Research Progress on Pathogenic Factors of Breast Cancer Combined with Thyroid Cancer / 肿瘤防治研究

Thyroid cancer and breast cancer are two of the most common malignant tumors in women. Concurrent tumors of the thyroid and breast are relatively rare in clinical practice; however, the incidence of such dual malignancies has recently increased. Researches conducted in the past mainly focused on the possible increase in the incidence of contralateral breast cancer, while the increased risk of synchronized thyroid cancer in women with breast cancer has attracted widespread attention recently. The specific mechanism has not been fully understood. This article reviews the pathogenic factors between these two diseases, and evaluates the etiological role of these factors in these double primary cancers, so as to provide a better basis for clinical practice.

Analysis of the nutritional status of 25-hydroxy Vitamin D in 7215 pregnant women and health education in the maternal and child health care hospital of Dalian / 中国实用护理杂志

Objective:To study the level and deficiency of 25-hydroxy vitamin D [25-(OH)D] in 7215 pregnant women in Dalian district, and provide a scientific evidence for the prevention of vitamin D deficiency and reasonable supplementation.Methods:During January to December of 2018, the basic information and blood samples of 7215 pregnant were collected in Maternal and Child Health Care hospital of Dalian. The level of serum 25-hydroxy vitamin D was detected by chemiluminescence method. All data was analyzed by SPSS 17.0 software.Results:In Dalian district, the average level of serum 25-(OH)D in 7215 pregnant women was 22.74(16.02, 28.62) ng/ml, and the deficiency rate of vitamin D was 33.89%, and the severe deficiency rate was 6.68%. The average level of serum 25-(OH)D in pregnant women aged 18-24 years old, 25-34 years old and 35-45 years old were 20.22(13.61, 25.57) ng/ml, 22.75(16.12, 28.42) ng/ml and 23.60(16.76, 29.92)ng/ml, respectively. With the increasing of pregnant age, the level and the deficiency rate of serum 25-(OH)D was gradually rising with significant difference( P<0.01), and the highest deficiency rate was pregnant aged 18~24 years old with 42.40%. Conclusion:The level of vitamin D of pregnant women in Dalian district was insufficient status, especially for young pregnant women. Health education for vitamin D deficiency should be focused on young pregnant women.

Echocardiographic features and outcome of restrictive foramen ovale in fetuses without cardiac malformations / 中华超声影像学杂志

Objective To investigate the clinical significance of restrictive foramen ovale ( RFO ) monitored by fetal echocardiography during the middle to late stage of pregnancy . Methods The detection rate ,echocardiographic features and outcome in fetuses with RFO without cardiac malformations from 7319 pregnant women received prenatal echocardiography were retrospectively reviewed and analyzed . Results RFO was found in 40 of 7319 (0 .55％ ) fetuses . The inclusion criteria including a narrow right to left shunt of less than 2 .5 mm in diameter across atrial septum , enlarged right atrium , increased right-to-left ventricular size ratio ,and increased size ratio of main pulmonary artery to aorta were present in 40 fetuses . The direct ultrasound characters of RFO included limited opening of oval valve ( 70％ ) and foramen ovale diameter less than 2 .5 mm (30％ ) . And atrial septal aneurysm ( 62 .5％ ) ,redundant primum atrial septum (57 .5％ ) ,abnormal ductus arteriosus ( 57 .5％ ) might also be present commonly in RFO . As the gestational weeks increased , the size ratio of right-to-left atrium , right-to-left ventricle and the main pulmonary artery to aorta also increased significantly( P =0 .004 , P <0 .001 , P <0 .001) . Among the 40 fetuses with RFO ,21 cases ( 52 .5％ ) gave birth in full term ,8 cases ( 20％ ) which were detected severe tricuspid regurgitation gave birth in early cesarean section ,5 cases ( 12 .5％ ) had induced labor and 6 cases (15％ ) were lost in the follow-up . Of the 29 newborns ,only 1 case died of heart failure ,and the other 28 subjects recovered both from heart structure or cardiac function within four months . Conclusions RFOwithout cardiac malformations presents echocardiographic features characterized by a narrow right to left shunt of less than 2 .5 mm in diameter across atrial septum . Fetal echocardiography can monitor the dynamic change of fetal heart structure and function based on the increase of right heart load and decrease of left heart volume ,which has important clinical significance for assessing fetal intrauterine condition and prognosis .

