OBJECTIVETo detect the expression levels of programmed death-1 (PD-1) and programmed death ligand-1 (PD-L1) in the peripheral blood of patients with oral squamous cell carcinoma (OSCC) and to discuss their biological and clinical significance.

METHODSPD-1/PD-L1 expression on the surface of T-lymphocytes and the counts of T-lymphocyte subpopulations of peripheral blood in 82 patients with OSCC (OSCC group) and 25 healthy controls (control group) were examined via flow cytometry. The expression levels of soluble PD-1 (sPD-1) and soluble PD-L1 (sPD-L1) in the serum were observed through enzyme-link immunology method. The data were tested and analyzed with SPSS 17.0 software.

RESULTSThe percentage of CD8+ T cells in the OSCC group was significantly higher than that in the control group (P<0.05), whereas the percentages of CD3+ and CD4+ T cells as well as CD4+/CD8+ ratio were significantly lower than those in the control group (P<0.05). The positive rates of PD-1 and PD-L1 in CD3+ and CD4+ T cells in OSCC peripheral blood were remarkably higher than those in the control group (P<0.01). Difference was not observed between the expression levels of sPD-1 in the serum of OSCC group and those in the control group (P>0.05), but the average of sPD-L1 was remarkably higher than that in the control group (P<0.05). sPD-L1 expression was related to clinical stage, tumor cell differentiation, and lymph node status (P<0.05) but not related to sex, age, tumor location, and tumor size.

CONCLUSIONT-lymphocyte subpopulations in the peripheral blood of patients with OSCC developed immunosuppression with different degrees. PD-1 and PD-L1 expression levels on the surface of CD3+ and CD4+ T cells significantly increased. Abnormal increase in sPD-L1 expression may be associated with OSCC development.

B7-H1 Antigen ; metabolism ; Carcinoma, Squamous Cell ; metabolism ; Case-Control Studies ; Flow Cytometry ; Humans ; Mouth Neoplasms ; metabolism ; T-Lymphocyte Subsets

OBJECTIVETo analyze the existence and the dynamic cell frequencies of human cells in goats transplanted in utero with human hematopoietic stem cell (hHSC) by using fluorescence in situ hybridization (FISH) technique.

METHODSInterphase FISH (IFISH) with human-specific 17-chromosome satellite DNA and/or human-specific Y-chromosome satellite DNA as probes was performed to analyze the presence and proportions of human cells in 13 transplanted goats. Samples were peripheral blood cells, bone marrow smears and liver touch imprint preparations.

RESULTSOf the 13 transplanted goats, eleven were identified to present human cells. Among them, two goats transplanted with human male HSC were found to have human male cells. The results demonstrated that these transplanted goats were human/goat HSC xenogeneic chimeras. Human cell frequencies decreased with the goat age (months), but the longest survival reached 21 months. During the detected life periods of goats, human cell frequencies in peripheral blood, bone marrow and liver tissues were less than 1@1000, but local human cell frequencies of 207.92@1000 and 392.41@1000 were detected in the liver tissues of 2 transplanted goats.

CONCLUSIONSThe existence and long-term survival of human cells in transplanted goats detected by FISH indicated that goats were appropriate recipients for hHSC in utero transplantation. The lower human cell frequencies in blood and bone marrow, and the higher local human cell frequencies in liver tissues suggested that the microenvironment of goat liver tissues might favor the survival, proliferation and differentiation of human cells.

Animals ; Female ; Goats ; Hematopoietic Stem Cell Transplantation ; Humans ; In Situ Hybridization, Fluorescence ; Male ; Transplantation, Heterologous ; Uterus ; surgery

AIM: To investigate the mechanism of Hexue Mingmu Tablets(HXMMT)in improving diabetic retinopathy(DR)based on transcriptomics.METHODS: Zebrafish DR models were established by 3-day glucose induction(130 mmol/L)starting at 3 days post-fertilization(dpf). Larvae were randomized into four groups: control group(CG; aquaculture water), model group(MG; 130 mmol/L glucose), low-dose HXMMT treatment group(L-HX; 130 mmol/L glucose +7.5 mg/L HXMMT), and high-dose HXMMT treatment group(H-HX; 130 mmol/L glucose +75 mg/L HXMMT), with a 3-day intervention period until 6 dpf. The area and length of eyes, and body length of zebrafish were observed by stereomicroscopy, retinal morphology was observed by hematoxylin-eosin staining(HE), and retinal vessel diameter was observed under fluorescence microscope. Differentially expressed genes(DEGs)were identified by RNA-sequencing(RNA-seq)technology to further elucidate the molecular mechanism of HXMMT in improving DR in zebrafish, and the sequencing accuracy was validated through quantitative real-time polymerase chain reaction(qRT-PCR).RESULTS: HE staining demonstrated that the intervention with HXMMT significantly improved the disordered cell arrangement, widened gaps, and thickened inner nuclear layer(INL)in ganglion cell layer GCL); retinal vascular diameter quantification revealed that the retinal vessel diameter of the MG significantly increased compared with the CG, and it was significantly changed after the intervention of HXMMT, with significant efficacy in the H-HX(P<0.05); transcriptomics profiling identified 1 470 reversed DEGs, predominantly enriched in the AMPK signaling pathway, FoxO signaling pathway, retinal developmental processes, and tight junction regulation. Technical validation confirmed strong correlation between qRT-PCR and RNA-seq data(R2=0.8571, P<0.05).CONCLUSION: HXMMT may improve retinal vascular microcirculation disorders in DR by regulating core targets including vsx1, pde6c, arr3a, plk1, fbp1b, foxo1a, pcna, and cdk1, as well as synergistically modulating processes such as retinal development in camera-type eyes, visual perception, microtubule cytoskeletal organization, tight junctions, and the AMPK signaling pathway, Foxo signaling pathway.