Current Biotechnology

Progress of CRISPR/Cas9 Application in Wheat Breeding
Qiao CAO, Zhanliang SHI, Guocong ZHANG, Jinfu BAN, Shusong ZHENG, Xiaoyi FU, Shichang ZHANG, Mingqi HE, Ran HAN, Zhenxian GAO
2021, 11(6): 661-667. DOI: 10.19586/j.2095-2341.2020.0104
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Triticum aestivum L. is one of the main crops in the world and plays an important role in food security supply. In the past few decades， due to the complexity of wheat genome and the difficulty of genetic transformation， the basic and applied research of wheat has lagged behind other cereal crops. In 2014， remarkable progress was made in wheat genome editing， which promoted the development of wheat biotechnology. This paper summarized the research progress of CRISPR/Cas9 technology in wheat breeding， briefly introduced the discovery， principle， advantages and disadvantages of CRISPR/Cas9 gene editing technology， and pointed out that the Agrobacterium mediated genetic transformation in the process of wheat gene editing would reduce the gene silencing frequency compared with particle bombardment， and would become the mainstream genetic transformation in the process of gene editing in the future； optimizing the promoter of sgRNA and selecting the conserved sequence of homologous gene as the target can improve the efficiency of gene editing； the newly developed base editor and prime editor need to introduce more mutation types， and the feasibility of further improving the efficiency and safety of wheat gene editing was prospected to provide reference for future wheat breeding work.

Research Progress of Tocotrienol
Yanjiao LI, Yuan GAO, Lei WANG, Lan ZHANG
2021, 11(6): 668-675. DOI: 10.19586/j.2095-2341.2021.0167
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Tocotrienol and tocopherol collectively referred to as vitamin E， are important fat?soluble vitamins. Vitamin E can only be synthesized by plants or photosynthetic bacteria. It is essential for human and animal nutrition and can only be ingested from outside. For a long time， due to its lower bioactivity and limited distribution than tocopherol， few research has been done on tocotrienol. Recent studies have found that due to the similar structure of tocotrienol and tocopherol， tocotrienol has the same antioxidant as tocopherol. While due to the unsaturated side chain， tocotrienol also has some functions different from tocopherol， such as protecting nerves from damage， lowering cholesterol and protecting brain cells from damage. Therefore， the research of tocotrienol has gradually become a hot spot. According to the biosynthetic pathway of vitamin E， people have also begun the study of tocotrienol biofortification， overexpression of the first key enzyme gene HGGT is the most effective way to improve the content of tocotrienol. In the future， it is necessary to combine the molecular mechanism of its synthesis and regulation and its absorption and utilization to develop functional products targeting at tocotrienol. In this paper， the synthetic pathway， biological function and biofortification of tocotrienol were reviewed， and the research focus in the future was prospected.

Overview and Progress of Safety Regulatory System of Genetically Modified Crops in Argentina
Mengjie LI, Xiaoyun HE, Tao TONG, Jingang LIANG, Kunlun HUANG
2021, 11(6): 676-687. DOI: 10.19586/j.2095-2341.2021.0122
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Argentina is one of the lead countries to adopt genetically modified crops and is currently the third largest grower of genetically modified crops in the world. Argentina has been one of the pioneers in the world， especially Latin American countries in regulation and approval of biotechnology products. Its rich experience in the regulation of genetically modified crops and the recognition of the Food and Agriculture Organization of the United Nations have made Argentina as one of the global leaders in the regulation of genetically modified crops. This article introduced the research and application of genetically modified crops， the regulatory system of genetically modified crops and new breeding technologies， the import and export situation as well as the traceability system of genetically modified crops in Argentina， and discussed the impact of the introduction of genetically modified technologies on this country， aiming to comprehensively understand the regulatory system of genetically modified crops and new breeding technologies in Argentina， and provide a reference for the safety management of genetically modified crops for China.