Two new lignan glucosides from caulis of Tinospora sinensis / 中草药

Objective To study the chemical constituents and anti-neuroinflammatory activity of the caulis of Tinospora sinensis as a "Yao" medicine. Methods The chemical constituents were isolated and purified by silica gel, Sephadex LH-20, ODS column chromatographies, and semi-preparative HPLC. The structures of these compounds were elucidated by extensive spectroscopic analyses and chemical methods. All compounds were evaluated for their anti-neuroinflammatory effect by inhibiting the nitric oxide (NO) production in lipopolysaccharide (LPS)-activated murine BV-2 microglial cells except for compounds 7 and 8. Results Thirteen compounds were isolated from the caulis of T. sinensis. They were identified as tinosposide C (1), tinosposide D (2), seco-isolariciresinol 9-O-β-glucopyranoside (3), (+)-pinoresinol 4-O-β-D-glucopyranoside (4), (+)-syringaresinol (5), tanegoside A (6), (E)-3-[(2,3-trans)-2-(4-hydroxy-3-methoxyphenyl)-3-hydroxymethyl-2,3-dihydrobenzo [b] [1,4]dioxin-6-yl]-N-(4-hydroxyphenethyl) acrylamide (7), thoreliamide B (8), trans-N-p-coumaroyltyramine (9), N-trans-feruloyltyramine (10), N-trans-caffeoyltyramine (11), grossamide K (12), and cis-grossamide K (13), seven of which exhibited significant anti-neuroinflammatory activity with IC50 values ranging from 1.46 to 51.25 μmol/L. Conclusion Compounds 1 and 2 are new compounds and their absolute configurations are confirmed by ECD experiments, compounds 3, 5, 7, 8, and 10-13 are isolated from the plants for the first time. The activity of compounds 9 and 10 is better than positive control minocycline.

Application of Exosomes Derived from Mesenchymal Stem Cells in Tissue Regeneration / 中国医科大学学报

Tissue regeneration involves the use of new tissue to replace or repair necrotic,damaged,or aged tissue. Exosomes,a type of extracellular vesicles secreted by cells,are important for the paracrine activity of stem cells and are involved in tissue regeneration. Recently, tissue regeneration strategies utilizing exosomes have gained considerable research attention. This review discusses the current advances in the application of exosomes derived from mesenchymal stem cells in tissue regeneration.

Double-bundle versus single-bundle anterior cruciate ligament reconstruction with allogeneic tendon via single femoral tunnel / 中国组织工程研究

BACKGROUND: The anterior cruciate ligament (ACL) is an extremely important static structure that stabilizes the structure of the knee joint. Many scholars have found that an insufficient understanding of normal anatomical structure and biological characteristics of the ACL results in poor outcomes in the ACL reconstruction. OBJECTIVE: To compare the clinical effects of single-bundle and double-bundle single-tunnel techniques in ACL reconstruction with allogeneic tendon. METHODS: (1) Anatomy research: Six fresh knee joints were used for the experiments, and X-ray, CT and MRI were taken for each sample to confirm no osteoclasia, tumor and collapse of ACL. Then the ACL was stripped from the tibial bone and used to run hematoxylin-eosin staining and observe the fiber distribution of each piece of the ACL. (2) Clinical research: A total of 40 cases of ACL arthroscopic reconstruction were involved in this study, including 19 single-bundle ones and 21 double-bundle ones. The International Knee Documentation Committee 2000 subjective knee form (IKDC2000), Tegner and Lysholm evaluations were performed in all the patients. Paired t-test was used to analyze the IKDC, Tegner and Lysholm scores before operation and at follow-up time. RESULTS AND CONCLUSION: Anatomy research: From the view of anatomy, there was no evidence to support the double beams of the ACL at the tibial end by hematoxylin-eosin staining. Clinical research: Forty patients (40 knee joints) were successfully followed for over 12 months, and their joint range of motion was normal. The IKDC, Tegner and Lysholm scores were all improved in both two groups after the surgery, while the Larson scores were significantly higher in the double-bundle group than the single-bundle group. To conclude, the single-tunnel double-bundle ACL reconstruction is better to restore the knee stability than the single-bundle ACL reconstruction. This is an easy operation that has the merits of operating conveniently, restoring the normal anatomy of the tibial end, and exhibiting excellent short-term therapeutic effects. However, its long-term effects are uncertain.

