206,977 newborn screening results reveal the ethnic differences in the spectrum of inborn errors of metabolism in Huaihua, China.

Background: Inborn errors of metabolism (IEMs) are rare diseases caused by inherited defects in various biochemical pathways that strongly correlate with early neonatal mortality and stunting. Currently, no studies have reported on the incidence of IEMs of multi-ethnic groups in Huaihua, China. Methods: A total of 206,977 neonates with self-reported ethnicity who underwent IEM screening at Huaihua from 2015 to 2021 were selected for observation. Among them, 69 suspected IEM-positive neonates were referred for urine gas chromatography-mass spectrometry analysis, biochemical detection, next-generation sequencing, and Sanger sequencing. Results: Sixty-nine newborns were diagnosed with IEMs, with an overall incidence of 1:3,000. The two most common disorders were 2-methylbutyryl glycinuria (1:7,137) and phenylalanine hydroxylase deficiency (1:22,997). Moreover, the incidence of IEMs in the minority ethnic group (Miao, Dong, Tujia and Yao) (1:1,852) was markedly higher than in the Han ethnic group (1:4,741). Some ethnic features variants were identified; NM_001609.4:c.1165A>G in the ACADSB gene for Miao and Dong ethnic groups, NM_014251.2:c.852_855del in the SLC25A13 gene for Miao ethnic groups. Conclusion: This study revealed the IEM incidence within the minority ethnic groups is markedly higher than among the Han nationality and the gene variant spectrum is dramatically different in Huaihua, China. Hence, It serves as a theoretical reference for the screening and diagnosing of neonatal IEMs of multi-ethnic groups in the Huaihua area, and across China.

Harmonious passion and academic achievement in higher education: The mediating influence of exploratory and exploitative learning strategies.

Harmonious passion refers to engaging in an activity out of personal volition and sense of meaning, which motivates individuals to devote substantial time and energy while internalizing the activity as part of their identity, and it underpins the promotion of sustainable behaviors and mindsets. The present study aimed to investigate the relationship between harmonious passion and academic achievement among university students in higher education, as well as the mediating role of exploratory and exploitative learning behaviors in this association. Exploratory learning involves seeking new knowledge and skills, whereas exploitative learning involves refining and applying existing knowledge and skills. Data were collected from 528 university students across multiple institutions in China. Using structural equation modeling and multiple regression analysis, results revealed that harmonious passion had a significant positive effect on academic achievement. Furthermore, exploratory and exploitative learning played a chain-mediated role in the relationship. These findings provide insights into how harmonious passion could promote students' academic success in the context of innovation-driven and socially sustainable development, via the facilitation of sustainable learning strategies. Therefore, this study has broader implications for personal and societal sustainability, by emphasizing the role of harmonious passion and sustainable learning strategies in enhancing academic achievement and, ultimately, contributing to a more sustainable society.

Genetic diversity drives extreme responses to traumatic brain injury and post-traumatic epilepsy.

Traumatic brain injury (TBI) is a complex and heterogeneous condition that can cause wide-spectral neurological sequelae such as behavioral deficits, sleep abnormalities, and post-traumatic epilepsy (PTE). However, understanding the interaction of TBI phenome is challenging because few animal models can recapitulate the heterogeneity of TBI outcomes. We leveraged the genetically diverse recombinant inbred Collaborative Cross (CC) mice panel and systematically characterized TBI-related outcomes in males from 12 strains of CC and the reference C57BL/6J mice. We identified unprecedented extreme responses in multiple clinically relevant traits across CC strains, including weight change, mortality, locomotor activity, cognition, and sleep. Notably, we identified CC031 mouse strain as the first rodent model of PTE that exhibit frequent and progressive post-traumatic seizures after moderate TBI induced by lateral fluid percussion. Multivariate analysis pinpointed novel biological interactions and three principal components across TBI-related modalities. Estimate of the proportion of TBI phenotypic variability attributable to strain revealed large range of heritability, including >70% heritability of open arm entry time of elevated plus maze. Our work provides novel resources and models that can facilitate genetic mapping and the understanding of the pathobiology of TBI and PTE.