The Application Potential of Mesenchymal Stem Cells in the Treatment of Diseases
Hui WANG, Pengxiang ZHAO, Xujuan ZHANG, Tingting YAO, Mengyu LIU, Mawulikplimi Adzavon YAO, Fei XIE
2021, 11(6): 688-693. DOI: 10.19586/j.2095-2341.2021.0055
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Mesenchymal stem cells （MSCs） are pluripotent cells derived from the mesoderm and can be differentiated into multiple mesenchymal cell lineages， and can produce a variety of interfilled mass spectrometry systems， including osteoblasts， adipocytes， chondrocytes and muscle cells. MSCs also have the ability to secrete a variety of cytokines， which can promote angiogenesis， epithelial regeneration and so on， which have great potential for regenerative medicine. Studies have confirmed that MSCs can promote tissue regeneration and accelerate wound healing by differentiating into multiple cell types； improve tissue fibrosis by secreting cytokines； and can induce tumor cell apoptosis by carrying drugs， thereby inhibit tumor development. However， the potential of MSCs to differentiate into fibroblasts and the ability to promote tumor growth decrease the safety of MSCs in clinical treatment. We summarized the role of MSCs in the occurrence and development of tumors， chronic wounds， fibrosis?related diseases， cardiovascular diseases and other diseases， and the clinical application of MSCs. The potential application value of MSCs in the treatment of related diseases and the challenges were further discussed， in order to provide reference for the clinical application of mesenchymal stem cells.

Application Progress of Metagenomic Technology in Acne Research
Wei LI, Chong WANG, Sijia LIU, Minyi YANG, Yunping ZHANG
2021, 11(6): 694-699. DOI: 10.19586/j.2095-2341.2021.0073
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The development of high?throughput sequencing technology has improved the people's understanding of the microbiome. Metagenomics technology is widely used in the research of various environmental microbiomes due to its comprehensive and in?depth analysis function， especially in clarifying the relationship between various diseases and the human microbiome， metagenomics technology plays an important role. As a common skin disease， acne seriously affects people's skin beauty and mental health. The use of metagenomics technology to explore the relationship between skin microbiome and acne will help the research on the pathogenesis of acne and the improvement of clinical treatment methods. By introducing the development background， overview and application research progress of metagenomics technology， the paper explored the relationship between skin microbes and acne， reviewed the application status of metagenomics technology in acne research， and summarized the current metagenomics technology in skin disease research， and the existing problems of metagenomics technology in the research of skin diseases， aiming to provide reference for the research of metagenomics of acne.

Research Progress on the Pathogenesis of Diabetic Cardiomyopathy
Min LIAO, Luo YANG, Zhen WANG, Yarong HAO
2021, 11(6): 700-704. DOI: 10.19586/j.2095-2341.2021.0023
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Diabetic cardiomyopathy （DCM） is a special type of heart disease， which increases the risk of heart failure in diabetic patients to some extent and is also one of the main causes of death in diabetic patients. The pathogenesis of DCM involves many aspects. Myocardial cell metabolic disorders （such as hyperglycemia and insulin resistance）， myocardial inflammation and fibrosis are the basis of the pathogenesis of DCM， and these factors act on the occurrence and development of DCM individually or in combination. At present， there is no effective drug for the radical cure of DCM in clinical practice， and the study of the pathogenesis of DCM is of great significance in the development of targeted therapy drugs. The pathogenesis of DCM was mainly reviewed in order to provide theoretical basis for the early prevention and treatment of DCM.

Research Progress of Temozolomide in the Mechanism of Drug Resistance in Glioblastoma
Tingting YAO, Mengyu LIU, Pengxiang ZHAO, Hui WANG, Yang YI, Fei XIE, Mawulikplimi Adzavon YAO
2021, 11(6): 705-710. DOI: 10.19586/j.2095-2341.2021.0022
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Glioblastoma is the most malignant primary brain tumor in glioma， which has the characteristics of low cure rate， high recurrence rate and invasive growth. Without the use of chemotherapy drugs， the median survival time of patients is only 12.1 months. The standard treatment for patients with glioblastoma is surgical resection supplemented by chemoradiotherapy. Among them， temozolomide （TMZ） as a new oral alkylating agent， is the first?line drug in the current chemotherapy of glioma. However， median survival improved by only 2 months after temozolomide treatment， mainly because glioblastomas can develop resistance to TMZ. Glioblastoma on TMZ resistance mechanism is mainly DNA repair mechanisms， it includes O⁶?methyl guanine DNA methyltransferase （MGMT） of sites for the action of the drug directly repair， mismatch repair （MMR） and base excision repair （BER）， these repair mechanisms can repair DNA damage cause by TMZ， thereby reducing tumor cell sensitivity to TMZ. The research progress on the mechanism of TMZ drug resistance of glioblastoma in recent years was reviewed， in order to provide a theoretical basis for the development of new therapeutic methods.