Effects of ursolic acid at different concentrations on proliferation and differentiation of osteoclasts and their significances / 吉林大学学报(医学版)

Objective:To investigate the effects of ursolic acid (UA) on the proliferation and differentiation of osteoclasts (OC), and to explore its role in orthodontic force-induced root resorption and its relationship with OC.Methods:The mononuclear / macrophage cells RAW264.7 were induced to the OC.Tacrolimus acid phosphatase (TRAP) staining and bone resorption lacunae observation were used to identify the induction.CCK-8 method was used to select the appropriate concentration of UA for RAW264.7 cell-free biotoxicity and to observe its effect on the proliferation and differentiation of RAW264.7.In experimental groups, UA with gradient concentrations (1.0,2.5,5.0,10.0,20.0and 40.0 μmol·L-1)were added.UA was not added in control group.Results:The TRAP staining and bone resorption lacunae observation showed that after the RAW264.7 cells were induced for 5 d, the TRAP staining positive cells were found;the resorption lacunae were rounded,and oval, etc,the bottom wall was coarser,and the boundary was clear,which indicated that the RAW264.7 cells were successfully differentiated into the osteoclasts.The CCK-8 detection results showed that high concentration of UA (> 10.0 μmol·L-1) significantly inhibited the proliferation of OC;the appropriate concentration of UA (5.0 μmol·L-1) was in the biological safety concentration range and could inhibit the OC proliferation;low concentration of UA (<2.5 μmol·L-1) had no effect.Conclusion:RANKL can induce the differentiation and maturation of RAW264.7 cells.UA is correlated with the proliferation and differentiation of OC;UA has inhibitory effect on OC at the appropriate concentration (5.0 μmol·L-1) in a time-dependent manner.

Upregulation and biological function of transmembrane protein 119 in osteosarcoma

Osteosarcoma is suggested to be caused by genetic and molecular alterations that disrupt osteoblast differentiation. Recent studies have reported that transmembrane protein 119 (TMEM119) contributes to osteoblast differentiation and bone development. However, the level of TMEM119 expression and its roles in osteosarcoma have not yet been elucidated. In the present study, TMEM119 mRNA and protein expression was found to be up-regulated in osteosarcoma compared with normal bone cyst tissues. The level of TMEM119 protein expression was strongly associated with tumor size, clinical stage, distant metastasis and overall survival time. Moreover, gene set enrichment analysis (GSEA) of the Gene Expression Omnibus (GEO) GSE42352 dataset revealed TMEM119 expression in osteosarcoma tissues to be positively correlated with cell cycle, apoptosis, metastasis and TGF-β signaling. We then knocked down TMEM119 expression in U2OS and MG63 cells using small interfering RNA, which revealed that downregulation of TMEM119 could inhibit the proliferation of osteosarcoma cells by inducing cell cycle arrest in G0/G1 phase and apoptosis. We also found that TMEM119 knockdown significantly inhibited cell migration and invasion, and decreased the expression of TGF-β pathway-related factors (BMP2, BMP7 and TGF-β). TGF-β application rescued the inhibitory effects of TMEM119 knockdown on osteosarcoma cell migration and invasion. Further in vitro experiments with a TGF-β inhibitor (SB431542) or BMP inhibitor (dorsomorphin) suggested that TMEM119 significantly promotes cell migration and invasion, partly through TGF-β/BMP signaling. In conclusion, our data support the notion that TMEM119 contributes to the proliferation, migration and invasion of osteosarcoma cells, and functions as an oncogene in osteosarcoma.

Screening of small molecule inhibitors for PLK1 PBD and evaluation of antitumor activities / 药学学报

With the method of fluorescence polarization (FP), we screened small molecule inhibitors for PLK1 PBD to identify the lead compounds for antitumor drugs. FP led to the identification of a potent hit, F083-0063, whose inhibition rate was (99.7±0.4)% at 10 μg·mL-1. The IC50 was calculated to be 1.9±0.1 μmol·L-1 using Graphpad Prism 5. The effect of the compound on cells' multiplication was measured by MTT assay which showed that F083-0063 inhibited the proliferation of many tumor cell lines. Flow cytometry analysis indicated that the F083-0063 promoted cell apoptosis and induced cell G2/M arrest. Migration abilities of cells, evaluated using scratch test, increased significantly in the presence of F083-0063 with the mi-gration rate as low as (37.6±0.7)% at 20 μmol·L-1. Molecular linkage technique found F083-0063 had good affinity with PLK1 PBD. The results of Western blotting showed that the expression of cyclin-dependent proteins was increased after treatment with F083-0063. In summary, F083-0063 has an antitumor activity and is expected to be an antitumor lead compound targeting PLK1 PBD.

