Limiting dilution assay and ~(51)Cr-release assay were used to quantitate the pre-cursors of MAF-secreting T cells in lymph nodes or lymphoblast population of themice infected with Leishmania major(Lm).At lst and 4th week after infectionwith live Lm,the numbers of MAF-P in lymph nodes of resistant mice(CBA/T6)and susceptible mice(BALB/C)showed no significant difference but resistantmice had much more MAF-P than susceptible mice at 8th week.The effect ofantigen,interleukin-2 and irradiated syngeneic spleen cells on the detection of MAF-P were investigated、It was found that syngeneic spleen cells were an impor-tant factors for the assay of MAF-P of lymphoblasts but lymph nodes cells,and that the addition of both antigen and interleukin-2 can significantly increasethe number of MAF-P than antigen or interleukin-2 alone when the cells oflymph nodes were examined.

Renal cell carcinoma is one of the significant diseases endangering human health. Recent findings have shown that the human microbiome plays an important role in the occurrence and development of renal cell carcinoma, influencing its regression and treatment outcome. At present, microecological research on renal cell carcinoma are still in their initial stages, and their regulatory roles and specific mechanisms still need to be further explored. This article reviews the relationship between the human microbiome and renal cell carcinoma occurrence and development, as well as its role in diagnosis and therapies.

ObjectiveTo investigate the potential mechanism of Shenqi Yiliu Formula （参芪抑瘤方） in treating precancerous lesions of gastric cancer （PLGC） by the Wnt/β-catenin signaling pathway. MethodsThe CCK-8 assay was used to determine the optimal intervention time for Shenqi Yiliu Formula drug-containing serum and the concentration of the Wnt/β-catenin pathway inhibitor XAV939 depends on the survival rate of AGS gastric cancer cell line. AGS cells were divided into the gastric cancer cell group （15% blank serum）， inhibitor group （selected concentration of XAV939）， high-dose Shenqi Yiliu Formula group （12% Shenqi Yiliu Formula drug-containing serum + 3% blank serum）， medium-dose Shenqi Yiliu Formula group （6% Shenqi Yiliu Formula drug-containing serum + 9% blank serum）， and low-dose Shenqi Yiliu Formula group （3% Shenqi Yiliu Formula drug-containing serum + 12% blank serum）. Each group was tested in triplicate. After culturing for 24 and 48 hours， cell migration and invasion were assessed by scratch assays； after a selected intervention period （48 hours）， cell cycle distribution was analyzed using flow cytometry， Ki67 protein levels were detected by immunofluorescence， the protein levels of Wnt， β-catenin， GSK-3β， and intranuclear T-cell specific factor（TCF） were measured by the protein immunoblotting assay， and the mRNA expressions of these above factors were determined by quantitative real-time PCR. ResultsThe optimal intervention time for Shenqi Yiliu Formula drug-containing serum was determined to be 48 hours， and the effective concentration of XAV939 was 20 μmol/L. Compared with the gastric cancer cell group， Shenqi Yiliu Formula at all doses reduced the cell migration rate at 24 and 48 hours （P＜0.05）， except for the low-dose group at 24 hours. Compared to the low-dose group at corresponding time points， high- and medium-dose Shenqi Yiliu Formula groups showed significantly reduced migration rates， particularly the high-dose group at 48 hours （P＜0.05）. Compared with the gastric cancer cell group， the high-dose Shenqi Yiliu Formula and inhibitor groups exhibited reduced protein and mRNA levels of Wnt， β-catenin， and TCF， along with reduced Ki67 protein levels and a decreased proportion of cells in the S and G2 phases of the cell cycle， but GSK-3β protein levels， GSK-3β mRNA expression， and the proportion of cells in the G1 phase increased （P＜0.05）. Compared to the inhibitor group， the high-dose Shenqi Yiliu Formula group showed a decreased proportion of G1-phase cells and an increased proportion of G2-phase cells （P＜0.05）， although differences in Wnt and β-catenin protein levels and mRNA expressions were not statistically significant （P＞0.05）. ConclusionShenqi Yiliu Formula drug-containing serum inhibits the migration and invasion of gastric cancer AGS cells and block the cell cycle at G1 phase， and its underlying mechanism may be related to the regulation of the Wnt/β-catenin signaling pathway.