Progress on Correlation Between Insulin Growth Factor 2 mRNA Binding Protein 3 and Tumor
Zihao GUO, Tiemin PEI, Desen LIANG, Fan YUAN
2021, 11(6): 711-717. DOI: 10.19586/j.2095-2341.2020.0162
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RNA binding protein is an important regulator in the life cycle of organisms. As one of the RNA binding proteins， insulin growth factor 2 mRNA binding protein 3 （IGF2BP3） mediates the occurrence and development of cancer cells as well as the prognosis and outcome of diseases. Therefore， it is great significance to explore the mechanism of IGF2BP3 carcinogenesis， the mechanism of pathway conduction， and the therapeutic target based on IGF2BP3 molecule. To elaborate the biological functions of the IGF2BPs family， the relevance of IGF2BP3 in some tumors and diseases， the application prospects of IGF2BP3 in modern clinical treatment， we aimed to provide a theoretical basis for further research on the IGF2BP3 gene.

Effects of AMPK/SIRT1/PPARγ/PGC1α Axis and Related Factors on Lipid Metabolism in Osteoarthritis
Chuancai LIANG, Peng YI, Bo QIU
2021, 11(6): 718-723. DOI: 10.19586/j.2095-2341.2021.0019
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Osteoarthritis （OA） is a degenerative joint disease with cartilage degeneration， osteosclerosis， and chronic synovial inflammation as the main clinical features. In the pathological changes of osteoarthritis， abnormal lipid metabolism is closely related to the degenerative changes of cartilage and bone. After the activation of adenosine monophosphate?activated protein kinase （AMPK）， which can regulate the lipid metabolism of chondrocytes by regulating the key enzymes of fatty acid synthesis carnitine palmitoyltransferase 1（CPT?1）， medium chain acyl?CoA dehydrogenase （MCAD） and chondrocyte autophagy to delay the development of OA. Peroxisomal proliferator?activated receptor γ （PPARγ） and peroxisomal proliferator?activated receptor γ coactivator 1α （PGC?1α） also have similar physiological functions.The activation and interaction of AMPK and silencing information regulator 1 （SIRT1） can mediate the signal transduction and physiological functions of PPARγ and PGC?1α. This paper summarized the role of AMPK/SIRT1/PPARγ/PGC1α axis in the pathogenesis of OA， which was expected to provide reference for therapy and prevention research of OA.

Progress on the Production Technology of Recombinant Therapeutic Proteins
Nana ZHOU, Xiaoyan WANG, Yuan ZHANG, Jing WANG, Guomiao ZHAO, Chao WEI, Kai YANG, Tai AN
2021, 11(6): 724-731. DOI: 10.19586/j.2095-2341.2021.0130
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Recombinant therapeutic proteins are the core products in biopharmaceuthc. They are mainly used to product functional proteins or their mutants by genethcally engineered bacteria to compensate for the loss of protenis in the body and play a key role in the treatment of diseases. In recent years， recombinant therapeutic proteins are playing an increasingly important role in the treatment of diseases， and related technologies are developing rapidly. The middle and upstream production process of recombinant therapeutic proteins was reviewed. The latest technological progress of recombinant therapeutic proteins in expression system， cell culture， purification and quality control was analyzed. The technological advances in the preparation of recombinant therapeutic proteins were demonstrated. It was expected to provide certain reference for domestic production of recombinant therapeutic proteins.