Ease effects of local inj ection of ursolic acid on orthodontic tooth movement and root resorption of rats and their mechanisms / 吉林大学学报(医学版)

Objective:To investgate the ease effects of local injection of ursolic acid both on orthodontic tooth movement distance and tooth root resorption in the rats, and to clarify its mechanism.Methods:Ninety-six male Wistar rats were used to establish models of orthodontic tooth movement.After the successful establishment of models,all the model rats were randomly divided into 0 (control group),0.5,1.0 and 2.0 mmol·L-1 ursolic acid groups (n=24).The rats were locally injected with different doses of ursolic acid into the palatal submucosal area adjacent to the right upper first molar ,once per 3 d,each time 50μL.The rats were respectively sacrificed on the days 1,3,5,7,10,14,21 and 28,then the distances of tooth movement were measured.HE staining was used to observe the morphologic changes of the root tissue of rats.Results:The tooth movement distances of rats in control,0.5,1.0,and 2.0 mmol· L-1 ursolic acid groups were increased with the prolongation of time (P0.05),but at the other time points,the tooth movement distances of rats had statistical significance (P<0.01).The tooth movement distances of rats had statistical significance between 0.5,1.0 and 2.0 mmol·L-1 ursolic acid groups 5,7,10,14,21 and 28 d after forcing (P<0.05 or P<0.01).The morphological results showed that the resorption pits were found on the surface of tooth root with the prolongation of loading time;in addition,with the increasing of the doses of ursolic acid, the absorption of root surface was relieved. Conclusion:Local injection of ursolic acid at the doses of 0.5,1.0 and 2.0 mmol·L-1 can reduce the distance of orthodontic tooth movement in the rats, and the distance of tooth movement in the rats is reduced with the increasing of its doses,and ursolic acid has the ease effect on the orthodoutic tooth root resorption.

The measurement of medical students' learning passion and the logistic regression analysis of influ-ence factors / 中华医学教育探索杂志

Objective To measure the learning passion of medical students and evaluating its in-fluencing factors. Methods Taking 879 medical students as research subjects to conduct a questionnaire survey according to specialty and grade stratified sampling. The questionnaire contained two parts, includ-ing learning passion scale and general situation questionnaire. The effective recovery rate was 69.28%, 609 valid questionnaires were recovered. SPSS 22.0 and AMOS 21.0 software were used for statistical analysis of data, analyzing the reliability and validity of the questionnaire with internal consistency reliability coeffi-cient and confirmatory factor analysis. The factors of learning passion of medical students were analyzed by logistic regression analysis. Results The questionnaire of learning passion of medical students contained 12 measurement items, including 2 dimensions: harmonious passion and obsessive passion. The questionnaire was of fine reliability (Cronbach's Alpha=0.916) and validity (χ2/df=3.401,RMSEA=0.073,GFI=0.958). The learning passion of medical students was at upper middle level (4.390±0.934). The influence of achievement level (OR=1.691, 95%CI=1.415 to 2.021), school satisfaction (OR=0.586, 95%CI=0.402 to 0.854) and profession plan (short-term plan OR=2.121, 95%CI=1.310 to 3.434;long-term plan OR=3.822,95%CI=1.972 to 7.405) on learning passion were statistically significant. Conclusion The questionnaire of learning pas-sion of medical students has fine reliability and validity. Achievement level, school satisfaction and profes-sion plan are factors affecting the learning passion of medical students.

