High-throughput screening of mouse gene knockouts identifies established and novel skeletal phenotypes

Screening gene function in vivo is a powerful approach to discover novel drug targets. We present high-throughput screening （HTS） data for 3 762 distinct global gene knockout （KO） mouse lines with viable adult homozygous mice generated using either gene-trap or homologous recombination technologies. Bone mass was determined from DEXA scans of male and female mice at 14 weeks of age and by microCT analyses of bones from male mice at 16 weeks of age. Wild-type （WT） cagemates/littermates were examined for each gene KO. Lethality was observed in an additional 850 KO lines. Since primary HTS are susceptible to false positive findings, additional cohorts of mice from KO lines with intriguing HTS bone data were examined. Aging, ovariectomy, histomorphometry and bone strength studies were performed and possible non-skeletal phenotypes were explored. Together, these screens identified multiple genes affecting bone mass： 23 previously reported genes （Calcr, Cebpb, Crtap, Dcstamp, Dkkl, Duoxa2, Enppl, Fgf23, Kissl/Kisslr, Kl （Klotho）, Lrp5, Mstn, Neol, Npr2, Ostml, Postn, Sfrp4, S1c30a5, Sic39a13, Sost, Sumf1, Src, Wnt10b）, five novel genes extensively characterized （Cldn18, Fam20c, Lrrkl, Sgpll, Wnt16）, five novel genes with preliminary characterization （Agpat2, RassfS, Slc10a7, Stc26a7, Slc30a10） and three novel undisclosed genes coding for potential osteoporosis drug targets.

Bioinformatics Analysis Raises Candidate Genes in Blood for Early Screening of Parkinson's Disease

Parkinson＇s disease （PD） is a typical degenerative disease, which is characterized by the most obvious symptoms of movement dysfunction, including shaking, rigidity, slowness of movement and difficulty in walking and gait. This disease can not be clearly identified through laboratory tests at present, thus application of high-throughput technique in studying the expression profiles of PD helps to find the genetic markers for its early diagnosis. Studies on expression profiles of neurodegenerative diseases have revealed the novel genes and pathways involved in the progress of illness. In this study, the expression profiles of PD in blood were compared, showing that 181 differentially expressed genes （DEG） exhibit a similar expression trend both in patients and in normal controls.

Wolman Disease in Bulgarian Patients: Selective Genetic Screening in Two Presumable Endemic Regions

Wolman disease is a rare autosomal recessive disorder caused by mutations in the LIPA gene (10q23.31). The LIPA gene encodes lysosomal acid lipase (LAL), which plays a key role in hydrolysis of the cholesteryl esters and triglycerides. Two unrelated families from Bulgaria were referred for genetic testing with clinical diagnosis Wolman disease. Sanger sequencing of all coding exons and exon-intron boundaries of the LIPA gene was performed. The index patients were found to be homozygous for two different mutations in the LIPA gene: a missense mutation, c.260G > T, p.Gly87Val, which affects the enzyme active site and a splice-site change, c.822+1G > A, which most probably destroys the enzyme polypeptide chain. These two completely different types of mutations along the LIPA gene resulted in a very similar phenotype involving liver, kidney, gastrointestinal, muscle and blood disturbances. As consanguinity is not typical for the Bulgarian population, a possible explanation of the homozygosity could be presence of endemic regions for given mutations. To check this hypothesis, selective screening for these mutations was performed in two presumable endemic regions in Bulgaria. Altogether, 100 newborns were screened for p.Gly87Val mutation and the detected carrier frequency was about 1% (1/100), while in the group of 100 newborns screened for the c.822 + 1G > A mutation the detected carrier frequency was 2% (2/100). The results indicate a high recurrence risk of Wolman disease in these particular Bulgarian regions of about 1:10000. These findings are from crucial importance for the inhabitants of the corresponding parts of Bulgaria. They may benefit from early genetic testing and adequate genetic counselling during family planning.

