1％formic acid)and methanol(containing 0.1％formic acid)under gradient elution at the flow rate of 0.4 mL·min-1.The column temperature was set at 40 ℃,and the injection volume was 5 μL.Electrospray ionization was used as ion source,and positive multiple reaction monitoring mode was adopted to quantitatively analyze the ionization pairs m/z 500.3→110.3(nirmatrelvir)and m/z 237.3→193.3(carbamazepine).Carbamazepine was employed as an internal standard.Results The linear range of nirmatrelvir was from 10 ng·mL-1 to 2 560 ng·mL-1.For the quality control nirmatrelvir samples,the accuracies of intra-and inter-batch were less than±15％,and the precisions of intra-and inter-batch were lower than 15％.Nirmatrelvir in plasma was stable at room temperature for 24 h and remained stable after three freeze-thaw cycles.The extracted nirmatrelvir solution could be stored at 4℃ for 3 d without any visible change.Conclusion The method was characterized by good specificity,high sensitivity,and appropriate linear range.The methodological validation was in accordance with the 2020 edition of the Chinese Pharmacopoeia and could be applied to the quantitative detection of nirmatrelvir in plasma.

Dental stem cells(DSCs),a distinct subset of mesenchymal stem cells(MSCs),are isola-ted from dental tissues,such as dental pulp,exfoliated deciduous teeth,periodontal ligament,and apical papilla.They have emerged as a promising source of stem cell therapy for tissue regeneration and autoim-mune disorders.The main types of DSCs include dental pulp stem cells(DPSCs),stem cells from hu-man exfoliated deciduous teeth(SHED),periodontal ligament stem cells(PDLSCs),and stem cells from apical papilla(SC AP).Each type exhibits distinct advantages:easy access via minimally invasive procedures,multi-lineage differentiation potential,and excellent ethical acceptability.DSCs have demon-strated outstanding clinical efficacy in oral and maxillofacial regeneration,and their long-term safety has been verified.In oral tissue regeneration,DSCs are highly effective in oral tissue regeneration for critical applications such as the restoration of dental pulp vitality and periodontal tissue repair.A defining advan-tage of DSCs lies in their ability to integrate with host tissues and promote physiological regeneration,which render them a better option for oral tissue regenerative therapies.Beyond oral applications,DSCs also exhibit promising potential in the treatment of systemic diseases,including type Ⅱ diabetes and au-toimmune diseases due to their immunomodulatory effects.Moreover,extracellular vesicles(EVs)de-rived from DSCs act as critical mediators for DSCs' paracrine functions.Possessing regulatory properties similar to their parental cells,EVs are extensively utilized in research targeting tissue repair,immuno-modulation,and regenerative therapy—offering a"cell-free"strategy to mitigate the limitations associated with cell-based therapies.Despite these advancements,standardizing large-scale manufacturing,maintai-ning strict quality control,and clarifying the molecular mechanisms underlying the interaction of DSCs and their EVs with recipient tissues remain major obstacles to the clinical translation of these treatments into broad clinical use.Addressing these barriers will be critical to enhancing their clinical applicability and therapeutic efficacy.In conclusion,DSCs and their EVs represent a transformative approach in re-generative medicine,and increasing clinical evidence supports their application in oral and systemic diseases.Continuous innovation remains essential to unlocking the widespread clinical potential of DSCs.

Objective To develop an ultra-high performance liquid chromatography-mass spectrometry method(UPLC-MS/MS)for the determination of nirmatrelvir concentration in mouse plasma.Methods The ACQUITY UPLC system was used in tandem with an API 4000 triple quadrupole mass spectrometer.The analytical column was Waters BEH C18(2.1 mm×5.0 mm,1.7 μm)column,and the mobile phases consisted of water(containing 0.1％formic acid)and methanol(containing 0.1％formic acid)under gradient elution at the flow rate of 0.4 mL·min-1.The column temperature was set at 40 ℃,and the injection volume was 5 μL.Electrospray ionization was used as ion source,and positive multiple reaction monitoring mode was adopted to quantitatively analyze the ionization pairs m/z 500.3→110.3(nirmatrelvir)and m/z 237.3→193.3(carbamazepine).Carbamazepine was employed as an internal standard.Results The linear range of nirmatrelvir was from 10 ng·mL-1 to 2 560 ng·mL-1.For the quality control nirmatrelvir samples,the accuracies of intra-and inter-batch were less than±15％,and the precisions of intra-and inter-batch were lower than 15％.Nirmatrelvir in plasma was stable at room temperature for 24 h and remained stable after three freeze-thaw cycles.The extracted nirmatrelvir solution could be stored at 4℃ for 3 d without any visible change.Conclusion The method was characterized by good specificity,high sensitivity,and appropriate linear range.The methodological validation was in accordance with the 2020 edition of the Chinese Pharmacopoeia and could be applied to the quantitative detection of nirmatrelvir in plasma.

