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Ting Lei

Publications and source records attributed to Ting Lei.

12 recordsLinked to original sources

Label-Free Urinary Proteomics Uncovers Immune-Related Non-Invasive Biomarkers for Primary Biliary Cholangitis.

Diagnosis of primary biliary cholangitis (PBC) currently depends on invasive liver biopsy or serum markers with inadequate diagnostic performance. This study aimed to identify non-invasive urinary protein biomarkers for PBC detection. Urine specimens from biopsy-verified PBC patients and healthy controls were processed through ultracentrifugation-based protein extraction, enzymatic digestion, and HPLC-ESI-IT/MS proteomic profiling; protein quantification was completed using Spectronaut v14.8. We identified 194 differentially expressed urinary proteins (109 upregulated, 85 downregulated) and screened 10 immune-related candidates through GO and KEGG enrichment. Pearson correlation further filtered three core proteins, osteopontin (SPP1/OPN), RAMP3 and S100A8, that correlated significantly with key PBC biochemical indices (ALP, GGT, AST, ALT, IgM, p < 0.05). Elevated urinary concentrations of OPN, RAMP3 and S100A8 were validated by ELISA in an independent cohort containing 30 PBC patients and 20 healthy volunteers. In summary, urinary OPN, RAMP3 and S100A8 are markedly increased in PBC patients and hold promise as non-invasive diagnostic biomarkers for PBC; however, their diagnostic specificity against other cholestatic and autoimmune liver diseases remains to be evaluated, and further confirmation in larger multicenter cohorts with disease control groups is warranted.

Humans↗

Cloning, comparative characterization of porcine SCAP gene, and identification of its two splice variants.

Sterol responsive element binding protein (SREBP) cleavage-activating protein (SCAP) is the key regulator of activation of SREBPs, which stimulate most enzymes in cholesterol and lipid synthesis. In order to investigate the molecular basis of lipid metabolism in the pig, a unique model for fat deposition, we isolated and characterized the porcine SCAP. The 4,096-bp full-length porcine SCAP cDNA contains an open reading frame of 3,840 bp. The predicted SCAP protein consists of 1,280 amino acids of 55-92% identity with its vertebrate counterparts. The porcine SCAP gene consists of at least 19 exons and 18 introns, which span over 13 kb of the genome. The porcine SCAP gene was mapped to chromosome 13q21-22 using a porcine-rodent somatic cell hybrid panel. Comparison of SCAP genomic structures from various species revealed intron losses in porcine, Tetraodon and fugu SCAP, and intron gains in cow and chicken SCAP. Moreover, we isolated two novel splicing SCAP variants with 193-bp (variant 2) in-frame deletion from testis and a variant with 291-bp (variant 3) in-frame deletion from liver and muscle, which may affect the function of the porcine SCAP. In conclusion, the intron gains and losses appear to have contributed to the shape of the modern SCAP family. The splice variants detected, first to be reported in any species, may be involved in the particulars of the fat metabolism in the pig. Our data lay foundation for further study of SCAP function in this species.

Alternative Splicing↗

[Preliminary screening and identification of a peptide that binds specifically to gastric cancers cells with high metastasis to peritoneum].