Tay-Sachs carrier screening in the genomics age: Gene sequencing versus enzyme analysis in non-Jewish individuals

Purpose: To compare the sensitivity of Hexosaminidase A (HexA) enzyme-based testing to gene sequencing for carrier detection in non-Jewish individuals. Methods: Blood samples were obtained from parents and relatives of affected patients at an annual Tay-Sachs and Allied Diseases Foundation meeting. A family history was taken for each individual. Samples were analyzed for leukocyte HexA activity, serum HexA activity and subjected to extensive gene sequencing. The results from these analyses were combined with our previously published data describing 34 obligate Tay-Sachs disease (TSD) carriers. Results: Twelve additional TSD carriers were detected in this study. Gene sequencing successfully identified all 12 carriers whereas enzyme analysis identified 11 of 12 carriers. This individual is a carrier of the B1 variant that is known to cause false negative results with enzyme testing. Combined data from 46 non-Jewish TSD carriers revealed that gene sequencing had a higher sensitivity rate than HexA enzyme-based testing (94% versus 87%) in non-Jewish TSD carriers. In our series, approximately 4% of non-Jewish TSD carriers have this mutation. Conclusions: HexA gene sequencing provides a higher sensitivity for TSD carrier detection than HexA based enzyme analysis in non-Jewish patients primarily due to the presence of individuals with the B1 variant.

Cervical cancer screening: hTERC gene amplification detection by FISH in comparison with conventional methods

Aim: To assess the clinical significance of hTERC amplification for cervical cancer screening detected by fluorescence in situ hybridization (FISH) and compare it with that of current screening methods within the same group. Methods: A total of one hundred and nine women were recruited in this study. All of them had liquid-based thin-prep cytologic test (TCT), human papillomavirus (HPV) DNA testing and hTERC gene amplification analysis using interphase two-color FISH. In addition, colposcopically directed biopsy and/or cone biopsy were conducted for definite histopathologic diagnosis for each case. The optimal threashold of hTERC gene amplification by fluorescence in situ hybridization (FISH) were assecced by receiver operating characteristic (ROC) curve. The results of hTERC gene amplification analysis were compared with the cytological analysis, HPV DNA testing and those of subsequent biopsies. Results: Among the 109 patients, 18 were benign lesion, 17 were LSIL, 66 were HSIL and 8 were invasive carcinoma of cervix (ICC). Of them, hTERC-positive cases were found in 0.0% (0/18) of normal specimens, 11.8% (2/17) of LSIL, 72.7% (48/66) of HSIL and 100.0% (8/8) of ICC, respectively. The positive rate of hTERC gene amplification was significantly higher in HSIL and ICC compared with normal and LSIL (all P < 0.01).The optimal cut-off point of percentages of cells with hTERC amplification was determined as 5.5%. Using this threshold the hTERC test reached a much higher specificity(94.3%, 33/35) and a relatively lower sensitivity(77.0%, 57/74) to distinguish benign lesion and LSIL from HSIL and ICC in comparison with HR-HPV test (51.4%;91.9%) and TCT (74.3%;81.1%). Area Under the Curve revealed that hTERC amplification test performed more accurately (area under the curve = 0.857) compared to HPV test (area under the curve = 0.717) and cytology(area under the curve = 0.777) to discriminate HSIL or higher from LSIL or lower. This study also found a significant positive correlation between positive hTERC gain and HR-HPV infection, abnormal cytological or histopathologic lesions (all P < 0.01) in patients with cervical diseases. Conclusion: hTERC amplification testing may be a promising adjunct to screen women for cervical precancer or cancer with high specificity and accuracy.

Cloning and screening of scarless healing-related gene(s) in rabbit skin

Background: Over the past years, scientists have been working on the mechanisms of the scarless healing. The remarkable phenotypic differences between fetal and adult healing may lead us to find out their characteristics in genetics, which represent potentially important mechanisms to explain the differences in the quality of wound repair observed in fetus versus adult tissues. Methods: Middle laparotomy and hysterotomy were performed on pregnant rabbits on 20-day gestation to expose the fetal back, and longitudinal incision which penetrated full skin was made on the back of fetus. The trauma fetus skin was harvested at 12 h post-operation (FT), the fetus control (FC) and trauma adult skin (AT) were taken at the same time. dscDNA was synthesized from total RNA of skin samples with SMART technology. An improved suppression subtractive hybridization (SSH) method was applied to analyze the samples. Having taken one of the three samples as Tester respectively, the other two together as Drivers, one forward and two reverse hybridization products were gotten. Having amplified by selective PCR, the products were inserted into vector, and then transferred into E.coli HB101. The colonies were screened by electrophoresis, reverse Northern afterwards, and the positive clones were sequenced. BLAST in NCBI was performed to compare and analyze the positive clones (expressed sequence Tag, ESTs). Results: Totally 298 clones were gotten and 61 positive clones were obtained after screening. The 61 selected positive clones were sequenced and 54 sequences were goten. Conclusion: Instead of traditional SSH, an improved SSH with 2 Drivers was applied in the experiment. The improved program is reasonable and correct in both theory and practice.