Establishment and Application of CRISPR/Cpf1‑mediated Base Editing System
Shulei LI, Miaoyun XU, Hongyan ZHENG, Lei WANG
2021, 11(6): 732-740. DOI: 10.19586/j.2095-2341.2021.0088
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The excellent traits of crops often come from single base mutations in their corresponding genes， and traditional breeding cannot easily obtain such targeted base mutations. The base editing technology is a gene editing technology based on the clustered regularly interspaced short palindromic repeats/CRISPR?associated proteins （CRISPR/Cas） system， which can perform targeted replacement of specific bases on the target sequence without causing DNA double?strand break. In order to expand the recognition range of base editing technology in crops， the FnCpf1 from Francisella novicida and the cytosine deaminase APOBEC1 were used to improve the base editing system. Then， constructing the corresponding vector targeting to the maize BT2 gene. Detected its editing ability by means of instantaneous conversion. The test results found 9 types of base changes. Among them， the cytosine of the 11th base at the 5' end of the target site was converted to adenine， and the base editing efficiency reached 2.5%. It showed that the system can recognize "TTN" as a protospacer?adjacent motif （PAM） site and single?base editing of the target site， which can provide research ideas for the expansion of the recognition range of base editing.

Construction and Application of Plasmid‑based and Xylose‑induced Transformation System of Bacillus subtilis
Hui TIAN, Shuai WANG, Bo LIU
2021, 11(6): 741-748. DOI: 10.19586/j.2095-2341.2021.0044
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Bacillus subtilis is a kind of food?grade safe strain. Due to clear physicochemical characteristics and simple fermentation， it is widely used in the high?efficiency expression of heterologous proteins and the synthesis of high value?added compounds. The traditional genetic transformation methods for B. subtilis have some demerits， such as complicated operation procedures and low efficiency. It is significant important to develop a convenient and highly efficient genetic transformation system. The transcription factor ComK was proved to regulate the formation of competent cells and play an important role in highly efficient transformation of B. subtilis. A shuttle plasmid pUBC01?P_xyl?comK containing a xylose?induced P_xyl?comK expression cassette was constructed and a recombinant strain B. subtilis K1 containing this plasmid was developed. The transformation efficiency of plasmid pHY300?p43?egfp into B. subtilis K1 could reach 4.8×10³ CFU·μg^-1 after optimized xylose induction condition. In addition， pUBC01?P_xyl?comK could be cured via serial subculture under nonselective condition. The system of xylose?induced competent cells and plasmid curing greatly increased the convenience of gene editing and strain improvement of B. subtilis and enhanced character stability of the transformed strains， especially production strains.

Global Patent Analysis of Lipase Development Trends
Yage ZHANG, Yu PANG, Wei ZHANG, Zhengfu ZHOU
2021, 11(6): 749-757. DOI: 10.19586/j.2095-2341.2020.0161
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Lipase is a kind of green and environment?friendly biocatalyst which can catalyze the break and formation of various ester bonds. Lipases have been widely used in many fields such as food， feed， detergent， pharmaceutical， fine chemistry and bioremediation. Based on Pat Snap， statistic on lipase?related patents in various countries and regions around the world before October 2020， we analyzed the lipase technology distribution， development status and future trends， and comparative analyzed the lipase research and development in China to provide a reference for the development of lipase technology and industry.