Ceftazidime-avibactam in the treatment of bacteremia due to carbapenem-resistant gram-negative bacteria in hematological patients: Experience in a single center.

INTRODUCTION: Limited experience exists with ceftazidime-avibactam (CAZ-AVI) in treating bacteremia caused by carbapenem-resistant Enterobacterales (CRE) and Pseudomonas aeruginosa (CRPA) in hematological patients. METHODS: We performed a single-center, retrospective, observational study including patients who received CAZ-AVI for bacteremia due to CRE or CRPA between 2018 and 2022. The primary outcome was 30-day survival. We conducted a multivariable analysis to identify predictors of survival. RESULTS: 56 patients were included and 57 (41 CRE and 16 CRPA) strains were isolated. 35 strains produced carbapenemase, including 25 metallo-beta-lactamase (MBL) and 10 serine-beta-lactamase. 48 patients (85.7 %) received combination therapy. All patients with MBL-CRE bacteremia (n = 24) received combination therapy with aztreonam (AZT). The susceptibility rates to CAZ-AVI were only 26.8 % (11/41) in CRE and 80.0 % (8/10) in CRPA. The 30-day survival rates were 85.0 % (34/40) in the CRE group and 81.3 % (13/16) in the CRPA group. In patients with MBL-CRE bacteremia, the 30-day survival was as high as 91.7 % (22/24) due to combination with AZT. Ceftazidime did not influence the activity of aztreonam-avibactam against MBL-CRE in-vitro. Multivariable cox analysis revealed neutropenia >14 days (P = 0.002, HR: 34.483, 95%CI: 3.846-333.333) and a higher Pitt bacteremia score (P = 0.005, HR: 2.074, 95%CI: 1.253-3.436) were risk factors for 30-day survival. CONCLUSIONS: CAZ-AVI is highly effective in treating bacteremia due to CRPA and serine-beta-lactamase CRE. The combination of avibactam with AZT is highly effective in treating bacteremia due to AZT-resistant MBL producers.

The burden of fracture in China from 1990 to 2019.

PURPOSE: Osteoporosis is one of the most common clinical problems among the elderly population. China is one of the countries most threatened by osteoporosis and fragility fracture, because of its large population and aging population trends during recent decades. We aimed to estimate the disease burden of fracture from 1990 to 2019 in China. METHODS: We performed a secondary analysis of fractures using detailed information for China from the Global Burden of Disease Study 2019. Fracture incidence and prevalence, rate of years lost to disability from fractures, and term secular trends in China from 1990 to 2019 were compared by sex, age, cause, and nature of fracture. RESULTS: The numbers for incidence and prevalence of fracture and years lived with disability (YLDs) from fractures in China increased from 12.54 million, 28.35 million, and 1.71 million in 1990 to 21.27 million, 67.85 million, and 3.79 million in 2019, respectively, increases of 70%, 139%, and 122%, respectively. In 2019, falls was the leading cause of fractures, with an age-standardized incidence rate (ASIR) of 762 per 100 000 (95% uncertainty interval [UI] 629-906), an age-standardized prevalence rate (ASPR) of 1863 per 100 000 (95% UI 1663-2094), and an age-standardized YLD rate (ASYR) of 103 per 100 000 (95% UI 69-147). Fall-associated deaths and disability-adjusted life-years (DALYs) from low bone mineral density increased greatly during the most recent three decades. Fracture of patella, tibia or fibula, and ankle were the most frequent fracture types, with an ASYR of 116 per 100 000 (95% UI 75-169). Hip fracture had more incident cases in adults ≥ 60 years old, and was more frequent for females. CONCLUSIONS: The burden from fractures has increased significantly since 1990 in China. Falls and road injuries are the main causes of the increase. The fall-associated health burden from osteoporosis needs to be prioritized, with longer-term commitment to its reduction required.

The outcome of acute leukemia patients with SET-NUP214 fusion after allogeneic stem cell transplantation.