Screening of Genes with Unique Mutations of Microcus

Yersinia pestis is the causative agent of bubonic and pneumonic plagues. Strains of Y. pestis are classified into four biovars： antiqua, mediaevalis, orientalis, and microtus[11. There are two microtus-related plague loci in China： the Microtus brandti plague focus in the Xilin Gol Grassland （focus L） and the Microtus fuscus plague focus in the Ojnghai-Tibet Plateau （focus M）.

Screening Mouse Genes Related to Blastocyst Implantation by Using PCR Subtraction

Objective To identify genes that may be related to embryo implantation Materials&Methods The PCR subtraction technique was applied at implantation and inter-plantation sites on day 4.5 of pregnancy in mice.Two novel Expressed Sequence Tags(ESTs),EST8 and EST81 were identified;their expression in tissues was analyzed by Northern blotting,and their full-length cDNAs were synthesized by PCR.Results We found that these two novel ESTs(EST8and EST81)were noticeably expressed in implantation site in the mouse on day 4.5 of pregnancy.EST8 was expressed at high level in livers and implantation sites of the mice,while at low level in ovaries and inter-plantation sites.EST81 was predominantly expressed in implantation site and ovary,and at low level in all other tissues.Their complete cDNAs,1665bp and 1264 bp respectively,were synthesized by using PCR.Conclusion The two full-length cDNAs were responsible for embryo implantation,and their functions need to be further studied.

Application of CRISPR-Cas9 technology in the screening of tumor resistance genes:A review

Drug resistance is a major problem faced by tumor cell-targeted drugs.Currently,functional gene screening is the most common strategy for screening drug resistance genes.In recent years,Crispr-cas9 gene editing technology has been widely used in the functional studies of tumor-related genes due to their characteristics of accuracy,simplicity and efficiency.The principle of CRISPR-Cas9 Library Screening Technology and its application in functional Gene Screening are reviewed.At the same time,the application prospect of the Crispr-Cas9 technology is forecasted.

Utilization of Gene Mapping and Candidate Gene Mutation Screening for Diagnosing Clinically Equivocal Conditions: A Norrie Disease Case Study

Prenatal diagnosis was requested for an undiagnosed eye disease showing X-linked inheritance in a family. No medical records existed for the affected family members..Mapping of the X chromosome and candidate gene mutation screening identified a c.C267A[p.F89L] mutation in NPD previously described as possibly causing Norrie disease..The detection of the c.C267A[p.F89L] variant in another unrelated family confirms the pathogenic nature of the mutation for the Norrie disease phenotype. Gene mapping, haplotype analysis, and candidate gene screening have been previously utilized in research applications but were applied here in a diagnostic setting due to the scarcity of available clinical information..The clinical diagnosis and mutation identification were critical for providing proper genetic counseling and prenatal diagnosis for this family.

Screening and Primary Characterization of NewAntigen Genes of Schistosoma Japonicum

To find Schistosoma japonicum(S.j)new antigen gene thus provide more useful vaccine candidates, the cDNA library of S.j adult worm was screened with sera of rabbits immunized with the membrane antigens of Schistosoma japonicum hepato-portal schistosomula (SjHmAg). The positive clones were amplified by PCR and sequenced, then the sequences of clones were compared with all sequences in GenBank database using Blast process. The new clones were submitted to GenBank for accession numbers. Fifteen positive clones were obtained after three rounds of immunoscreening. The size of S.j cDNA fragments in positive clones ranged from 0.7?kb-3.0?kb after automatically excised with the helper phage. Sequence analysis revealed that partial sequence of clone M5 had significant homology with S.j mitochondria mRNA, the other positive clones were new S.j genes. M2 clone sequence (GenBank accession number AF502579) was 730?bp long it had a 117?bp open reading frame (ORF). The sequence of M15 (GenBank accession number AF502582) has no transmembrane region and encodes 92 amino acids, and its protein contains a ferredoxins iron-sulfur binding region signature and two VWFC signal regions. The size of M1、M8、M9、M12(GenBank accession numbers: AF502578, AF502580, AF500622, AF502581) ranges from 402?bp to 766?bp. It concluded that the sera from rabbit immunized with SjHmAg could recognize S.j specific antigens molecules, and these antigens may induce the protective immunity against S.j infection.

