Cytogenetic characterization of olive flounder Paralichthys olivaceus: DNA content, karyotype, AgNORs and location of major ribosomal genes

A cytogenetic analysis of Paralichthys olivaceus was carried out using the flow cytometry method for DNA content, silver staining for the nucleolus organizer region （AgNORs） identification and one-color fluorescence in situ hybridization （FISH） for chromosomal mapping of major ribosomal genes. Nuclear DNA content was estimated by flow cytometry method using Gallus domesticus erythrocytes as the internal reference standard. The C-value of this species was （0.737±0.024） pg, and the DNA contents of each chromosome were estimated to be 16.51 Mb to 39.50 Mb after paired according to the average relative length. The FISH probe was made by PCR amplification of a DNA fragment containing internal transcribed spacers ITS1 between 18S and 5.8S ribosomal RNA gene, and labeled by PCR incorporation of bio-16-dUTP. FISH signals and AgNORs were both located on the secondary constrictions of chromosome 1. These results will provide a better understanding of the cytogenetic information of this species and would help for further research of the karyotype evolution in the order Pleuronectiformes.

Analysis of DNA Content of Various Types of Spermatogenic Cells in Rat after Testicular Heating with Flow Cytometry

To measure DNA contents of spermatogenic cells and analyze the efficiency of spermatogenesis after testicular heating in rat Methods Eighty adult male Sprague-Dawley rats were randomly divided into experimental group （43 ℃, 30 min） and control group （22 ℃, 30 min）. According to day 0.5, 1, 3, 6, 10, 25, 35 and 50 after local testicular heating, every group was divided into 8 subgroups： experimental subgroups （n=6） and control subgroups （n=4）. DNA contents of various types of germ cells were observed with flow cytometry （FCM） in all groups. Results Compared with control groups, percentages of 4C cell （primary spermatocyte） in 0.5-35 d groups and percentages of 1C cell （spermatid and sperm） in 6-50 d groups significantly decreased in experimental groups （P〈0.05）, and percentages of 2C cell （spermatogonium and second spermatocyte） in 3 -35 d experimental groups increased significantly after heating （P〈0.05）. 4C：2C in all of 8 experimental groups and 1C：2C in 3-35 d experimental groups were down （P〈0. 05）, and in 1 d experimental group 1C：4C was up after heating （P〈0.05）. Conclusions After being heated, the number of spermatocyte firstly decreased, and then that of spermatid and sperm decreased too. Heat influences several stages in spermatogenesis and results in suppression of spermatogenesis. Flow cytometry is an effective method for researching on the change of spermatogenesis and has significance on mechanism about changing of spermatogenic cells induced by heat.

DNA Content in Pancreatic ExocrinalCells after Vagotom yand Electron Microscopy in Rats

Fifty six SD rats were randomly divided into the normal control group and the operation group. The operating group was subdivided into six groups on the basis of killing time (the 12th h, the first day, 3rd day, first week, 2nd week and 4th week) after vagotomy (VG). The pancreatic tissues were taken for HE and Feulgen staining. The DNA content of pancreatic exocrine cells was determined by a domestically fabricated computer image analyzing system. In the control group, on the first day or in the first week after VG, the pancreatic samples were taken for transmission electron microscopic examination. The DNA content of pancreatic exocrinal cells was decreased from 1 to 3 days after VG. The secretion was found to be in inhibitory state and One week later, it gradually restored. The results indicated that the proliferation and the function of the SD rat's pancreatic exocrinal cells were prohibited at initial stage after VG, which might be concerned, at least in part, with dominance and nutrition of vagus.

EFFECTS OF PERIOPERATIVE CIMETIDINE ADMINISTRATION ON TUMOR CELL NUCLEAR MORPHOMETRY AND DNA CONTENT IN PATIENTS WITH GASTROINTESTINAL CANCER