Analysis on Changes of Volatile Flavor Compounds in Ruan′er Pear Wine During Fermentation
Pengzheng PEI, Jianmin YUN, Qi JIA, Yuxuan ZHAO, Lin HAO
2021, 11(6): 758-769. DOI: 10.19586/j.2095-2341.2021.0086
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Ruan'er pear is a unique "frozen fruit" product in Northwest China. It is sour， sweet and delicious with rich fruit aroma. It is the best raw material for brewing fruit wine. In order to clarify the retention of fruit aroma in the fermentation process of fruit wine， and to explore the effect of fermentation time on the quality of Ruan'er pear wine， the frozen fruit of Ruan'er pear in Minhe County of Qinghai Province was used as raw material. Based on the fermentation process with skin residue， the volatile flavor compounds in raw juice （0 d）， pre fermentation （7， 14， 28 d） and post fermentation （100 d） samples were analyzed by headspace solid?phase microextraction?gas chromatography?mass spectrometry （HS?SPME?GC?MS）. Effects of original fruit flavor retention and post fermentation time on flavor of Ruan'er pear wine were discussed by relative odor activity value （ROAV） and principal component analysis （PCA）. The results showed that： a total of 88 volatile compounds were detected in the whole fermentation process， including 33 esters， 29 alcohols， 6 acids， 7 terpenes and 13 other compounds， and the total content of flavor compounds increased first and then decreased slightly with the fermentation process. The results of ROAV showed that there were 8 kinds of key flavor compounds in original fruit of Ruan'er pear， which were ethyl butyrate， ethyl hexanoate， ethyl 2?methylbutyrate， n?octanol， damascone， linalool， eugenol and decanal； among them， ethyl caproate， n?octanol， linalool， eugenol and damascone were well preserved in fruit wine， which were the key substances to determine the flavor of Ruan'er pear wine. Principal component analysis showed that the main aroma contributing substances in the early stage of fermentation were ethyl butyrate， 3?hydroxyethyl butyrate and 2?methylethyl butyrate. When fermented to 100 d， ethyl acetate， ɑ?terpineol， limonene and linalool contributed a lot to the aroma. These aroma compounds together gave Ruan'er pear wine unique flavor quality of delicate fragrance and rich fragrance. In this study， the key flavor compounds of Ruan'er pear wine and the change rules of characteristic flavor compounds in different fermentation stages were obtained， which can provide theoretical support and reference for the development of high?quality Ruan'er pear wine products and the improvement of Ruan'er pear wine fermentation process.

Prokaryotic Expression and Immunogenicity Detection of the H Protein of Peste des Petits Ruminants Virus
Dan LI, Haozhi SONG, Weisong GAO, Xingjian LIU, Zhifang ZHANG, Yinyu LI
2021, 11(6): 770-776. DOI: 10.19586/j.2095-2341.2021.0039
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Peste des petits ruminants is an acute and contagious infectious disease caused by the peste des petitis ruminant virus （PPRV）， which has a serious impact on the development of China's animal husbandry. At present， vaccination is the main method of prevention and control. In order to detect the main antigen protein hemagglutinin （H） protein of PPRV， searched its amino acid sequence published in recent years from the GenBank databases， and selected a sequence that conformed to the popular trend in China. The gene sequence was optimized and synthesized， and then cloned into the pET28a vector. The high?expressing Rosetta （DE3） strain was selected for expression and the product was identified by SDS?PAGE， Western Blot and mass spectrometry analysis. A single H protein was purified by nickel column affinity chromatography， and 20 μg H protein was mixed with adjuvant to immunize mice， and serum was collected for antibody titer detection. The results showed that the serum antibody titer reached 1：6 400 after two weeks of the second immunization， indicating that H protein has good immunogenicity. Further testing of neutralizing antibodies on this serum showed that it had a neutralizing effect on the PPRV vaccine strain， and the neutralizing titer did not exceed 1：40. The results provide a theoretical basis for the use of H protein in the development of PPRV vaccines.

Research on Functional Evaluation Method of Enzymes in Nucleic Acid Detection Kits
Zhenzhou YANG, Yan LI, Xue YANG, Gang LIU, Wen LIANG
2021, 11(6): 777-782. DOI: 10.19586/j.2095-2341.2021.0105
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The performance of enzyme in severe acute respiratory syndrome coronavirus 2 （SARS?CoV?2） nucleic acid detection kit plays a decisive role in the detection effect. In order to explore the rapid evaluation method for the performance of reverse transcriptase and DNA polymerase， the technical properties of reverse transcriptase and DNA polymerase in the common SARS?CoV?2 nucleic acid detection kit were comprehensively evaluated based on the reference material of SARS?CoV?2 nucleic acid. In the first step， the reverse transcription efficiency of reverse transcriptase was investigated by analyzing the concentration ratio of cDNA product to target RNA. In the second step， the same cDNA was used as the target sample for real?time fluorescence quantitative PCR analysis to compare the amplification efficiency of DNA polymerase. In the third step， using COVID?19 RNA standard substance as target sample， reverse transcription PCR （RT?PCR） was used to compare the amplification efficiency of the kit. In the fourth step， to compared the amplification efficiency of DNA polymerase. The results showed that the reverse transcription efficiency of the tested kits ranged from 6.5% to 80.7%， the amplification efficiency of DNA polymerase ranged from 90.4% to 119.7%， and the amplification efficiency of the total reverse transcription PCR ranged from 90.7% to 102.4%. The thermal stability evaluation results showed that the enzyme activity， which was initially very good， may change in quality during storage and transportation. By reflecting the quality level of enzyme in SARS?CoV?2 nucleic acid detection kit， it has certain reference value for relevant quality control work.

