1.Alpha-galactosylceramide loaded tumor cells in combination with TLR9 agonists induce potent anti-tumor responses in a murine model of colon cancer
Tiangeng DONG ; Tuo YI ; Xinqiang HONG ; Mengxuan YANG ; Shengli LIN ; Xingyuan XU ; Wenxiang LI ; Weixin NIU
Chinese Journal of Microbiology and Immunology 2014;(7):491-497
Objective To design a new cancer vaccine by using alpha-galactosylceramide (α-Galcer,α-GC) loaded tumor cells in combination with TLR 9 ligand and to evaluate its therapeutic effects on colon canc-er in mice.Methods MC38 cells were transfected with lentivirus (GFP-CD1d) to prepare CD1d-MC38 cells. The expression of CD1d molecules in CD1d-MC38 cells was detected by fluorescence microscopy , RT-PCR and flow cytometry.The sorted CD1d-MC38 cells were loaded with α-Galcer to prepare CD1d-MC38/α-GC complex. Flow cytometry was performed to evaluate the efficiency of combination .A mouse model of colon cancer was es-tablished to investigate the therapeutic effects of α-Galcer loaded tumor cells in combination with TLR 9 ligand ( CD1d-MC38/α-GC+CpG1826) on colon cancer in mice by analyzing tumor growth and mice survival time .Im-munohistochemical staining was used to detect CD 4+T and CD8+T infiltrating lymphocytes in tumor tissues .Re-sults The MC38 cancer cells that expressed CD 1d and GFP were successfully constructed , among which 98.10%±2.53%were positive for CD1d.Moreover, the CD1d-MC38 cells could combine with α-Galcer effec-tively in a dose and time dependent manner .Compared with PBS treated group ,α-GC treated group and TLR9 ligand treated group , the experimental vaccine strategy was sufficient to inhibit the growth of established tumors and prolong survival of tumor-bearing mice (P<0.01).Immunohistochemistry analysis revealed that levels of CD4+T cells and CD8+T cells in experiment group were significantly higher than those in groups treated with PBS,α-GC and TLR9 ligand (P<0.01).Conclusion CD1d-MC38/α-GC in combination with CpG1826 could efficiently inhibit the growth of established tumors and prolong survival of tumor-bearing mice .Immunohisto-chemistry analysis revealed that CD 4+T cells and CD8+T cells played important roles in anti-tumor immunity.
2.Expression and clinical significance of PI3K and MMP-7 proteins in esophageal squamous cell carcinoma.
Lei WANG ; Xinqiang NIU ; Baoen SHAN ; Ming HE ; Xianli MENG ; Bing ZHANG ; Shijie WANG
Clinical Medicine of China 2009;25(10):1083-1085
Objective To study the expression of PI3K and MMP-7 in esophageal carcinoma and the rela-tionship between the expression of PI3K and MMP-7 and carcinogenesis and progression of esophageal carcinoma. Methods PI3K and MMP-7 expression were detected in 24 normal esophageal mucosa,94 primary tumor tissues with SP immunohistochemistal method. Results There were significant differences of PI3K and MMP-7 expressions between esophageal carcinoma and normal mucesa epithelium ( all P < 0.01 ) [71.28% (67/94) vs 4.17% ( 1/24 ) and 52.13% (49/94) vs 0% (0/24)]. There were significant correlations between PI3K expression and the degrees of differentiation,invasive depth,clinical staging and the metastasis of lymph node (all P <0.01 ). The positive ex-pression rate of MMP-7 had the relationship with metastasis of lymph node (P < 0.05 ), the degrees of differentiation ( P < 0.05 ) invasive depth ( P < 0.01 ), and clinical staging( P < 0.05 ) . There was a positive relationship between PI3K and MMP-7 expression in this study (r = 0. 232 ,P = 0.025). Conlusions PBK and MMP-7 play an impor-tant roles in carcinogenesis and progression of esophageal carcinoma and can be used as valuable biomarkers to eval-uate biological characteristics in esophageal squamous cell carcinoma.
3.Usher syndrome with methylmalonic acidemia and homocysteine in an infant
Yujuan WANG ; Jinjun TIAN ; Linyuan NIU ; Liqin LIU ; Xinqiang LAN
Chinese Journal of Perinatal Medicine 2022;25(12):975-977
A case of Usher syndrome with methylmalonic acidemia and homocysteine is reported. The patient was a two-month-old and small for gestational age male infant hospitalized for "feeding difficulties" during the neonatal period. The baby boy presented hypotonia, microcephaly, and hearing loss after birth. Genetic test found compound heterozygous mutations of c.482G>A and c.567dup in MMACHC, and both were pathogenic mutations inherited from his parents. Moreover, the patient also had compound heterozygous variants at c.2802T>G and c.14017T>C of USH2A gene. The former was suspected to be pathogenic, and the latter was of unknown clinical significance. Both were from the parents. Usher syndrome and methylmalonic acidemia with homocysteine were clinically diagnosed. Followed up to the age of two, the child was found with moderate mental retardation, while the physical development was comparable to that of the same age group.
4.Analysis of GJB2, SLC26A4, GJB3 and 12S rRNA gene mutations among patients with nonsyndromic hearing loss from eastern Shandong.
Shiyu SUN ; Linyuan NIU ; Jinjun TIAN ; Wei CHEN ; Yanna LI ; Ningning XIA ; Caining JYU ; Xiaoli CHEN ; Chunxiao ZHANG ; Xinqiang LAN
Chinese Journal of Medical Genetics 2019;36(5):433-438
OBJECTIVE:
To explore the characteristics of mutations of four common pathogenic genes (GJB2, SLC26A4, GJB3 and 12S rRNA) among patients with nonsyndromic hearing loss (NSHL) from eastern Shandong.
METHODS:
Peripheral blood samples of 420 NSHL patients were collected, and a hereditary-deafness-gene microarray was used to detect GJB2 c.235delC, c.299-300delAT, c.35delG and c.176del16 mutations, GJB3 c.538C>T mutation, SLC26A4 c.2168A>G and c.IVS7-2A>G mutations, and 12S rRNA c.1555A>C and c.1494C>T mutations. For patients carrying single heterozygous mutations, the coding regions of the above genes were analyzed with Sanger sequencing.
RESULTS:
The results of the microarray assay and Sanger sequencing showed that 84 patients (20.00%) carried GJB2 mutations, with c.235delC (16.43%) and c.299-300delAT (7.86%) being most common. Seventy-five patients (17.86%) carried SLC26A4 mutations, for which c.IVS7-2A>G accounted for 15.71%. In addition, 5.95% of patients carried 12S rRNA mutations. Only one patient was found to carried GJB3 mutation (c.538C>T).
CONCLUSION
Common pathogenic mutations for NSHL in eastern Shandong included GJB2 c.235delC and SLC26A4 c.IVS7-2A>G. Of note, 5.95% of patients were due to 12S rRNA m.1555A>G mutation, which gave a frequency greater than other regions of China.
China
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Connexin 26
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Connexins
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DNA Mutational Analysis
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DNA, Mitochondrial
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Deafness
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Genes, rRNA
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Hearing Loss
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Humans
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Mutation
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RNA, Ribosomal
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Sulfate Transporters