Objective: To explore the effects of perioperative cimetidine administration on tumor cell nuclear morphometric parameters and DNA content in patients with gastrointestinal adenocarcinoma. Methods: 49 patients with pathologically confirmed gastrointestinal adenocarcinoma were randomized into test group (n=25) and control group (n=24). The test group started oral cimetidine intake 400 mg, tid, 7–10d before operation, followed by standard curative operation. The control group did not receive cimetidine. Tumor specimens were paraffin embedded for microsection and stained with hematoxylin and eosin (HE) and Feulgen stain. Morphometric studies and DNA content of tumor nuclei were performed on IBAS Image Analyzer. Results: The tumor cell nuclear area (μm2), nuclear perimeter (μm), maximal nuclear diameter (μ) for test group/control group were 23.54 5.08/34.69110.08 (P<0.001), 22.064.43/24.884.05 (P<0.05), 7.8411.64/ 8.6211.24 (P<0.05), 4.4210.61/5.4110.89 (P<0.001), Respectively. The percentages (%) of diploidy, triple-tetraploidy, quintuple ploidy, and >quintuple ploidy tumor cells for test group/control group were 16.6412.58/5.3312.14 (P<0.002), 39.8412.28/35.7013.58 (P>0.50), 12.4215.00/14.4810.74 (P>0.20), 31.1116.86/ 45.9713.82 (P<0.005), respectively. Conclusion: Perioperative administration of cimetidine in gasgtrointestinal cancer patients could decrease the nuclear size and raise the percentage of diploid tumor cells, and convert high aneuploid tumor cells into low-aneuploid tumor cells, which might help reduce the invasiveness of tumor cells.

Effectof A-L Tonic Capsule on DNA Contentin Rat Experimental Hepatocarcinogenesi

The effects of A L tonic capsule on DNA content in rat experimental hepatocarcinogenesis induced by diethylnitrosamine (DENA) were observed. The experimental rats were divided into 4 groups. With exception of group D in which the rats were only administered with DENA, the rats in the groups A, B, C were previously, simultaneously and subsequently fed with A L tonic capsule respectively while they were administered with DENA. The DNA content of all rat livers was measured using automatic image analysis technique 20 weeks after administration of DENA. The results showed that the highest and lowest DNA contents were respectively seen in the groups D and A. There was significant difference in DNA contents between the groups A or B or C and D, and also between the groups A and B or C (both P <0.01). 4 components (4C) and ≥5C cells were predominant in the group D, while 2C cells were the minority. The number of 2C cells in the groups A, B, C was significantly higher than that in the group D, and the number of ≥5C cells in the groups A, B, C was markedly lower than that in the group D ( P <0.01). Also, there was very significant difference in the number of 2C and ≥5C cells between group A and B or C ( P <0.01). It was concluded that A L tonic capsule could effectively inhibit the increase of DNA content of hepatocytes and improve the distribution of DNA content in rat hepatocarcinogenesis, especially in group A.

QUANTITATIVE STUDIES ON THE DNA CONTENT AND MORPHOMETRIC FEATURES OF LIVER CELL DYSPLASIA

The DNA content and morphometric features of hepatocellular carcinoma (HCC) and liver cell dysplasia (LCD), including nuclear area, nuclear perimeter, nuclear maximum diameter and nuclear circle diameter, were quantitatively determined by means of image analysis technology. The results showed that in comparison with normal hepatocytes, LCD had a markedly increased DNA content and nuclear morphometric parameters, but the values were lower than those for HCC. LCD showed a slight increase in nuclear atypia represented by the nuclear irregular index, which was also less than HCC. The findings indicate that LCD may be a precaneerous lesion of HCC, to the cells in an abnormal proliferative state.

DETERMINATION OF NUCLEAR DNA CONTENT IN LEIOMYOMA AND LEIOMYOSARCOMA OF GASTROINTESTINAL TRACT

Nuclear DNA content was measured by micro-spec trophotometry in 15 biopsy specimens from patients with gastrointestinal (GI) tract smooth muscle tumors (3 leiomyomas and 12 leiomyosarco-mas subdivided into 3 groups with 4 cases to each). The mean DNA value increased steadily as follows: leiomyoma (14.39±0.62 Au); leiomyosarcoma Grade Ⅰ (19.78±2.39 Au); leiomyosarcoma Grade Ⅱ (26.39± 1.60 Au); leiomyosarcoma Grade Ⅲ (30.66±2.39 Au). The difference of DNA value in the 4 groups had statistical significance (p<0.05-0.01). These results suggest that microspectrophotometric measurement of nuclear DNA content may serve as an objective quantitative method for the diagnosis of GI tract smooth muscle tumors and classification of leiomyosarcoma.