Inhibition effect of miR-577 on hepatocellular carcinoma cell growth via targeting β-catenin

OBJECTIVE@#To investigate the expression and the regulation effect of cell growth of microRNA-577 in hepatocellular carcinoma (HCC).@*METHODS@#qRT-PCR was applied to detect the relative expression of miR-577 in 70 paired HCC and matched tumor adjacent tissues collecting from resection between March 2011 and March 2014. Pearson chi-square test was used to analyze the relationship between the miR-577 expression and clinical features. The miR-577 mimics were transfected into HepG2 cells; cell cycles were detected by flow cytometry, cell proliferation was measured by MTT assay and BrdU incorporation assay, and cell apoptosis was determined by flow cytometry and caspase3/7 activity analysis. The expressions of β-catenin were measured by immunohistochemistry. Spearman correlation analysis was used to analyze the relationship between miR-577 and β-catenin. qRT-PCR and western-blot were used to detect the expression of β-catenin in transfected HepG2 cells.@*RESULTS@#The relative expressions of miR-577 was significantly lower in HCC tissues compared to the matched normal tumor-adjacent tissues (P < 0.05). Low expression of miR-577 was significantly associated with large tumor size (≥5 cm, P < 0.05) and advanced tumor node metastasis stage (III+IV, P < 0.05). Transfection of miR-577 mimics could inhibit repress cell proliferation, enhance cell apoptosis and block the cell cycles in G0/G1 phase (P < 0.05). miR-577 in HCC group had a significant negative correlation relationship with the expression of downstream target of β-catenin (P < 0.05). Both the mRNA and protein expression in HepG2 cells were down-regulated after transfection (P < 0.05).@*CONCLUSIONS@#Low expression of miR-577 is related to the malignant clinicopathological features in HCC tissues, and miR-577 may suppress HCC growth through down-regulating β-catenin.

Inhibition effect of miR-577 on hepatocellular carcinoma cell growth via targeting β-catenin

Objective: To investigate the expression and the regulation effect of cell growth of microRNA-577 in hepatocellular carcinoma (HCC). Methods: qRT-PCR was applied to detect the relative expression of miR-577 in 70 paired HCC and matched tumor adjacent tissues collecting from resection between March 2011 and March 2014. Pearson chi-square test was used to analyze the relationship between the miR-577 expression and clinical features. The miR-577 mimics were transfected into HepG2 cells; cell cycles were detected by flow cytometry, cell proliferation was measured by MTT assay and BrdU incorporation assay, and cell apoptosis was determined by flow cytometry and caspase3/7 activity analysis. The expressions of β-catenin were measured by immunohistochemistry. Spearman correlation analysis was used to analyze the relationship between miR-577 and β-catenin. qRT-PCR and western-blot were used to detect the expression of β-catenin in transfected HepG2 cells. Results: The relative expressions of miR-577 was significantly lower in HCC tissues compared to the matched normal tumor-adjacent tissues (P < 0.05). Low expression of miR-577 was significantly associated with large tumor size (≥5 cm, P < 0.05) and advanced tumor node metastasis stage (III+IV, P < 0.05). Transfection of miR-577 mimics could inhibit repress cell proliferation, enhance cell apoptosis and block the cell cycles in G

Establishment and evaluation of method to collect rat gingival crevicular fluid by using absorbent paper points / 吉林大学学报(医学版)

Objective To establish and evaluate the method to collect the rat gingival crevicular fluid (GCF)by using absorbent paper points, and to lay foundation for analysis on GCF.Methods 20 healthy male rats were selected and randomly divided into GCF group and saliva group.The GCF of the right upper molar gingival trough of the rats in GCF group and the saliva of the rats in saliva group were collected by using 1 5# absorbent paper points.The SDS-PAGE analysis and abundance detection were applied to analyze the protein bands of the samples in two groups.Results The SDS-PAGE analysis identified the proteins at 77 000,66 000,55 000,51 000,and 28 000,especially 66 000 in GCF group.While saliva group had lower brightness protein bands at 66 000,60 000, and 48 000.The data of protein abundance of 66 000 between two groups had statistically significant difference (P<0.05).Conclusion The number and types of the protein bands are different between GCF Group and saliva group,so using 15# absorbent paper points can collect the rat GCF successfully.

Advance research on space analysis related factors / 医学研究生学报

Orthodontics is the procedure of managing the space .Space analysis is a comprehensive quantitative analysis that can analyze the factors that influence the arch space .It also helps to judge if the treatment goals can be reached , thus helping the treat-ment plan making and guiding clinical orthodontics .And space analysis related factors include crowding , curve of Spee, arch width, arch growth, arch protrusion, mesiodistal angulation, an so on.In this review, we summarizes the above-mentioned factors.

