Vitrification cryopreservation technology converts the liquid water in biological samples into glassy solids through ultra-rapid cooling. In this review, the core research of vitrification cryopreservation technology was mainly reviewed, such as the optimization of cryoprotectant formulations, the improvements to temperature control methods, the innovative carrier designs and the improvements to rewarming technology. The application status and issues in the fields of organ preservation, tumor research, reproductive medicine and rare cell preservation were also summarized. By combining the technical principles with glassy state detection methods, a theoretical basis was provided for optimizing the technical system and expanding application scenarios, thereby promoting its large-scale application in clinical and biological sample libraries.

Objective To establish the framework of a prediagnostic risk assessment system for outpatient dental care and to provide references for ensuring patient safety and improving the quality of medical services.Methods A total of 15 medical workers in a tertiary-care stomatology hospital in Sichuan Province were selected for qualitative interviews between October 2019 and December 2019.On the basis of the results of literature analysis and the interviews,an expert consultation form for prediagnostic risk assessment system for outpatient dental care was developed.Then,consultation of 10 experts in the field of oral health care and nursing was conducted between June 2020 and December 2020.Eventually,the framework of prediagnostic risk assessment system for outpatient dental care was formulated.Results Four themes emerged from the qualitative interviews.Firstly,prediagnostic risks of dental outpatients involved mainly syncope,cardiovascular emergencies,and other emergency medical risks.Secondly,medical risks came from three sources,patients,healthcare professionals,and the environment.Thirdly,medical professionals of outpatient dental care had limited awareness of the prediagnostic medical risks of patient.Fourthly,medical professionals of outpatient dental care showed inadequate response to and management of the prediagnostic medical risks of patient.The expert consultation helped finalize the Dental Outpatient Prediagnostic Risk Assessment Questionnaire,which included 3 primary indicators(namely,general information,medical history[including history of allergy],and medication assessment),12 secondary indicators(including patient demographics,patients'status upon admission,oral hygiene habits and special lifestyle habits,sensory disorders,special periods for female patients[ie,menstruation,pregnancy,and breastfeeding],allergy history[history of oral treatment-related allergies],past/present medical history,types of medication,route of medication administration,duration of medication administration,accuracy of medication administration,and adverse drug reactions),and 39 tertiary indicators.The effective recall rate of the expert consultation form was 100%,expert positivity was 100%,and the authority coefficient was 0.83.Kendall's harmony coefficient ranged from 0.808 to 0.839,which was statistically significant(P<0.001).Conclusion The framework of prediagnosis risk assessment system for outpatient dental care is found to be scientific and specific,but its applicability still needs further validation in clinical practice.

Extracellular vesicle (EV) is widely distributed and fully involved in the regulation of various aspects of cellular physiology. As active carriers of molecular biomarkers and mediators of intercellular communication, EV is involved in the occurrence, development, metastasis, drug resistance and other links of tumors. The EV, which can be obtained from a variety of body fluids and are highly homologous to tissues, plays an important role in the maintenance of cell homeostasis and the exchange of substances and information between cells, thus becoming an ideal biomarker for tumor diagnosis and treatment. In the emerging liquid biopsy application, EV DNA can be used as a molecular marker for the diagnosis and prognosis evaluation of many kinds of tumors, and it is the most potential molecular marker of liquid biopsy in the field of tumor diagnosis and treatment.

Objective:To analyze the clinical characteristics of acute ischemic stroke (AIS) patients with negative cerebral CT perfusion (CTP) and influencing factors for their prognoses.Methods:A retrospective analysis was performed; 448 patients with AIS admitted to Department of Neurology, Sixth Affiliated Hospital of Nantong University from January 2020 to June 2021 were enrolled. CTP images of these patients were processed by RAPID software, and they were divided into CTP-negative group and CTP-positive group according to cerebral infarction core and ischemic penumbra volumes. The clinical data were compared between patients from CTP-negative group and CTP-positive group and between patients from CTP-negative and CTP-positive subgroups accepted thrombolytic therapy. According to the prognoses 3 months after discharge, CTP negative patients were divided into poor prognosis group and good prognosis group. Independent influencing factors for poor prognosis in negative CTP patients were analyzed by univariate and multivariate Logistic regressions.Results:(1) In these 448 patients, 154 (34.4%) were with negative CTP and 294 (65.6%) were with positive CTP; compared with the CTP-positive group, the CTP-negative group had significantly younger age, significantly higher percentage of patients with diabetes, significantly lower percentage of patients with atrial fibrillation, statistically higher baseline systolic blood pressure, and significantly lower baseline National Institutes of Health Stroke Scale (NIHSS) scores, early neurological deterioration (END) incidence, modified Rankin scale (mRS) scores 3 months after discharge, and proportion of patients with poor prognosis ( P<0.05); significant differences in distributions of responsible circulations for the lesions and etiological classification (TOAST) were noted between the 2 groups ( P<0.05). Of the 448 patients, 270 received thrombolytic therapy, including 101 CTP-negative patients and 169 CTP-positive patients; compared with the CTP-positive subgroup, the CTP-negative subgroup had significantly younger age, significantly lower percentage of patients with atrial fibrillation, statistically higher baseline systolic blood pressure, and significantly lower baseline NIHSS scores, END incidence, mRS scores 3 months after discharge, and proportion of patients with poor prognosis ( P<0.05). (2) Of the 154 CTP negative patients, 31 had poor prognosis and 123 had good prognosis. Univariate Logistic regression analysis showed that baseline blood glucose, fasting blood glucose, glycosylated hemoglobin (HbA1C), baseline NIHSS scores and fibrinogen were the influencing factors for prognoses of CTP negative patients, with significant differences ( P<0.05). Multivariate Logistic regression analysis found that NIHSS ( OR=0.827, 95% CI: 0.743-0.920, P<0.001) and HbA1 C ( OR=0.763, 95% CI: 0.609-0.956, P=0.019) were independent influencing factors for poor prognosis of CTP-negative patients. Conclusion:AIS patients with negative CTP have milder neurological impairment, better prognosis, and higher safety of receiving intravenous thrombolysis than those with positive CTP; AIS patients with negative CTP enjoying high baseline NIHSS scores and HBA1c have poor prognosis.