A screening analysis of the GJB2 c.176 del 16 mutation responsible for hereditary deafness in a Chinese family

Objective:To determine whether a new-born child from a family carrying a deafness gene needs cochlear implantation to avoid dysphonia by screening and sequencing a deafness-related gene.Results:Both screening and sequencing results confirmed that the new born child had a normal GJB2 gene despite the fact that she has a brother suffering from hearing loss triggered by an allelic GJB2 c.176 del 16 mutation.We cloned the GJB2 genes derived from their respective blood genomic DNA into GFP fused plasmids and transfected those plasmids into the 293 T cell line to test for gene function.While the mutated GJB2gene(GJB2 c.176 del 16) of her deaf brother was found to be unable to form the gap junction structure between two adjacent cells,the baby girl’s GJB2 gene ran into no such problems.Conclusion:The screening and sequencing as well as the GJB2 gene function tests invariably showed results consistent with the ABR tested hearing phenotype,which means that the child,with a normal wild type GJB2 gene,does not need early intervention to prevent her from developing hearing loss and dysphonia at a later stage in life.

THE CLONING OF HRNT-1 USING A COMBINATION OF cDNA LIBRARY SCREENING WITH BIOTIN-LABELED PROBE AND RAPID AMPLIFICATION OF cDNA ENDS

Objective: To clone the human counterpart of rat ZA73, EST cloned from rat tracheal epithelial (RTE) neoplastic transformed cell model induced by (a-particles radiation by using mRNA differential display. Methods: According to the sequence of rat ZA73, a probe was biotin-labeled to screen human cDNA library, and then the gene sequence was extended by RACE (rapid amplification of cDNA ends). Result: Human gene HRNT-1 (GenBank Accession Number: AF223393) is 4.256 kb in length, with an ORF located in the region between 254 and 3013 bp. 5′ UTS (untranslated sequences) is 253 bp, 3′ UTS is 1243 bp. Conclusion: The combination of cDNA library screening with biotin-labeled probes and RACE is an effective method to clone full-length cDNA, especially for sequences longer than 2 kb.

Research progress and application of the CRISPR/Cas9 gene-editing technology based on hepatocellular carcinoma

Hepatocellular carcinoma(HCC)is now a common cause of cancer death,with no obvious change in patient survival over the past few years.Although the traditional therapeutic modalities for HCC patients mainly involved in surgery,chemotherapy,and radiotherapy,which have achieved admirable achievements,challenges are still existed,such as drug resistance and toxicity.The emerging gene therapy of clustered regularly interspaced short palindromic repeat/CRISPR-associated nuclease 9-based(CRISPR/Cas9),as an alternative to traditional treatment methods,has attracted considerable attention for eradicating resistant malignant tumors and regulating multiple crucial events of target gene-editing.Recently,advances in CRISPR/Cas9-based anti-drugs are presented at the intersection of science,such as chemistry,materials science,tumor biology,and genetics.In this review,the principle as well as statues of CRISPR/Cas9 technique were introduced first to show its feasibility.Additionally,the emphasis was placed on the applications of CRISPR/Cas9 technology in therapeutic HCC.Further,a broad overview of non-viral delivery systems for the CRISPR/Cas9-based anti-drugs in HCC treatment was summarized to delineate their design,action mechanisms,and anticancer applications.Finally,the limitations and prospects of current studies were also discussed,and we hope to provide comprehensively theoretical basis for the designing of anti-drugs.

