Human il 10 elisa kit

Post treatment, the cultural supernatants of HUVECs in each group were collected and centrifuged at 1000 rpm for 2 min to remove cell debris. Supernatants were harvested for cytokine detection of MCP-1, sICAM-1, and sVCAM-1. The operation steps were strictly carried out according to the instructions of the ELISA kit (human MCP-1 ELISA kit: ab179886, Abcam; human sICAM-1 ELISA kit: 70-EK189-96, MultiSciences; human sVCAM-1 ELISA kit: DVC00, R&D Systems). In addition, ELISA was applied to detect M2 macrophage marker molecules (IL-10) and M1 macrophage marker molecules (IL-6 and TNF-α) in their corresponding supernatants, respectively, as per the instructions of the ELISA kit (human IL-10 ELISA kit: SEKH-0018; Solarbio; human IL-6 ELISA kit: SEKH-001; Solarbio; human TNF-α ELISA kit: SEKH-0047; Solarbio).

Collected vaginal secretion was smeared, naturally dried, fixed, and stained with Gram's crystal violet solution. After staining, bacterial diversity and Lactobacillus density were observed under oil immersion field microscope. Bacterial density was divided into four levels according to the number of bacteria: grade 1 (+) as 1–3 kinds of bacterium; grade 2 (++) as 4–6 kinds of bacterium; grade 3 (+++) as 7–10 kinds of bacterium; and grade 4 (++++) as more than 10 kinds of bacterium. Lactobacillus concentration was categorized into four levels according to the number of Lactobacilli in each field of view, and 5–8 fields were observed. No Lactobacillus was recorded as (-); less than 1 was recorded as (+); 1–5 was recorded as (++); 6–30 was recorded as (+++); and more than 30 was recorded as (++++). Immune factors (IL-2, IL-10) level in vaginal lavage fluid were analyzed using Human IL-2 ELISA KIT (SEKH-0008, Solarbio, Beijing, China) and Human IL-10 ELISA KIT(SEKH-0018, Solarbio, Beijing, China) by microplate reader (DNM-9602, Beijing, China).
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