OBJECTIVE: To screen and identify peptides that binds specifically to gastric cancers cells with high metastasis to peritoneum so as to find appropriate vectors for targeting therapy for cancer. METHODS: Human gastric cancer cells of the line GC9811 and those with high metastasis to peritoneum of the line GC9811-P were co-incubated with the 12-mer bacteriophage random peptide library. After 3 round of repeated screening, phage clones were collected. Forty internalized phage single-stranded DNA that specifically binding to the GC98112-P cells were sequenced. GC9811 and GC9811-P cells were co-inoculated with 5 peptides with the N end marked with fluorescein isothiocyanate (FITC) and 1 un-related peptide not binding to GC9811 and GC9811-P cells. Fluorescence microscopy, ELISA, and flow cytometry were used to detect the binding activity. BALB/cnu/nu mice were inoculated intraperitoneally with GC9811 and GC9811-P cells and then randomly divided into. 2 equal groups: experimental group, inoculated with the peptide PIII-FITC and control group (inoculated with un-related peptide PC-FITC. Forty-eight hours later the mice were killed and the peritoneum and tumor masses in different organs were collected and under fluorescence microscopy. RESULTS: DNA sequencing showed that 45% (18/45) of the isolated phages displayed repeated sequence SMSIASPYIALE, and SMSI was defined as a conservative motif. Obvious fluorescence was seen in the GC9811-P cells co-incubated with PIII-FITC and weak fluorescence was seen in the GC9811 cells co-incubated with PIII-FITC. Un-marked un-related peptide PC and PIII-FITC did not influenced the fluorescence staining of the GC9811-P cells, however, no fluorescence could be seen in the GC9811-P cells co-incubated with un-marked PIII and PIII-FITC. The fluorescence positive cell rate was 5.9% in the GC9811 cells co-incubated with PIII-FITC, and was 90.2% in the GC9811-P cells co-incubated with PIII-FITC. The fluorescence positive cell rates of the GC9811 cells and GC9811-P cells co-incubated with PC-FITC were 10.1% and 9.9% respectively 10.1% and 9.9% respectively. The fluorescence strength of the GC9811-P cells co-incubated with PIII-FITC was significantly greater than that of the GC9811-P cells co-incubated with PC-FTIC at any time-point and dose (all P < 0.01), and increased along with the increase of co-incubation time and dose of PIII-FITC. The peritoneal tumor tissues caused by the GC9811-P cells of the mice showed strong fluorescence and those caused by GC9811 cells only showed very weak fluorescence. Weak fluorescence could be seen in the tumor masses in the lymph nodes, liver, and muscle of the mice inoculated with GC9811-P cells and was not seen in the tissues of the mice inoculated with GC9811 cells. CONCLUSION: The sequence SMSIASPYIALE that specifically binds to human gastric cancer cells with high metastasis has been screened that has the potential to be used as a marker and targeting vector in diagnosis and treatment of gastric cancer.

Amino Acid Sequence↗

Expression of EGFR and LRIG-1 in human trigeminal neurinoma.

The expression of epidermal growth factor receptor (EGFR) and leucine-rich repeats and immunoglobulin-like domain 1 (LRIG-1) in human trigeminal neurinoma was investigated and their effect on the origination and development of trigeminal neurinoma, and the relationship between them was studied. By using immunohistochemistry with tissue chip, the expression of EGFR and LRIG-1 was detected in 23 cases of trigeminal neurinoma. It was found that in the 23 cases, the expression rate of EGFR was 21.74%, while that of the LRIG-1 was 78.26%. There was a negative correlation between them. It was suggested that LRIG-1 might inhibit the malignant differentiation and proliferation of the trigeminal neurinoma possibly by the negative feedback loop of EGFR.

Adult↗

Treatment and prognosis of pituitary adenomas in children.

In order to explore the treatments and prognosis of pituitary adenomas in children, the experience in the treatment of 15 children with pituitary adenomas was summed up. The clinical data of 15 children with pituitary adenomas in Tongji Hospital were retrospectively analyzed from Jan. 1997 to Aug. 2004, including 9 males and 6 females with the age ranging from 9 - 15 years old (mean 12.8). The disease duration was 3 months to 5 years (mean 2 years). Fourteen cases were subjected to the microsurgical operation through oral tanssphenoidal approach and one case to medical treatment. All cases were followed up from 1 to 5 years by imaging studies and endocrine investigation. The results showed that 13 cases obtained total removal of the tumor and one subtotal removal of tumor. No severe complications and operation death occurred. It was concluded that the tanssphenoidal approach was the optimal treatment for children with pituitary adenomas. Regular and long-term follow-up is of great importance.

Adenoma↗

Effect and mechanism of epidermal growth factor on proliferation of GL15 gliomas cell line.