Screening and molecular classification of low-temperature protease from Antarctic microorganism and its characterization

107 strains producing protease were screened from 260 strains of Antarctic psychrophilic bacteria, among which proteolytic activity of five strains was more than 45 U ml^-1. The 16S rRNA gcne sequences homology and phylogcnetic analysis of five Antarctic psychrophillc bacteria showed that NJ276, NJS-9, NJ16-70,NJ345 belonged tO the described genus Pseudoalteromonas and NJ341 belonged to the genus Colwellia. The growth and the protease characteristic of four Antarctic psychrophilic bacteria had been studied, and the result showed that the 6ptimal temperature for growth and protease-produeing of four strains was about 10℃. Their growth and protease-produeing were still high during incubatlng 2-5 days. The maximum proteolytic activity occurred at pH 9 for four Antarctic psychrophilic bacteria. The optimal temperature of protease action of both strains NJ276 and NJ5-9 was about 50℃, however, the optimal temperature of protease aetlon of both strains NJ341 and NJ345 was about 40 ℃, and their proteolytic activity under 0℃ exhibited nearly 30% of the maximum activity, but their thermal stabilities were weaker. These results indicated that proteases from NJ341 and NJ345 were low-temperature proteases.

A living cell-based fluorescent reporter for high-throughput screening of anti-tumor drugs

Suppression of cellular O-linkedβ-N-acetylglucosaminylation(O-Glc NAcylation)can repress proliferation and migration of various cancer cells,which opens a new avenue for cancer therapy.Based on the regulation of insulin gene transcription,we designed a cell-based fluorescent reporter capable of sensing cellular O-Glc NAcylation in HEK293 T cells.The fluorescent reporter mainly consists of a reporter(green fluorescent protein(GFP)),an internal reference(red fluorescent protein),and an operator(neuronal differentiation 1),which serves as a"sweet switch"to control GFP expression in response to cellular OGlc NAcylation changes.The fluorescent reporter can efficiently sense reduced levels of cellular OGlc NAcylation in several cell lines.Using the fluorescent reporter,we screened 120 natural products and obtained one compound,sesamin,which could markedly inhibit protein O-Glc NAcylation in He La and human colorectal carcinoma-116 cells and repress their migration in vitro.Altogether,the present study demonstrated the development of a novel strategy for anti-tumor drug screening,as well as for conducting gene transcription studies.

Screening strategy for gastrointestinal and hepatopancreatobiliary cancers in cystic fibrosis

Based on systematic review and meta-analysis,the risk for developing cancers in patients with cystic fibrosis(CF)is known to be significantly greater than in the general population,including site-specific cancers of the esophagus,small bowel,colon,liver,biliary tract,and pancreas.An even higher risk has been found in patients who have severe CF transmembrane conductance regulator(CFTR)genotypes or who have undergone organ transplantation and are immunosuppressed.The risk continues to rise as life expectancies steadily climb due to advancements in medical care and treatment for CF.The colorectal cancer risk is at such a high level that CF has now been declared a hereditary colon cancer syndrome by the Cystic Fibrosis Foundation.The CFTR gene has been stronglyassociated with the development of gastrointestinal(GI)cancers and mortality in the CF population.Even CF carriers have shown an increased rate of GI cancers compared to the general population.Several limitations exist with the reported guidelines for screening of GI and hepatopancreatobiliary cancers in the CF population,which are largely universal and are still emerging.There is a need for more precise screening based on specific risk factors,including CFTR mutation,medical co-morbidities(such as gastroesophageal reflux disease,distal intestinal obstruction syndrome,and diabetes mellitus),familial risks for each cancer,gender,age,and other factors.In this review,we propose changes to the guidelines for GI screening of patients with CF.With the development of CFTR modulators,additional studies are necessary to elucidate if there is an effect on cancer risk.

Incidence of Sickle Cell Disease and Other Hemoglobinopathies in Burkina Faso: Results of a Five-Year Systematic Neonatal Screening (2015-2019) in Four Urban Hospitals

Hemoglobinopathies, mainly Sickle cell disease (SCD), are the most common monogenic disorders in Africa. In Burkina Faso, data on these diseases are scarce, mainly hospital-based in Ouagadougou and its surroundings. In order to assess the incidence and allelic frequencies of the main hemoglobinopathies in newborns in Burkina Faso, we conducted a cross-sectional study from 2015 to 2019 in four hospitals. The study included babies of both sexes, regardless of ethnic group and parents’ hemoglobin status. It was a newborn screening and hemoglobin variants were detected using isoelectric focusing on cord blood samples and confirmed using hemoglobin electrophoresis by high-performance liquid chromatography. The proportions and cumulative incidences of the different hemoglobinopathies were computed. Hardy-Weinberg equilibrium law was applied to calculate genotypic and allelic frequencies. The significant level was p < 0.05. Out of 11,337 newborns included, 47.8% were males and 60.2% were from Bobo-Dioulasso. Abnormal hemoglobin was found in 27.1%, representing a cumulative incidence of 1:4 newborns. The incidence of SCD was 1.9% (1:53 newborns) with 27.9% of homozygous SS. Homozygous CC and compound heterozygous Cβ-Thalassemia accounted for 1.1%. SCD cases were 1.51 times higher in Bobo-Dioulasso (OR = 1.51;95% CI [1.09 - 2.10]: p = 0.013). The observed genotype frequencies were significantly different from the expected ones (p 0.001). The βS and βC alleles represented 5.1 and 9.9%, respectively. This study showed a high incidence of hemoglobinopathies. Such results raise the question of control strategies for these hemoglobinopathies in our country.