Analysis on the Research Status of Aptamer Technology Based on CiteSpace
Yongyi LIANG, Yue LONG, Jiahua XIAN, Jun LIN, Jin PENG
2021, 11(6): 783-794. DOI: 10.19586/j.2095-2341.2021.0127
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To objectively and comprehensively display and analyze the research status， research hotspots and development trend of aptamer by using visual analysis software， so as to provide guidance and reference for the research of aptamer. Using the Web of Science platform database as the data source， CiteSpace 5.5.R2 software was used to analyze the literature related to aptamer from January 1st， 1900 to December 31th， 2020. A total of 14 939 articles were included. WEIHONG TAN， KHALIL ABNOUS and other authors published the most papers in this field. In recent years， the emerging terms were “wide linear range”“pg ml”“clinical diagnosis”“early diagnosis”， etc， these terms are the research hotspots in this field. The author co?citation analysis and literature co?citation analysis showed that the research in this field mainly focused on the in vitro screening of aptamers， the change of aptamer conformation， the application of aptamers in drug targeted therapy and aptamer biosensors. CiteSpace software was used to analyze the published aptamer literatures in Web of Science database， revealing the research hotspots in this field， from in vitro screening and aptamer conformation change （1992—2002） to the application of aptamers in drug targeted therapy and aptamer biosensor （2002—2020）. Combined with the emerging terms in recent years， it is predicted that the application of aptamer in drug targeted therapy and aptamer biosensor will continue to be the future research direction.

Detection the Affinity Between PDL1 Antibodies and PDL1 Antigen Based on Biolayer Interferometry
Yan HU, Ling CHEN
2021, 11(6): 795-801. DOI: 10.19586/j.2095-2341.2021.0003
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Biolayer interferometry （BLI） technology can conduct a comprehensive analysis of affinity and kinetics of the interaction between antibody and antigen. There is a large demand for streptavidin （SA） biosensors in antibody clone screening and determination of kinetic constants， but there are few reports on the reuse of SA sensors. Based on BLI technology and regenerative SA biosensor， a method for detecting the affinity of PDL1 antibody and PDL1 antigen using the regenerated sensor was established. By coupling the biotinylated PDL1 antigen to the SA biosensor， and then combining with the four types of PDL1 antibodies， single?chain antibody， bivalent single?chain antibody， intact antibody， and bispecific antibody， to calculate the affinity constant of the antigen?antibody， using glycine （10 mmol·L^-1， pH 1.7） regenerated the SA sensor， and analyzed the intermolecular interaction force again. The result showed that the mean repeatability relative standard deviation （RSD） was 6.87%， inter?batch repeatability RSD was 0.82%， and the mean stability RSD was 6.13%， the data showed that the SA biosensor regenerated by glycine had reliable data， good reproducibility and high stability， and reused SA sensor could still continue to be used for real?time， lable?free antigen?antibody interaction analysis of previous sample. BLI technology can greatly save the detection cost and provide the experimental basis for the repeated use of SA sensor.