FLOW CYTOM ETRIC ANALYSIS OF CELLULAR DNA CONTENT IN EPITHELIAL OVARIAN TUMOR

The DNA content of tumor all was analyzed by flow cytometry on parafflnembedded specimens in 73 patients with epithelial ovarian tumor, and its clinical significance was evaluated. One of the 5 benign (20%), 2 of the 11 borderline (18.18%), and 30 of the 57 malignant (52. 63%) tumors were aneuplold. The occurrence rate of aneuploidy In malignant tumors was higher than In benign and borderline tumors ( P < 0. 05 ). Furthermore, aneuploidy was more frequently In the advanced stages (Ⅲ -Ⅳ ) (77. 7%) than in the early stages (Ⅰ - Ⅱ ) (9. 5%) (P<0. 005). The occurrence rate of DNA aneuploidy was higher in patients associated with ascites and the residual tumor≥.2 cm. Patients with aneuploid tumors had more of ten ascites (P<0. 005) and residual tumor size≥2cm (P< 0.005). There was no apparent correlation between the DNA ptoidy and the histologic grade, histologic type of the tumors. G0/G1 cell proportion of DNA diplold tumors in advanced carcinoma (64. 6%) was less than those of early stage carcinoma (75. 9% ) (P<0. 05). The survival rate of diplold tumor patients was higher than that of aneuploid tumor patients in the different time after operation, and the median survival time was 30. 2 months and 10. 3 months, respectively. Multivariate analysis revealed that cellular DNA ploidy was the most Important predictive factor (P = 0. 007) of prognosis, followed by residual tumor size (P= 0. 05). Different tumor specimen of the same patient can exhibit variation sometime (38. 9%).The results revealed that the DNA ploidy may reflect tumor biological characteristics, I. e. , Its proliferative ability. Analysis of cellular DNA content of epithelial ovarian tumors would help us to predict the prognosis of the patients better.

Conserved balance of hepatocyte nuclear DNA content in mononuclear and binuclear hepatocyte populations during the course of chronic viral hepatitis

瞄准：到有增加的原子 DNA 的 hepatocytes 的百分比满足的分析，即，四倍的(4n ) 和 octoploid (8n ) 原子核，然后比较单音原子并且双性人原子 hepatocyte 人口。方法：百分比单音原子双( 2n )，双性人的 4n ，和 8n hepatocytes 和那些原子 2 x 2n ， 2 x 4n ，并且 2 x 8n hepatocytes 与一个方法被决定能同时与长期的肝炎 B 或 C 在 62 个病人测量原子 DNA 满足的 hepatocyte 和 binuclearity 。在单音的原子 hepatocyte 人口的 4n 和 8n hepatocytes 的百分比在双性人与 2 x 4n 和 2 x 8n hepatocytes 的百分比相比原子 hepatocyte 人口。结果：在双性人的在单音的原子 hepatocytes 的 4n 和 2 x 4n 和 2 x 8n hepatocytes 的百分比原子 hepatocytes 是类似的，不管这项活动或长期的肝炎并且不管感染的病毒的纤维变性等级。结论：在内的原子 DNA 内容的分发单音原子并且双性人原子 hepatocyte 人口在长期的病毒的肝炎的功课期间被保存。

The Value of p21^(WAF1), p53 Protein and DNA Content in Large Intestine Cancer

目的 探讨 p2 1WAF1 蛋白和 p5 3蛋白在大肠癌发生发展中的作用及其临床意义。方法 对 4 1例大肠腺癌标本及 10例正常大肠粘膜标本 ,应用免疫组织化学S -P法检测细胞周期调控因子 p2 1WAF1 蛋白和突变型 p5 3蛋白 ,应用流式细胞术检测新鲜标本DNA含量。结果 p2 1WAF1 与 p5 3表达呈负相关 ;在正常大肠粘膜、高分化腺癌、中分化腺癌、低分化腺癌中 p5 3表达率逐渐增高 ,p2 1WAF1 表达率逐渐降低 ,SPF值逐渐升高 ;Dukes分期越晚 ,p2 1WAF1 表达率越低 ,p5 3表达率越高 ,SPF越高。结论 本实验证实了 p2 1WAF1 和 p5 3表达在肿瘤发生发展中起着调控点的作用 ;p2 1WAF1 、p5