川东北地区3089例育龄期人群地中海贫血基因筛查结果分析

目的通过分析地中海贫血基因筛查结果,了解川东北地区育龄期人群中地中海贫血基因携带情况,为本地区地中海贫血的一级和二级防控提供实验室依据。方法回顾性分析2023年度在川北医学院附属医院进行地中海贫血基因筛查的3089例育龄期男女的基因检测、血常规和血红蛋白电泳结果,对地贫基因检测阴性而后两种方法检测结果异常者行罕见地中海贫血基因测序;选取其中22例血常规和血红蛋白电泳结果正常而本院基因检测结果提示为-ɑ3.7杂合缺失的标本进行HKɑɑ及ɑ三联体检测。结果3089标本中共检出370例地中海贫血基因携带者,阳性率为11.98%(370/3089),ɑ-地贫基因携带者194例(52.43%),β-地贫基因携带者166例(44.86%),最常见的基因型为--^(SEA)/ɑɑ、-ɑ^(3.7)/ɑɑ、β^(CD41-42M)/β^(N)、β^(CD17M)/β^(N)和β^(IVS-II-654M)/β^(N);罕见地贫基因测序标本中有1例为ɑɑɑ^(anti4.2),1例为β^(IVS-II-654M)杂合变异,4例为δ-地贫,1例为δ-地贫复合nd-HPFH,3例为结构异常血红蛋白,1例为HBB基因良性变异;PCR结果提示22例待测标本中有4例HKɑɑ。结论本地区育龄人群中的地中海贫血基因携带率较高,基因型复杂,应重视罕见地中海贫血基因和HKɑɑ的检测。

基于重症支气管哮喘差异表达基因及其治疗中药筛选的生物信息学分析

目的:通过生物信息学方法探讨重症支气管哮喘[简称重症哮喘(SA)]的差异表达基因,分析其作用机制,并筛选潜在具有治疗作用的中药及活性成分。方法:在高通量基因表达(GEO)数据库中选取GSE136587和GSE158752数据集,利用R软件对数据集进行差异分析获得差异表达基因,并进行蛋白-蛋白相互作用(PPI)网络分析,筛选核心基因,寻找关键通路和枢纽基因。最后将核心基因提交至Coremine数据库筛选具有潜在治疗作用的中药,并通过《中华医典》检索相关中药方剂。结果:共筛选出466个差异表达基因。通过STRING平台构建PPI网络共筛选包括25 kDa突触关联蛋白(SNAP25)、谷氨酸离子型受体2(GRIA2)、轴突蛋白1(NRXN1)、钾电压门控通道亚家族A成员1(KCNA1)、突触囊泡蛋白1(SYT1)和嗜铬蛋白A(CHGA)等核心靶点25个。基因本体(GO)功能富集显示SA的生物学过程与细胞趋化性和白细胞迁移等有重要关系,京都基因与基因组百科全书(KEGG)富集的通路主要涉及骨髓白细胞迁移、白细胞趋化性、细胞趋化性、白细胞迁移、对外部刺激反应的正向调节和骨髓白细胞活化等信号通路。采用网络药理学方法基于核心靶点筛选得到具有潜在治疗SA作用的中药367种,其中人参、水牛角、全蝎和黄芪等中药涉及多个核心靶点,与SA具有高度相关性,在《中华医典》中检索具有高度相关性的中药,共得到17个潜在具有治疗效果的中药方剂。结论:通过生物信息学筛选SA的潜在标志物和具有治疗作用的中药,为SA早期诊断和发病机制研究提供新的靶点,为其治疗的中药方剂研发提供思路。