Advance research on protein tyrosine phosphatase SHP-2 and the regulating role of it in osteoclastogenesis / 医学研究生学报

SHP-2 is one of the protein tyrosine phosphatases which plays a role in the progress of dephosphorylation in organ -isms, participating in many kinds of signal transduction pathways .The mutation of SHP-2 is associated with many kinds of malignant diseases .In recent years , scholars have found that SHP-2 is the key factor in osteoclastogenesis , playing a positive role in the progress of reversible phosphorylation of osteoclasts .The objective of this article is to review the molecular biological characteristics of SHP-2, th diseases associated with the mutation of SHP-2 and the effects of SHP-2 in regulation of osteoclastogenesis .

Nursing of patients with nasal facial soft tissue defect treated with transplantation of free foot skin flap / 现代临床护理

ObjectiveTo summarize the nursing measures for patients with nasal facial soft tissue defect treated with transplantation of free foot skin flap.MethodThe clinical data of 13 patients with transplantation were reviewed to summarize the nursing measures.ResultAll skin flaps of the 13 patients survived.ConclusionEnough preoperative preparation,active psychological nursing and careful postoperative observation are critical for the success of the operation.

Pain experience during initial alignment with self-ligating and conventional brackets

The aim of this study was to compare the pain experience among orthodontic patients treated with self-ligating brackets SmartClip® (3M Unitek, Monrovia, California, USA) and conventional brackets Victory series® (3M Unitek, Monrovia, California, USA). We used a controlled clinical trial study design to compare 69 patients treated with self-ligating to 70 patients treated by conventional brackets. The nickel-titanium archwires 0.012-in were engaged after bonding both arches on the first day; and the visual analogue scale (VAS) was used to assess the pain experience of subjects for the first seven treatment days. The pre-treatment dental study models were assessed by the Little’s irregularity index to quantify the groups’ malalignment characteristics. The self-ligating brackets reported lower pain experience than the conventional group on the first five treatment days. However, the sixth day showed 1.75 mm higher visual analogue score than conventional brackets, with almost equal pain level on the seventh day. The group differences throughout the first week were neither clinically nor statistically significant. The pain experiences in both groups decreased steadily from the third treatment day to the end of the first week of treatment. Based on the study findings, the pain experience during initial alignment is not influenced by the brackets’ ligation type. The pain experience tends to decrease steadily from the third treatment day to the end of the first week of treatment irrespective of the bracket type used.

Cost-effective production of protein by using cellulose-binding domain fusion tag in Corynebacterium glutamicum / 生物工程学报

The CBD gene from Trichoderma reesei was cloned into the Corynebacterium glutamicum secretion expression vector pXMJ19-sp, in which green fluorescent protein was inserted to obtain pXMJ19-sp-GFP-CBD. After induced by 0.5 mmol/L IPTG, GFP-CBD was expressed in Corynebacterium glutamicum at high level of 200 mg/L. The GFP-CBD could be purified to high purity with cellulose column. The results indicated CBD can be successfully used in Corynebacterium glutamicum expression system and thus offer an extremely simple, effective and scalable way for production of recombinant proteins.

Utility of high throughput sequencing technology in analyzing the terminal sequence of caudovirales bacteriophage genome / 病毒学报

To confirm the hypothesis that the high frequency sequences of high throughput sequencing are the terminal sequences of the bacteriophage genome. An adaptor of specific sequence was linked to the end of the bacteriophage T3 genomic DNA, which was then subject to high throughput sequencing; as a control, the same T3 genomic DNA without adaptor was also analyzed by high throughput sequencing. The sequencing results were examined with bioinformatics software. Similar high throughput sequencing technique was applied to analyze the genomic sequence of N4-like bacteriophage IME11. Bioinformatics study showed that the sequences tagged with adaptors were consistent with the high frequency sequences without adaptor labeling. Our analysis also indicated that the end of the T4-like phage genome had specific sequences instead of random sequences, disagreeing with the previous assertion. Evidences were provided that N4-like bacteriophage had a particular terminal sequence: the left end of the genome was unique while the right end was permuted. The high throughput sequencing technique was convenient and practical to be used to simultaneously detect the terminal sequence and the complete sequence of bacteriophage genome.