硒对云南白灵芝生长、品质及DNA甲基化特征的影响

采用田间覆土栽培试验,设置6个Se浓度(0、50、100、200、300、400μg/g)处理,分别记为CK、Se50、Se100、Se200、Se300、Se400,研究外源硒对云南白灵芝生长发育、重金属积累、生物活性成分含量及其DNA甲基化特征的影响。结果表明,与CK相比,外源Se对子实体柄长和菌盖大小(横径、纵径)无显著影响,但能显著降低菌盖厚度;随着Se施用水平提高,菌丝生长速率和子实体硒含量均呈先增后降的趋势,生物量呈先降后增再降的趋势。子实体重金属含量(As、Cd、Pb、Hg)随Se浓度增加呈持续降低趋势,线性分析也表明,重金属含量与Se施用浓度均呈显著负相关。施Se增加了子实体多糖、粗蛋白、黄酮及总氨基酸含量,其中,多糖、黄酮含量在Se400处理下最高,总氨基酸、粗蛋白含量分别在Se200、Se300处理下最高。HPLC测定分析表明,与CK相比,Se300提高了灵芝酸A、灵芝酸DM、灵芝酸F及灵芝酮三醇为主的三萜酸组分,其总三萜酸含量较其他处理提高2.93%~25.54%。敏感扩增多态性分析(MSAP)结果显示,Se300较CK、Se400相比具有较高的未甲基化位点比例和较低的甲基化位点比例。综上,施用外源硒可促进早期菌丝生长,提高子实体生物量累积,改善子实体重金属安全性能和生物活性成分品质,改变DNA甲基化比例,且以300μg/g施用浓度的整体效果最佳,其子实体产量较对照显著提高11.36%。

脱落细胞DNA定量分析技术动态监测口腔扁平苔藓伴上皮异常增生病情及光动力治疗1例

口腔扁平苔藓是一种临床上常见的慢性炎性口腔黏膜病,具有癌变风险,临床上应积极干预并随访监测。光动力治疗具有微创、有效、选择性好以及可重复性高等优势,在治疗口腔扁平苔藓中已显现出良好疗效,但其疗效评估方法仍然存在局限性。脱落细胞DNA定量分析技术是一种无创辅助检查手段,可用于口腔潜在恶性疾患癌变及口腔癌的早筛早诊。本文报道1例采用脱落细胞DNA定量分析技术动态监测口腔扁平苔藓癌变风险及光动力治疗疗效的诊疗过程,以期为建立口腔扁平苔藓伴上皮异常增生的精准微创诊疗体系提供参考。

利用基于Python/RGB模块的DNA电泳图像分析方法检测绵羊血浆中羊源性成分

该文探究了运用Python处理食品中DNA分子量与含量测定的新方法,建立了DNA凝胶图像分析方法。将不同分子量的DNA Marker与DNA标准样品进行琼脂糖凝胶电泳并拍照,利用自行开发Python程序分析凝胶图,对图像进行灰度图转换、高斯模糊、图像阈值化、轮廓检测的图像优化步骤,探究了轮廓平均值法、轮廓中线法、全局数据平均法、全局数据积分法反映出DNA浓度与RGB数值间的线性关系,选取最优处理方法,通过读取像素迁移距离、RGB-灰度、RGB-向量、RGB-亮度进行DNA分子量与含量的测定实验,建立一种基于Python/RGB色彩体系的DNA凝胶电泳中分子量与含量的分析方法。检测结果误差较小,证明了Python/RGB的DNA分子量与含量分析方法的可行性,同时将该文所构建的凝胶图像分析方法应用于绵羊血浆中羊源性成分检测,结果显示所得目的蛋白为296 bp,而用DNA检测法得出样品中片段大小为294 bp,误差为0.99%,有望构建一种肉类源性成分检测的新方法。