The Effect of Galectin‑3 on the Abnormal Mineralization of Osteoarthritic Osteoblasts
Yong HU, Jian YANG, Bo QIU
2021, 11(6): 802-806. DOI: 10.19586/j.2095-2341.2021.0031
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Inflammation is represented by inflammatory factors， which is an important mechanism for the pathogenesis of subchondral bone in osteoarthritis （OA）. The inflammatory factor galectin?3 （Gal?3） can induce OA lesions of subchondral bone， but the mechanism is still unclear. We used tibial plateau specimens from patients who received total knee arthroplasty due to knee joint OA to culture osteoblasts. After treating osteoblasts with Gal?3 under normoxia and hypoxia， respectively， the expression of osteocalcin， ERRα and Sirtuin1 were detected. Meanwhile， the osteoblasts cultured in the mineralization solution were treated with Gal?3， and after 28 days， the degree of mineralization of osteoblasts was detected by alizarin red staining and quantitative analysis was performed. The results showed that Gal?3 inhibited the expression of osteocalcin in osteoblasts， induced the expression of ERRα and Sirtuin1 under hypoxic conditions， and Gal?3 promoted the mineralization of OA osteoblasts under normal oxygen conditions. Under normal oxygen conditions， Gal?3 can induce abnormal mineralization of OA osteoblasts， and under hypoxic conditions Gal?3 promotes osteogenesis， indicating that Gal?3 plays an important role in the mineralization of OA osteoblasts.

Analysis of Improvement Effect and Mechanism of Tenuigenin and Liensinine on Sevoflurane‑induced Acute and Longterm Cognitive Impairments of Neonatal Rats
Lili HAN, Huixia WANG, Xinang LAN, Huimei HAN, Hongchao SUN
2021, 11(6): 807-812. DOI: 10.19586/j.2095-2341.2021.0013
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The aim of this study was to analyze the improvement effect and mechanism of tenuigenin and liensinine on sevofluraneinduced acute and longterm cognitive impairments of neonatal rats. 36 neonatal SD rats were randomly divided into TCM （traditional Chinese medicine） group （n=12）， model group （n=12） and blank group （n=12）. TCM group and model group were exposed to 2.5% sevoflurane environment for 2 h·d^-1 of anesthesia induction and lasted for 3 days， and blank group was exposed to the same environment without sevoflurane. TCM group was intraperitoneally injected with a 1：1 compatible solution of tenuigenin and liensinine （30 mg·kg^-1） at 30 min before anesthesia， and model group and blank group were injected intraperitoneally with the same amount of 0.9% NaCl solution. The blood gas indicators， cortical electroencephalogram results， Morris water maze behavior and serum corticosterone content were compared among groups. Western blot was used to detect the expression levels of Bcl₂， Bax and Caspase?3 in hippocampus. The total duration， average duration and quantity of epileptic waves of cortical electroencephalogram in TCM group were lower than those in model group （P<0.05）. Results of positioning navigation experiment showed that the escape latency in TCM group was shorter than that in model group （P<0.05）. Results of spatial probe test showed that the exploration time in TCM group was longer than that in model group （P<0.05）. Serum corticosterone content of rats in model group was higher than that in blank group （P<0.05）， and the content in TCM group was lower than that in model group （P<0.05）. The expression level of Bcl₂/Bax in hippocampus of TCM group was higher than that of model group （P<0.05）， while the expression level of cleaved Caspase?3 was lower than that of model group （P<0.05）. Results showed that tenuigenin and liensinine can improve the sevofluraneinduced acute and longterm cognitive impairments in neonatal rats. The mechanism may be related to regulating the expressions of apoptosisrelated genes Bcl₂， Bax and Caspase?3.

Singapore Agricultural Biotechnology Annual (2020)
Di WANG, Chaohua MIAO, Wujun Compiler JIN
2021, 11(6): 813-817. DOI: 10.19586/j.2095-2341.2021.0077
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Singapore does not have any domestic commercial production of plant biotechnology. As of October 2020， a total of 43 genetically engineered （GE） plant products have been approved for use as food or food ingredients in the country. Singapore’s Genetic Modification Advisory Committee （GMAC） recently revised its regulations on stacked events to adopt the "high covers low" approach which exempts higher order combinations of stacked events from regulatory assessment if they are derived from prior GMAC?endorsed lower order combinations.

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