To establish a method for isolation, culture and identification of adipose-derived mesenchymal stem cells (ASCs) from the inbreed line miniature pig of Wuzhishan (ILMW).
 Methods: A total of 100 g adipose tissues were obtained from subcutaneous tissues of neck in six-month old healthy ILMW (3 samples, male). ASCs from ILMW (ILMW-ASCs) were isolated from adipose tissues through 0.1% collagenase digestion. The cells at the 3rd, 5th, 8th, 13th passages were collected. Cell morphology, size, phenotype, cell cycle, and apoptosis were monitored. Cell differentiation was induced and cell proliferation curve was drawn.
 Results: The ILMW-ASCs, fibroblast-like or whirlpool-like, began the adherence at 36 h and entered a logarithmic phase in the 5th day. Eighty percent of them were fused in the 7th day. The average diameter and volume of ILMW-ASCs were (17.00±0.54) µm and (2.58±0.24)×10-9 L, respectively. The expressions of CD29, CD44 and CD90 were positive, and there was no significant difference between the different passages (all P>0.05). The expressions of CD45, CD8a and HLA-DR were increased with the increase in passages after the 3th passage (all P<0.05). The adipogenic induction of ILMW-ASCs was observed by positive oil red O staining, and the osteogenic induction of ILMW-ASCs was determined by positive alizarin red staining. Apoptosis and senescence occurred in the 13 passage of ILMW-ASCs, and the proportion of S phase of cell cycle was lower than that in lower passages (all P<0.05). 
 Conclusion: ILMW-ASCs are one of the best choice for porcine ASCs, which might provide a source of candidate stem cells for therapy of large animal disease models and tissue or organ repairment.
Adipose Tissue ; Animals ; Cell Differentiation ; Cells, Cultured ; Male ; Mesenchymal Stem Cells ; Swine ; Swine, Miniature

Preclinical evaluation is related to the clinical safety of radiopharmaceuticals.There are different research foci on preclinical evaluation of different radiopharmaceuticals.This article summarizes the Food and Drug Administration (FDA) preclinical evaluation guidelines of diagnostic and therapeutic radiopharmaceuticals,in order to provide reference for domestic research and preclinical evaluation of radiopharmaceuticals.

OBJECTIVE: To identify a rare large off ladder (OL) allele at the Penta E locus. METHODS: Chelex-100 was used to extract DNAs from the blood samples. The PCR fragments were purified, extracted, and subjected to TA cloning and sequencing. RESULTS: An OL allele was identified by the PowerPlex™ 21 at the Penta E locus, which was postulated as allele 26 based on assigned size. The OL allele was verified as a novel fragment containing 26 full AAAGA repeats. CONCLUSION OL alleles and microvariants should be verified by direct sequencing. Typing of OL alleles has significance for both daily work and forensic genetics.
Alleles ; Polymerase Chain Reaction ; Rubiaceae

Radiopharmaceuticals have been widely used in the diagnosis, treatment and monitoring of diseases, and they play an important role in new drug development. Food and Drug Administration(FDA) has rich experience in the administration of radiopharmaceuticals. This article mainly interprets the regulato-ry policy of FDA for radiopharmaceuticals from the aspects of definition, regulations and registration, trying to provide reference for domestic research of radiopharmaceuticals.

DEAD-box family protein is a kind of ATP dependent RNA helicase,which plays a critical role in RNA metabolism.The DEAD-box family proteins can affect cell proliferation,differentiation,and apoptosis through regulating the expression of oncogenes,tumor suppressor genes and tumor related signaling pathways.It plays the role in promoting or suppressing cancer.

Objective:To provide the basic data for the further revision of mycoplasma test method described in Chinese Pharma-copoeia and some references for the operation standardization of drug mycoplasma test. Methods:Two incubation conditions,namely aerobic conditions and microaerophilic conditions,were compared with respect to the growth status of mycoplasma in liquid and solid media. Results:The growth of mycoplasma was obvious difference between the two incubation conditions,and the microaerophilic con-ditions were better than the aerobic conditions. Conclusion:The microaerophilic conditions can be used in the incubation of mycoplas-ma test,which should be defined and standardized in the future Chinese Pharmacopoeia.