SET-NUP214 fusion gene, also known as TAF-1-CAN and SET-CAN, is observed in acute myeloid leukemia (AML) and T-cell lymphoblastic leukemia (T-ALL). SET-NUP214 fusion in T-cell lymphoblastic leukemia is associated with chemotherapy resistance, but the prognosis of patients with AML with SET-NUP214 has rarely been reported. In the present study, we retrospectively analyzed all patients with acute leukemia including AML and T-ALL patients with SET-NUP214 fusion who underwent allogeneic stem cell transplantation (alloHSCT) in our center from July 2017 to November 2022. Of the total 11 patients, 5 patients were diagnosed with AML and 6 patients were diagnosed with T-ALL de novo. All patients received myeloablative regimens in CR1, and there were three (60%) AML patients who relapsed post-alloHSCT and three T-ALL (50%) patients who relapsed post-alloHSCT. Only one patient with AML who relapsed post-alloHSCT responded to subsequent chemotherapy plus donor lymphocyte infusion and survived the last follow-up. The estimated 1-year overall survival and 3-year overall survival for all these 11 patients were 69.3% and 38.5%, respectively. The estimated 1-year leukemia-free survival and 3-year leukemia-free survival for all patients were 69.3% and 38.5%, respectively. The research shows a high incidence of relapse for patients with acute leukemia with the SET-NUP214 fusion gene, even after alloHSCT. More clinical trials or research with larger samples are urgently needed for this group of patients.

Global, regional and national burden of endocrine, metabolic, blood and immune disorders 1990-2019: a systematic analysis of the Global Burden of Disease study 2019.

Background: Endocrine, metabolic, blood and immune disorders (EMBID) is a vital public health problem globally, but the study on its burden and global trends was scarce. We aimed to evaluate the global burden of disease and trends in EMBID from 1990 to 2019. Methods: We extracted the data of EMBID-related on death cases, Age-standardized death rates (ASDRs), disability-adjusted life-years (DALYs), Age-standardized DALY rates, years of life lost (YLLs), Age-standardized YLL rates, years lived with disability (YLDs) and Age-standardized YLD rates between 1990 and 2019 from the Global Burden of Disease 2019, by sex, age, and year at the global and geographical region levels. The Annual rate of change was directly extracted from Global Health Data Exchange (GHDx) and we also calculated the age-related age-standardized rate (ASR) to quantify trends in EMBID-related deaths, DALYs, YLLs and YLDs. Result: Globally, the EMBID-related ASDRs showed an increasing trend, whereas the DALYs ASR, YLLs ASR and YLDs ASR were decreased between 1990 to 2019. Furthermore, High-income North America and Southern Sub-Saharan Africa had the highest both ASDRs and DALYs ASR, and Southern Sub-Saharan Africa and Caribbean had the highest both YLDs ASR and YLLs ASR in 2019. Males had a higher EMBID-related ASDRs than females, but the DALYs ASR in females were higher than males. The burden of EMBID was higher in older-aged compared to other age groups, especially in developed regions. Conclusion: Although EMBID-related ASRs for DALYs-, YLLs- and YLDs declined at the global level from 1990 to 2019, but the ASDRs was increasing. This implied high healthcare costs and more burden of ASDRs due to EMBID in the future. Therefore, there was an urgent need to adopt geographic targets, age-specific targets, prevention strategies and treatments for EMBID to reduce negative health outcomes globally.

Bloodstream infections due to Carbapenem-Resistant Enterobacteriaceae in hematological patients: assessment of risk factors for mortality and treatment options.

