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Applications of PBK/TOPK in diagnosis and treatment of intimal slim diseases

A thin, diagnostic reagent technology, applied in the direction of disease diagnosis, biochemical equipment and methods, microbial determination/inspection, etc.

Active Publication Date: 2021-10-19
NANJING DRUM TOWER HOSPITAL +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Whether PBK / TOPK is associated with thin intima has not been reported so far

Method used

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  • Applications of PBK/TOPK in diagnosis and treatment of intimal slim diseases
  • Applications of PBK/TOPK in diagnosis and treatment of intimal slim diseases
  • Applications of PBK/TOPK in diagnosis and treatment of intimal slim diseases

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0015] Example 1 Analysis of RNA-seq differentially expressed genes using bioinformatics methods

[0016] 1. Materials, reagents, equipment

[0017] 1.1 Source of human endometrial tissue

[0018] The endometrial tissues of 7 cases of control normal endometrium and 7 cases of patients with thin endometrium were collected. When obtaining endometrial tissues, according to the hormone level, it was ensured that all the stages of endometrial tissue acquisition were unified into the late stage of proliferation. All participants signed informed consent, which was approved by the Ethics Committee of Nanjing Drum Tower Hospital.

[0019] 1.2 Main reagents

[0020] RNasy Plus Micro Kit (Qiagen, Germany), absolute ethanol, DEPC water, enzyme-free water, reverse transcription reagent (Yase), SYBR Green (Yase).

[0021] 1.3 Main instruments

[0022] Qubit 3.0 Fluorometer, Agilent 2100 Bioanalyzer, Illumina Hiseq X10 Platform (Illumina, California, USA), Nano Drop.

[0023] 1.4 Main m...

Embodiment 2

[0030] Example 2 Expression of PBK / TOPK in Endometrial Tissue of Patients with Normal and Thin Endometrium

[0031] 1. Materials, reagents, equipment

[0032] 1.1 Source of human endometrial tissue

[0033] Collect 8 cases of normal endometrial tissue and 10 cases of endometrial tissue from patients with thin endometrium. When obtaining endometrial tissue, according to the hormone level, ensure that the endometrial tissue acquisition stage is unified into the late proliferation stage. All participants signed informed consent, which was approved by the Ethics Committee of Nanjing Drum Tower Hospital.

[0034] 1.2 Main reagents

[0035] TRNzol (Tiangen), chloroform, isopropanol, absolute ethanol, DEPC water, enzyme-free water, reverse transcription reagent (Yase), SYBR Green (Yase), xylene, 95%, 80% and 75% Ethanol, 3% hydrogen peroxide, citric acid repair solution, PBK / TOPK antibody (abcam), DAB chromogenic solution, hematoxylin, ammonia water, neutral resin.

[0036] PBK / T...

Embodiment 3

[0048] Example 3 Effect of PBK / TOPK on proliferation of human endometrial stromal cells

[0049] 1. Materials, reagents, equipment

[0050] 1.1 Main reagents

[0051]CCK8 kit (Vazyme), hyaluronidase (Sigma), DNAase (Roche), PBS, DMEM / F12 medium (Gibco), FBS serum (Gibco), antibiotics, trypsin 1.2 Main instruments

[0052] Filter, centrifuge, cell incubator, cell counter, microplate reader

[0053] 1.3 Main method

[0054] 1.3.1 Isolation and culture of human endometrial stromal cells

[0055] The human endometrial tissue scraped under the hysteroscope was taken, rinsed with normal saline, and cut into pieces. Add 0.1% type Ⅰ collagenase digestion solution, and digest at a constant temperature of 37°C for 1 hour. Pass through a 100-mesh sieve to remove tissue residues, centrifuge the filtrate at 230×g for 5 minutes, and discard the supernatant. Resuspend the pellet in serum-free DMEM / F12, pass through a 400-mesh sieve, centrifuge at 230×g for 5 minutes, and discard the su...

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Abstract

The invention discloses applications of PBK / TOPK in preparation of a reagent for diagnosis and treatment of thin intimal diseases. The applications comprise an application of the PBK / TOPK as a detection target to be applied to preparation of a thin inner membrane diagnostic reagent, and an application of the PBK / TOPK in preparation of drugs for treating diseases caused by thin endometrium. Sequencing and bioinformatics method analysis find that down-regulated genes of patients with thin intima are mainly enriched in cell proliferation, cell cycle and other biological processes. Experiments prove that PBK / TOPK expression in endometrial tissues of a thin endometrial patient is remarkably reduced, and particularly, the reduction of interstitial cells is most remarkable. In-vitro results show that the cell proliferation activity is reduced after PBK / TOPK interference. Therefore, the PBK / TOPK may play an important role in detection and treatment of the thin intima, and can be used for clinical diagnosis and treatment of the thin intima.

Description

technical field [0001] The invention belongs to the field of biopharmaceuticals and relates to the application of PBK / TOPK in the diagnosis and treatment of thin endometrium. Background technique [0002] Embryo implantation is the key to successful pregnancy. Successful implantation requires a well-developed embryo and a well-receptive endometrium. The endometrium is composed of a single layer of columnar epithelial cells and a lamina propria rich in mesenchymal cells. An appropriate thickness of the endometrium is necessary for the implantation of fertilized eggs. Thin endometrium (thin endometrium, TE) means that the patient's endometrium thickness does not increase significantly after being stimulated with high doses of estrogen. However, the definition of thin endometrial thickness has not yet reached a broad consensus. It is generally believed that in the mid-luteal phase (6-10 days after ovulation), the endometrial thickness is <7mm. Its main clinical feature is ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/6883G01N33/573
CPCC12Q1/6883G01N33/573C12Q2600/158G01N2333/912G01N2800/368G01N2800/36
Inventor 胡娅莉赵光锋朱祺姜佩佩戴建武
Owner NANJING DRUM TOWER HOSPITAL