高效液相色谱-示差折光法测定NMM抗肿瘤DNA疫苗中甘露醇含量

目的 建立测定NMM抗肿瘤DNA疫苗中甘露醇含量的高效液相色谱-示差折光法。方法 检测器为示差折光检测器,色谱柱为SUGAR SC1011阳离子钙型交换柱(300 mm×8 mm,6μm),流动相为纯化水,流速为1.0 mL/min,柱温为80℃,检测池温度为37℃,进样量为10μL。结果 甘露醇的质量浓度在0.02~5.00 mg/mL(R2=1.000 0,n=6)范围内与峰面积线性关系良好;定量限为13.37μg/mL;精密度、重复性、稳定性试验结果的RSD均小于2.0%;加样回收率为97.76%,RSD为0.97%(n=9)。3批小试样品和中试样品中甘露醇的含量分别为20.02,19.95 mg/mL。结论 该方法操作简便、重复性好、结果准确,可用于NMM抗肿瘤DNA疫苗中甘露醇的含量测定。

乙肝大小三阳状态、不同HBVDNA载量孕妇肝功能指标水平分析

目的研究探讨乙肝孕妇大三阳和小三阳状态中乙型肝炎病毒(Hepatitis B virus,HBV)DNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能水平。方法选取临清市人民医院于2020年1月—2023年1月收治的120例乙肝孕妇作为研究对象,根据乙肝两对半检测结果将其分为两组,即大三阳组(n=60)和小三阳组(n=60)。比较两组HBVDNA载量、肝功能指标、肝纤维化标志物含量及不同HBVDNA载量孕妇的肝功能指标、肝纤维化标志物含量。结果大三阳组丙氨酸转移酶水平(29.00±7.62)U/L、HBVDNA水平(5.18±1.16)×10^(4)U/mL均高于小三阳组的(26.00±2.85)U/L、(2.08±1.11)×10^(4)U/mL,差异有统计学意义(t=2.856、14.956,P均<0.05)。大三阳组的重组人几丁质酶3样蛋白1水平低于小三阳组,差异有统计学意义(P<0.05)。伴随着HBVDNA载量上升,两组患者甲胎蛋白水平随之增加,但是在同等HBADNA载量下,两组甲胎蛋白水平比较,差异无统计学意义(P>0.05)。结论开展孕前抗乙肝病毒诊疗、孕期风险系数评估,采取科学有效的干预措施,均能够有效抑制乙肝在母婴中的传播。

Flow Cytometry Analysis of Celullar DNA Content of the Parotid Pleomorphic Adenoma

The nuclear DNA content in 42 cases of primary parotid pleomorphic adenoma (PPA) and 15 cases of recurrent PPA were analysed with flowcytometer (FCM) to study the correlation of the DNA index (DI), S% and cellular proliferous index (PI) with the biologic behaivor of the tumor. The results indicated that in a portion of pleomorphic adenoma, the nuclear DNA content had altered and possessed malignant potential before any evidence of malignancy could be found under lightmicroscope. The increase of S% and PI, which followed the course of tumor development, has a close relation with the capsular invasion and the recurrence. There was a significant difference on the DI, S%, PI as well as the incidence of heteroploid between the recurrent and the incipient tumor. A great portion of recurrent tumors, which still diagnosed as benign pathologically, was acually heteroploid.

CHANGES OF CELLULAR MORPHOLOGY AND NUCLEAR DNA CONTENT IN DIFFERENT NASOPHARYNGEAL EPITHELIUM FROM DIFFERENT PATIENTS

Cellular morphology and nuclear DNA content were measured with point-counting technique and flow cytometry (FCM) respectively in normal nasopharyngeal epithelium (NE), tissue adjacent to carcinoma (ATC) and nasopharyngeal carcinoma (NPC). The results showed that all 20 patients with NE show ed diploid, whereas 38 (38%) of 101 patients with NPC, 7 (58%) of 12 patients with recurrent NPC, 6 (30%) of 20 patients with ATC and 2 (33%) of 6 patients with NE positive to EBVCA-IgA test showed nondiploid. A total of 19 morphometric parameters of the cell were measured and calculated. The results showed that with malignant transformation of NE, most of the cellular morphometric parameters gradually increased in numerical values while no marked difference was shown between normal NE and simple hyperplasia or metaplasia in terms of cytomorphometric characteristics. Most of the parameters in dysplasia changed significantly as compared with those normal NE, simple hyperplasia or metaplasia and NPC groups. Cytomorphome