PURPOSE: Bloodstream infection (BSI) caused by Carbapenem-Resistant Enterobacteriaceae (CRE) are associated with poor outcomes in hematological patients. The aim of this study was to identify risk factors for mortality and evaluate the value of epidemiological feature of carbapenemases in guiding antimicrobial treatment options. METHODS: Hematological patients with monomicrobial CRE BSI between January 2012 and April 2021 were included. The primary outcome was all-cause mortality 30 days after BSI onset. RESULTS: A total of 94 patients were documented in the study period. Escherichia coli was the most common Enterobacteriaceae, followed by Klebsiella pneumoniae. 66 CRE strains were tested for carbapenemase genes, and 81.8% (54/66) were positive, including NDM (36/54), KPC (16/54), IMP (1/54). Besides, one E. coli isolate was found to express both NDM and OXA-48-like genes. Overall, 28 patients received an antimicrobial treatment containing ceftazidime-avibactam (CAZ-AVI), of which 21 cases were combined with aztreonam. The remaining 66 patients were treated with other active antibiotics (OAAs). The 30-day mortality rate was 28.7% (27/94) for all patients, and was only 7.1% ((2/28) for patients treated with CAZ-AVI. In multivariate analysis, the presence of septic shock at BSI onset (OR 10.526, 95% CI 1.376-76.923) and pulmonary infection (OR 6.289, 95% CI 1.351-29.412) were independently risk factors for 30-day mortality. Comparing different antimicrobial regimens, CAZ-AVI showed a significant survive benefit than OAAs (OR 0.068, 95% CI 0.007-0.651). CONCLUSION: CAZ-AVI-containing regimen is superior to OAAs for CRE BSI. As the predominance of blaNDM in our center, we recommend the combination with aztreonam when choose CAZ-AVI.

Treatment With the Combination of Rituximab and Intravenous γ Globulin Is Effective in Promoting Engraftment in Donor-Specific Antibody Positive Patients Receiving Myeloablative Conditioning Haploidentical Stem Cell Transplantation.

Donor-specific anti-HLA antibodies (DSAs) are a major cause of engraftment failure in patients receiving haploidentical stem cell transplantation (HaploSCT). Effective procedures are needed for those who demand urgent transplantation and have no other donor options. We here retrospectively analyzed 13 patients with DSAs successfully treated with desensitization of rituximab and intravenous γ globulin (IVIg) before HaploSCT from March 2017 to July 2022. All 13 patients had DSA mean fluorescence intensity >4000 of at least 1 loci before desensitization. Of the 13 patients, 10 patients were with the initial diagnosis of malignant hematological diseases, and 3 were diagnosed with aplastic anemia. Patients were treated with 1 (n = 3) or 2 (n = 10) doses of rituximab (375 mg/m2 for 1 dose). All patients receive the same total dose of 0.4 g/kg of IVIg within 72 hours before haploidentical stem cell administration to neutralize the remaining DSA. All patients achieved neutrophil engraftment, and 12 patients achieved primary platelet engraftment. The patient with primary platelet engraftment failure received purified CD34-positive stem cell infusion nearly 1 year after transplantation and achieved platelet engraftment thereafter. The estimated 3-year overall survival is 73.4%. Although further studies on larger numbers of patients are needed, it is clear that the combination of IVIg and rituximab is an effective way to clear DSA and has a strong effect on promoting engraftment and survival for patients with DSA. It is a practical and adaptable combination of treatment options.

Venetoclax plus hypomethylating agents versus intensive chemotherapy for hematological relapse of myeloid malignancies after allo-HSCT.

Introduction: Since allogeneic stem cell transplantation (allo-HSCT) is considered one of the curative treatments for acute myeloid leukemia (AML) and myelodysplastic syndrome (MDS), hematological relapse following allo-HSCT remained a crucial concern for patients' survival. Methods: We retrospectively compared patients who received venetoclax plus hypomethylating agents (VEN+HMA, n=23) or intensive chemotherapy (IC, n=42) for hematological relapse of myeloid malignancies after allo-HSCT. HMA selection included decitabine (n=2) and azacitidine (n=21), and combined donor lymphocyte infusion was administered to 21 and 42 patients in VEN+HMA and IC groups, respectively. Results: Median age of all patients was 39 (16-64) years old. Overall response rates, including complete response (CR), CR with incomplete recovery of normal neutrophil or platelet counts (CRi) and partial response (PR), were not significantly different between VEN+HMA and IC groups (60.1% versus 64.3%, P=0.785). CR/CRi rate was 52.2% in VEN+HMA and 59.5% in IC group (P=0.567). The rate of relapse after response was 66.7% in VEN+HMA group and 40.7% in IC group (P=0.176). Median overall survival was 209.0 (95%CI 130.9-287.1) days for VEN+HMA group versus 211.0 (95%CI 28.7-393.3) days for IC group (P=0.491). The incidence of lung infection (17.4% versus 50.0%, P=0.010), thrombocytopenia (73.9% versus 95.2%, P=0.035) and acute graft-versus-host disease (aGvHD) (50.0% versus 13.0%, P=0.003) was significantly higher in IC group. Discussion: In conclusion, VEN+HMA is not inferior to IC regimen in terms of improving response and survival, and is associated with a lower incidence of adverse events and aGvHD. However, further research is required to enhance long-term survival.