Objective:To establish a method for the rapid detection of varicella-zoster virus (VZV) by recombinase-aid amplification (RAA) combined with Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a system.Methods:Clinical samples of suspected herpes zoster in Shandong province and Shanghai from 2023 to 2024 were collected, nucleic acids of positive samples were extracted, RAA-specific primers and crRNA (CRISPR RNA, crRNA) were designed for the conserved region of VZV, and the fluorescence intensity generated by Cas12a non-specific cleavage of single-stranded fluorescent probes was used to screen highly sensitive crRNAs and optimize the concentrations of crRNA, Cas12a and ssDNA probes. The sensitivity and reproducibility of the RAA-CRISPR/Cas12a detection method were evaluated by using synthesized plasmids and clinical samples, and the specificity of the method was evaluated by using other viral nucleic acids. The method was used to detect clinical samples by using the method and quantitative real-time PCR (qPCR) method, and the detection rate and consistency of the two method were compared.Results:The highly sensitive crRNA-4 was screened from the four crRNAs designed, and a VZV detection method for RAA-CRISPR/CAS12a based on fluorescence intensity measurement was established, which could be detected at 37℃ in 45 min, and the sensitivity of the detection could reach 10 copies/μL, a minimum clinical sample with a Ct value of 38.980 can be detected. It has high specificity and no cross-reactivity with Adenovirus 7, Herpes simplex virus type I, Herpes simplex virus type II, Coxsackieviruses A16, Cytomegalovirus, Epstein-Barr virus, Measles virus, Mumps virus, Enterovirus 71, Japanese encephalitis virus genotype 5. It has good stability, and can be successfully detected in low, medium and high concentrations of viral positive plasmids with good consistency. The detection rate of the clinically positive samples was 100%, which was completely consistent with the qPCR test result.Conclusions:RAA isothermal amplification technology combined with CRISPR-CAS12a technology was used to establish an accurate method for the detection of VZV virus, which was highly sensitive, specific, and had low requirements for experimental conditions, and could be completed within 45 min, which could provide strong technical support for the early detection of VZV.

The orofacial system, an intricate assembly of diverse tissues that underpin the unique biologic and morphological identity of an individual, presents a formidable challenge in the realm of tissue regeneration within oral and maxillofacial surgery. This review conducts a retrospective appraisal of advancements in the field of orofacial tissue regeneration, elucidating the current research landscape while critically addressing the persisting challenges. It underscores the pivotal roles of orofacial mesenchymal stem cells in orchestrating regenerative processes, offering an insightful outlook on future developments. The objective is to demarcate innovative therapeutic avenues and clinical implications by fostering a comprehensive understanding of this domain among dental practitioners. As such, it aspires to serve as a valuable reference for prospective investigations and to elevate the knowledge base pertaining to orofacial tissue regeneration.

ABSTRACT: Meningiomas are the most common primary intracranial neoplasm with diverse pathological types and complicated clinical manifestations. The fifth edition of the WHO Classification of Tumors of the Central Nervous System (WHO CNS5), published in 2021, introduces major changes that advance the role of molecular diagnostics in meningiomas. To follow the revision of WHO CNS5, this expert consensus statement was formed jointly by the Group of Neuro-Oncology, Society of Neurosurgery, Chinese Medical Association together with neuropathologists and evidence-based experts. The consensus provides reference points to integrate key biomarkers into stratification and clinical decision making for meningioma patients. REGISTRATION Practice guideline REgistration for transPAREncy (PREPARE), IPGRP-2022CN234.
Humans ; Meningioma/pathology* ; Consensus ; Neurosurgical Procedures ; Meningeal Neoplasms/pathology*