The effects of epidermal growth factor (EGF) on proliferation of G15 glioma cells and the possible mechanisms were investigated. GFAP and EGFR expression was detected by immunohistochemical method. After the cells were treated with EGF at different concentrations, cell count method was used to determine the proliferation of glioma cells, cell cycle and apoptosis were analyzed by flow cytometry (FCM), and laser scan confocal microscope (LSCM) was used to measure the cytoplasmic free calcium. The results showed that GFAP was diffusedly expressed in GL15 cells and EGFR was over-expressed. EGF at doses of < or =1 ng/mL could significantly stimulate cell proliferation, cells in phase G0/G1 decreased, and those in phase S increased. EGF at doses of 10 and 100 ng/ml could inhibit the cell proliferation significantly, and the apoptosis ratio in high dose of EGF group was higher than in control group. EGF could significantly induce a quick rise of intracellular free calcium, but the peak value of intracellular free calcium activated by high dose of EGF was higher than by low dose of EGF. It was suggested that EGF had a dual effect on gliomas: low dose of EGF could stimulate the cell proliferation of gliomas, but high dose of EGF could induce the cell apoptosis and inhibit the proliferation of gliomas, which might be contributed to the difference of intracellular free calcium.

Apoptosis↗

Conus medullaris schistosomiasis.

OBJECT: The authors performed a study to investigate the clinical manifestations, treatment strategies, and possible pathogenesis of conus medullaris schistosomiasis. METHODS: Six cases collected from the authors' experience and four cases reported in the literature were studied retrospectively for clinical manifestations, treatment outcomes, and prognosis. All patients experienced progressive lower-extremity weakness and functional bowel and bladder impairment. Although the magnetic resonance (MR) imaging results suggested the presence of a conus medullaris tumor, schistosomiasis was diagnosed based on pathological results obtained in the 10 patients. The results of surgery followed by pyquiton and hormone treatment confirmed the diagnosis, and the patients' prognoses were good. CONCLUSIONS: This pathological entity is predominantly found in adults, and the clinical manifestations have no specificity, although the MR imaging may provide some clues. As a form of ectopic schistosomiasis, conus medullaris schistosomiasis deserves special consideration and further exploration. If an early diagnosis can be made and pyquiton and hormone therapy is given, surgery can be avoided and the prognosis will remain good.

Adult↗

Expression of human papillomavirus type 16 L1 protein in transgenic tobacco plants.

To develop a plant expression system for the production of the human papillomavirus type 16 (HPV16) vaccine, we investigated whether the HPV16 L1 protein can be expressed in tobacco plants and whether it can be used as the cheapest form of edible vaccine. The HPV16 L1 coding sequence was amplified by PCR using specific primers from the plasmid pGEM-T-HPV16 containing the template sequence, and subcloned into the intermediate vector pUCmT and binary vector pBI121 consecutively to obtain the plant expression plasmid pBI-L1. The T-DNA regions of the pBI-L1 binary vector contained the constitutive Cauliflower mosaic virus (CaMV) 35S promoter and the neomycin phosphotransferase npt II gene, which allowed the selection of transformed plants using kanamycin. The tobacco plants were transformed by co-cultivating them, using the leaf disc method, with Agrobacterium tumefaciens LBA4404, which harbored the plant expression plasmid. The regenerated transgenic tobacco plants were selected using kanamycin, and confirmed by PCR. The results of the Southern blot assay also showed that the HPV16 L1 gene was integrated stably into the genome of the transformed tobacco plants. The Western blot analysis showed that the transformed tobacco leaves could express the HPV16 L1 protein. Furthermore, it was demonstrated by ELISA assay that the expressed protein accounted for 0.034%-0.076% of the total soluble leaf protein, was able to form 55 nm virus-like particles compatible with HPV virus-like particle (VLP), and induced mouse erythrocyte hemagglutination in vitro. The present results indicate that the HPV16 L1 protein can be expressed in transgenic tobacco plants and the expressed protein possesses the natural features of the HPV16 L1 protein, implying that the HPV16 L1 transgenic plants can be potentially used as an edible vaccine.

Agrobacterium tumefaciens↗

[Molecular mechanism of LRIG1 cDNA-induced apoptosis in human glioma cell line H4].