Association between sociodemographic status and the T2DM-related risks in China: implication for reducing T2DM disease burden.

Objective: Analyzing the association between sociodemographic status and the type 2 diabetes mellitus (T2DM)-related risks in China to reduce the disease burden of T2DM. Methods: We downloaded data from the Global Burden of Disease Study 2019 to estimate the disease burden of T2DM in China. Secondary analyses were performed by year, age, gender, summary exposure value (SEV), and sociodemographic index (SDI). Results: In China, it is estimated that 3.74 (3.44-4.10) million incidence, 90.0 (82.3-98.5) million prevalence, 168.4 (143.2-194.0) thousand deaths, and 9.6 (7.6-11.9) million DALYs occurred in 2019, showing an increase of 96.8, 156.7, 162.8, and 145.4% compared to 1990. An inverse U-shaped curve was observed for the correlations between T2DM-related burden and SDI. A heavier burden was found in males. The top four risk factors were high body mass index (HBMI), dietary risks, air pollution and tobacco. HBMI, as the key risk, accounted for half of the disease burden of T2DM in China. Lower degree of SEV and higher level of attributable T2DM-related burden could be found in main risks, meaning their critical role of them in the development and progression of T2DM. An inverse U-shaped curve could be found in the association between age-standardized incidence, mortality, DALYs rate, and SDI. Conclusion: The disease burden of T2DM has rapidly increased in China. Gender disparities, different age distributions and inconsistent socioeconomic levels all played an important role in it. The key risk was HBMI. With the improvement of socioeconomic level, the main risk factors for T2DM have changed from environmental factors to lifestyle factors. Targeted control and preventative strategies to address adjustable risk factors could put an end to this soaring burden.

Clinical research advances of isavuconazole in the treatment of invasive fungal diseases.

Purpose: Invasive fungal diseases (IFD) are a major global public health concern. The incidence of IFD has increased the demand for antifungal agents. Isavuconazole (ISA) is a new triazole antifungal agent that has shown promising efficacy in the prophylaxis and treatment of invasive fungal diseases. The aim of this review is to summarize the recent real-world experiences of using ISA for the treatment and prevention of IFD. Methods: We performed a comprehensive literature search of the MEDLINE, PubMed, Embase, and Cochrane databases for clinical applications of ISA in the real world. Tables and reference lists are presented for this systematic review. Results: IFD poses a major threat to public health and causes high mortality rates. ISA may provide a good treatment. For example, the efficacy of ISA in the treatment of invasive aspergillosis (IA) is comparable to that of voriconazole, and its efficacy in the treatment of invasive mucormycosis (IM) is similar to that of liposomal amphotericin B (L-AmB); therefore, ISA is recommended as the first-line treatment for IA and IM. ISA can also achieve good efficacy in the treatment of invasive candidiasis (IC) and can be used as an alternative to de-escalation therapy after first-line drug therapy. In addition, most studies have shown the efficacy and safety of ISA for the prophylaxis of IFD. Conclusion: Taken together, ISA are expected to become a new choice for the treatment and prevention of IFD because of their good tolerability, high bioavailability, and few drug interactions.

Mucosal-associated invariant T cells predict increased acute graft-versus-host-disease incidence in patients receiving allogeneic hematopoietic stem cell transplantation.