OBJECTIVE: To establish a mouse model bearing orthotopic temozolomide (TMZ)-resistant glioma that mimics the development of drug resistance in gliomas METHODS: Seventy-eight adult C57BL/6 mice were randomly divided into 6 groups ( RESULTS: The mouse models bearing TMZresistant glioma was successfully established. The cells from the high-dose induced group showed a significantly higher colony-forming rate than those from the high-dose control group ( CONCLUSIONS Progressive increase of TMZ doses in mice bearing orthotopic gliomas can effectively induce TMZ resistance of the gliomas.
Animals ; Antineoplastic Agents, Alkylating/pharmacology* ; Brain Neoplasms/drug therapy* ; Cell Line, Tumor ; Disease Models, Animal ; Drug Resistance, Neoplasm ; Glioma/drug therapy* ; Mice ; Mice, Inbred C57BL ; Temozolomide/therapeutic use*

Objective:To evaluate the effect of goal-oriented online and offline mixed teaching method on the trainees of burn operating room.Methods:From June 2019 to June 2020, 42 trainees of burn operating room in our hospital were selected for the randomized parallel trial, and they were randomly divided into two groups, routine group and research group. The routine group adopted the conventional online and offline mixed teaching method, while the research group adopted the goal-oriented online and offline mixed teaching method. The internship time of both group lasted for 1 month. Results and excellent rates, self-confidence and burn surgery skills evaluation before and after the internship, and satisfaction with the internship mode were compared between the two groups. SPSS 19.0 was used for t test and chi-square test. Results:The results and excellent and good rates of theoretical examination and practical examination in the research group were higher than those of the routine group. The scores of self-confidence, choice of operation mode, innovation and optimization of operation, control of operation complications and treatment of intraoperative emergencies in the two groups after internship were higher than those before internship, and the above scores of research group were higher than those of the routine group after internship. The satisfaction scores of the students on enhancing self-confidence, and improving operational ability, learning initiative and learning efficiency in the research group were significantly higher than those in the routine group ( P < 0.05). Conclusion:Goal-oriented online and offline mixed teaching method on trainees in burn operating room can not only improve the examination results, enhance their confidence and burn surgery skills, but also achieve their satisfaction.

Objective:In order to establish a painless delivery detection system in plateau area, promote the concept of painless delivery, reduce the cesarean section rate, the clinical study of painless childbirth in Tibet was carried out.Methods:150 primiparas in Lhasa People's Hospital from January 2018 to Juen 2019 were prospectively collected, which were randomly divided into sufentanil and ropivacaine groups (group A, n=50) and fentanyl and ropivacaine (group B, n=50) and the control group without analgesia (group C, n=50); The general conditions of the patients before and after delivery were collected. The visual analogue scale (VAS) score was used to observe and record the delivery process and results. The Apgar score of newborns was performed, and the umbilical artery blood was taken for blood gas analysis. Results:The first and second stage of labor in group A and B were shorter than those in group C ( P<0.05); there was no difference in the third stage of labor and postpartum bleeding between the three groups ( P>0.05). The VAS scores of groups A and B were lower than that of group C ( P<0.05), while there was no difference between group A and group B ( P>0.05). There was no difference in pH value of umbilical artery blood between the three groups ( P>0.05). The umbilical artery blood PO 2 of group A and group B was higher than those of group C ( P<0.05), while there was no difference between group A and group B ( P>0.05). The umbilical artery blood PCO 2 of group A and group B was lower than that of group C( P<0.05), while there was no difference between group A and group B ( P>0.05). The cesarean section rate in group C was higher than that in groups A and B ( P<0.05). There was no difference in fetal heart rate, fetal distress, and neonatal asphyxia among the three groups ( P>0.05). There was significant difference in the overall incidence of adverse conditions in the three groups ( P<0.05). The Apgar scores of the newborns in groups A and B were higher than those in the C group at 1 min postpartum ( P<0.05), while there were no difference in the Apgar scores among the three groups at 5 and 10 min postpartum ( P>0.05). Conclusions:Painless delivery in plateau area can reduce the pain of delivery and the rate of cesarean section, and has high safety and reduce the adverse effects on mothers and infants, which is worthy of clinical promotion.