BACKGROUND & OBJECTIVE: Leucine-rich repeats and immunoglobulin-like domains 1(LRIG1)is a kind of transmembrane glycoprotein,which is induced by epidermal growth factor (EGF),and develops inhibitory negative feedback by specific binding with epidermal growth factor receptor (EGFR). This research was to explore the molecular mechanism of LRIG1 inhibiting EGFR signal pathway. METHODS: Plasmid p3XFLAG-CMV9-LRIG1 was transfected into neuroglioma cell line H4. Changes of LRIG1 and EGFR expression at mRNA and protein levels were measured by RT-PCR and Western blot, respectively. Changes of cell proliferation and apoptosis were analyzed by MTT and flow cytometry methods. RESULTS: LRIG1 mRNA level in p3XFLAG-CMV9-LRIG1 transfected H4 cells (1.997+/-0.114) was significantly higher than that in control H4 cells (0.500+/-0.031),and p3XFLAG-CMV9 transfected group (0.357+/-0.035) (P< 0.001). LRIG1 protein level in p3XFLAG-CMV9-LRIG1 transfected H4 cells (1.790+/-0.119) was significantly higher than that in control H4 cells (0.717+/-0.038, P< 0.001), and p3XFLAG-CMV9 transfected H4 cells (0.930+/-0.076, P=0.001). EGFR mRNA level in p3XFLAG-CMV9-LRIG1 transfected H4 cells (0.463+/-0.033) was significantly lower than that in control H4 cells (1.157+/-0.067, P< 0.001), and p3XFLAG-CMV9 transfected H4 cells (0.933+/-0.058, P=0.002). EGFR protein level in p3XFLAG-CMV9-LRIG1 transfected H4 cells (0.703+/-0.067) was significantly lower than that in control H4 cells (1.280+/-0.078, P=0.003),and p3XFLAG-CMV9 transfected H4 cells (1.163+/-0.068,P=0.009). Apoptosis rate in LRIG1-transfected H4 cells (18.89%)was lower than that in control H4 cells (3.11%), and vector-transfected H4 cells (5.42%, P< 0.001). CONCLUSION: LRIG1 participates in construction of negative feedback loop of EGFR, which may inhibits growth of glioma cells.

Apoptosis↗

Anti-endotoxic effects of syringic acid of Radix Isatidis.

The anti-endotoxic effect of syringic acid (SA) isolated from Radix Isatidis (Banlangen, BLG) was studied. SA was extracted and isolated from BLG and diluted into 1% solution. The content of SA-pretreated endotoxin (ET) was quantitatively determined using Limulus test. The ability of fever induction of ET pretreated with SA was measured using endotoxin-induced fever test in rabbits. The LPS-induced death in mice pretreated with and without SA was compared. Results showed that after pretreatment with SA, 83.16% of ET was destroyed, the ET-induced fever in rabbits relieved markedly and the LPS-induced death rate in mice dropped from 68% to 20%. It was concluded that SA isolated from BLG had anti-endotoxic effects.

Animals↗

[Basic study of biodegradable controlled release chemotherapy].

This study sought to assess the biocompatibility of P(DA-SA)-Adriamycin, a new controlled-release chemotherapy system, in rabbit brain, and to examine its controlled release effect both in vitro and in vivo and its curative effects in vitro. The reaction of animal brain to the implanted P(DA-SA) or P(DA-SA)-Adriamycin was observed. The controlled-release profiles in phosphate buffer solutions and in rabbit brain were measured by UV spectrometry. Then, through flow cytometer, the rate of apoptosis in cultured glioma cells was tested. The reaction of rabbit brain to P(DA-SA) polymer was moderate and not significantly different from that to Gelfoam. The controlled-release rate of P(DA-SA)-Adriamycin in vitro and in vivo was stable and the duration of controlled-release of P(DA-SA)-Adriamycin spanned three weeks. The rate for apoptosis of glioma cells of P(DA-SA)-Adriamycin group was 69.9%, which was significantly higher than that of the control group. In conclusion, P (DA-SA)-Adriamycin controlled release chemotherapy system that bears curative effect has favorable controlled-release effect and good biocompatibility in rabbit brain. This system has potential value in treatment of malignant brain tumor.

Animals↗