BACKGROUND: Mucosal-associated invariant T (MAIT) cells are innate-like T cells, some studies have reported that the number of circulating MAIT cells reduced in patients with acute graft-versus-host-disease (aGVHD) development. However, the role of donor MAIT cells on aGVHD development and subsequent functional change still remain unclear. METHODS: The study recruited 86 patients with hematological malignancies who underwent allogeneic hematopoietic cell transplantation (HCT) from May 1, 2018 to June 30, 2019. MAIT cells, their subset, and cytokine levels were measured by flow cytometry. Gray's test was used to assess the impact of graft MAIT cell proportion and number on aGVHD incidence. The Cox proportional hazard model was used in the multivariate analysis. The comparison for continuous variables was assessed using Mann-Whitney analysis. RNA-sequencing was performed to investigate the possible molecular pathway changes. RESULTS: Our study showed that the proportion of MAIT cells in grafts was not different from normal controls, but the CD4/8 subsets were altered. Taking the median of the proportion and number of MAIT cells in the graft as the threshold, the results showed that the incidence of grade B-D aGVHD in patients with MAIT cell proportion ≥ 3.03% was significantly higher than that in patients with MAIT cell proportion < 3.03% (56.3%, 95% CI 37.1-71.2 versus 23.1%, 95% CI 13.8-46.2; P = 0.038).The number of MAIT cells in the graft was not associated with aGVHD development (P = 0.173), however, when the graft contained more CD4 positive, CD8 positive, and CD4/CD8 double-positive MAIT cells, the incidence of aGVHD was significantly increased (P = 0.019, P = 0.035 and P = 0.027, respectively). Besides, reduced frequencies and counts of circulating MAIT cells were observed in patients with aGVHD when compared to patients without aGVHD, accompanied by enhanced production of Tumor necrosis factor-α, Interferon-γ and upregulated programmed death-1, CXC Chemokine Receptor-6 (CXCR6) and CD38 expression. Gene set enrichment analysis of MAIT cell RNA-seq data showed interferon-α response pathway upregulated in aGVHD patients when compared with patients without aGVHD and healthy controls. CONCLUSIONS: Our study shows that MAIT cells in grafts and peripheral blood are both closely related to the aGVHD development post allogeneic HCT. Interferon-α response pathway perhaps is a critical regulation mechanism for the MAIT cell involvement in aGVHD development.

Ruxolitinib as an Effective and Steroid-Sparing First-Line Treatment in Newly Diagnosed BOS Patients After Hematopoietic Stem Cell Transplantation.

Bronchiolitis obliterans syndrome (BOS) is a life-threatening pulmonary complication of chronic graft-versus-host disease (cGVHD) after allogeneic hematopoietic stem cell transplantation (HSCT). In this study, we retrospectively identified seven patients newly diagnosed with BOS post HSCT and analyzed the outcomes in those patients treated with ruxolitinib as a first-line treatment. All seven patients achieved symptom responses within 2 weeks after ruxolitinib administration. Three months after treatment, five patients (71.43%) achieved a CR, and two (28.57%) achieved a PR. The overall response rate (ORR) was 100%. In addition, the steroid therapy was determined within 2 months after ruxolitinib treatment, indicating ruxolitinib as a steroid-sparing agent. We also found that ruxolitinib was well-tolerated and safe in treating newly diagnosed BOS. According to our results, ruxolitinib would be a promising and safe option in newly diagnosed BOS post HSCT.

The Global Burden of Osteoporosis, Low Bone Mass, and Its Related Fracture in 204 Countries and Territories, 1990-2019.

Background: Low bone mineral density (LBMD), including osteoporosis and low bone mass, has becoming a serious public health concern. We aimed to estimate the disease burden of LBMD and its related fractures in 204 countries and territories over the past 30 years. Methods: We collected detailed information and performed a secondary analysis for LBMD and its related fractures from the Global Burden of Disease Study 2019. Numbers and age-standardized rates related to LBMD of disability-adjusted life-years (DALYs) and deaths in 204 countries and territories were compared by age, gender, socio-demographic index (SDI), and location. Results: Global deaths and DALYs number attributable to LBMD increased from 207 367 and 8 588 936 in 1990 to 437 884 and 16 647 466 in 2019, with a raise of 111.16% and 93.82%, respectively. DALYs and deaths number of LBMD-related fractures increased 121.07% and 148.65% from 4 436 789 and 121248 in 1990 to 9 808 464 and 301 482 in 2019. In 2019, the five countries with the highest disease burden of DALYs number in LBMD-related fractures were India (2 510 288), China (1 839 375), United States of America (819 445), Japan (323 094), and Germany (297 944), accounting for 25.59%, 18.75%, 8.35%, 3.29%, and 3.04%. There was a quadratic correlation between socio-demographic index (SDI) and burden of LBMD-related fractures: DALYs rate was 179.985-420.435SDI+417.936SDI2(R2 = 0.188, p<0.001); Deaths rate was 7.879-13.416SDI+8.839 SDI2(R2 = 0.101, p<0.001). Conclusions: The global burden of DALYs and deaths associated with LBMD and its related fractures has increased significantly since 1990. There were differences in disease burden between regions and countries. These estimations could be useful in priority setting, policy-making, and resource allocation in osteoporosis prevention and treatment.