DNA CONTENT AND MORPHOLOGICAL PARAMETERS IN SYNOVIAL SARCOMA AND SYNOVIOMA

By using image analysis technique with 6 normal synovial tissue specimens as controls, nuclear DNA content and morphological parameters in specimens from 51 synovial sarcomas and 15 synovioma were examined quantitatively, The DNA content in synovial sarcoma and synovioma was significantly different (P < 0.01), and that of synovioma and normal synovial tissue was also different (P < 0.05), Morphological parameters including nuclear areas and perimeters differed significantly among the above three groups (P < 0.01). There was no correlation between the DNA content of synovial sarcoma and its histologic type (P > 0.05), but the DNA content greatly differed between its higher and lower differentiated grade in each type (P < 0.01). The DNA content in synovial sarcoma IIA stage, according to Enneking's staging system, was lower than that in IIB, IIIA and IIIB (P < 0.05), The 5-year survival rate of synovial sarcoma with diploid / nearly diploid (D / ND) pattern was higher than that of aneuploid (AN) pattern (P = 0.028), The study suggests that analysis of the DNA content and morphological parameters of tumor cells is helpful in the diagnosis and pathologic grading of synovial sarcoma, It also provides a guide to clinical operation and estimation of the prognosis, Although synovioma is usually a well-differentiated and benign lesion, this investigation also suggested that increased DNA content may be an indication of poor prognosis for this neoplasm.

Effects ofm perioperative cimetidine administration on tumor cell nuclear morphology and DNA content in patients with gastric and colorectal cancer

Objective To explore the effects of perioperative cimetidine administration on tumor cell nuclear morphometric parameters and DNA content in patients with gastric and colorectal adenocarcinoma (GCRA) Past studies have attributed the antitumor effect of cimetidine to its immunomodulatory property, which led to an increase of cellular immunity Whether there are other possible mechanisms by which cimetidine exerts its antitumor function is unknown 49 patients with GCRA were randomized into treatment group (n=25) and control group (n=24) based on whether cimetidine was applied to them during the perioperative periold The treatment group started oral cimetidine intake 400mg, tid, 7-10d before oiperation, followed by curative surgery 'The control group did not receive cimetidine Tumor specimens were paraffin embedded for 4μm thick microsection and stained with (1) hematoxylin and eosin (HE) for the morphometric measurements of tumor cell nuclear area (NA), nuclear perimeter (NP), maximal nuclear diameter (MMND) and minimal nuclea4r diameter (MNND); (2) feulgen stain for tumor nuclear DNA content analysis by IBAS Image Analyzer The percentages (%) of diploidy (2C), tripletetraploidy (3C 4C), quintuple ploidy (5C) and >quintuple ploidy (>5C) tumor cells were calculated, using the mean value of DNA content of 50 lymphocytes as normal 2C control 3C 5C cells were designated as law aneuploid cells and >5C cells as high aneupoid cells Results The clinicopathological variables between the two groups were balanced and comparable There were no statistically significant differences between bthe treatment and control groups in regard of the following parameters: age, gender, tumor location, pathological type, TNM stage, and degree of differentiation The NA (μm 2), NP (μm), MMND (μm) and MNND (μm) for treatm ent group/control group were 23 54±5 08/34 698±10 18 ( P <0 001), 22 06±4 43/24 88±4 05 ( P <0 05),7 84±1 64/8 62±1 24 ( P >0 05), and 4 42±0 61/5 41±0 89 ( P <0 001), respectively The percentages (%) of 2C, 3C 4C, 5C and >5C tumor cells for treatment group/control group were 16 64±2 58/5 35±2 14 ( P <0 002), 39 84±2 28/35 70±3 58 ( P >0 50), 12 42±5 00/14 48±0 74 ( P >0 20), 31 11±6 86/45 97±3 82 ( P <0 005), respectively In the treatment group, there was a tendency tiowards low aneuploid tumor cells from high an euploid tumor cells However, high aneuploid tumor cells predominated in the control group Conclusion Perioperative administration of cimetidine to GCRA patients could decrease the size of tumor cell nuclei, raise the percentage of diploid tumor cells, and partially convery high aneuploid tumor cells into low aneuploid tumor cells All of these effects may in turn help reduce the proliferative potential and invasiveness of tumor cells The direct inhibitory functions on tumor cell nuclei may be a new antitumor mechanism of cimetidine, in addition to its immunomodulatory action