An Application of Analytic Hierarchy Process and Entropy Weight Method in Food Cold Chain Risk Evaluation Model.

The food cold chain is a special type of cold chain that refers to a system in which refrigerated and frozen food is always kept in the specified low-temperature environment in all links from production, storage, transportation, sales, distribution to consumption, so as to ensure food quality and to prevent food deterioration caused by temperature fluctuation. In recent years, the coronavirus disease 2019 (COVID-19) has brought a great impact on people's life and the social economy and also threatened the large-scale food cold chain. Through the effective identification and evaluation of high-risk factors in the food cold chain, this article has found the major risks that have a great impact on the entire food cold chain and proposes the specific measures of risk management and control to solve the problems of food cold chain and reduce risks quickly and efficiently to ensure the stability and safety of food cold chain and avoid the serious food safety accidents. The contribution of this article is reflected in three aspects, namely, (1) applies the expert system based on professional knowledge and rich experience and constructs a classification and identification system structure of food cold chain risk indexes, which lay a foundation for further identifying and evaluating the major risks of the food cold chain; (2) designs a comprehensive index weighting method combining the AHP method and entropy weight method to quantitatively evaluate the major risks. This comprehensive method combines a hierarchical structure system, evaluation algorithm, subjective factor correction algorithm, and so on. The evaluation results are more accurate, have a high matching degree with reality, and have good theoretical and practical significance; (3) analyzes and explains the major risks of the food cold chain in the non-epidemic situations and COVID-19 situations. Proposals and measures for risk management and control are put forward, which have wide practical significance.

Development of Mini-Barcode Based on Chloroplast Genome and Its Application in Metabarcoding Molecular Identification of Chinese Medicinal Material Radix Paeoniae Rubra (Chishao).

Radix Paeoniae Rubra (Chishao), a typical multi-origin Chinese medicinal material, originates from the dried roots of Paeonia lactiflora or P. veitchii. The previous study suggested that these two commonly used Chishao showed variation in their chemical compositions and clinical efficacies. Therefore, accurate identification of different Chishao species was of great significance for the guide of clinical medication, and timely treatment of patients. In this study, the chloroplast genome sequences of P. lactiflora and P. veitchii were obtained by next-generation sequencing (NGS) technology, and then the hypervariable regions were selected to design two mini-barcode candidates for species identification. Combined with DNA metabarcoding technology, we performed qualitative and quantitative analysis on the artificially mixed samples of P. lactiflora and P. veitchii and evaluated the identification ability of these mini-barcode candidates. Furtherly, the mini-barcode with good performance was applied to distinguish the Chinese patent medicine "cerebral thrombosis tablets" containing Chishao. The results indicated that the chloroplast genomes of P. lactiflora and P. veitchii were 152,750 and 152,527 bp, respectively. As published previously, they exhibited a typical quadripartite structure including a large single-copy region (LSC), a small single-copy region (SSC) and a pair of inverted repeat regions (IRs). The nucleotide polymorphism analysis revealed seven variable protein-coding regions as petL, psaI, psbJ, rpl16, ycf1b, psaC, and ndhF, and two mini-barcodes were developed from ycf1b and ndhF respectively. The result suggested that both two mini-barcodes performed well distinguishing P. lactiflora from P. veitchii. Besides, P. lactiflora was the only raw material of Chishao in all collected "cerebral thrombosis tablets" samples. In general, this study has established a method to realize the qualitative and quantitative identification of Chishao as multi-origin Chinese medicinal materials, which can be applied to Chinese patent medicines containing Chishao.