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Biomedical subjects

Peng Shang

Publications and source records attributed to Peng Shang.

15 recordsLinked to original sources

CRISPR activation reveals SOX5/6/9 as key transcriptional regulators directing iPSC-derived cells toward a notochordal lineage.

Intervertebral disc (IVD) degeneration, a leading cause of chronic lower back pain, is associated with loss of vacuolated notochordal cells (NCs) and fibrotic remodeling of the nucleus pulposus. Emerging therapies increasingly focus on NCs, which are rare but therapeutically relevant cells for regenerating degenerated IVDs. In this study, we used CRISPR-based transactivation (CRISPRa) to direct the differentiation of human induced pluripotent stem cells (iPSCs) into the NC lineage. We tested CRISPRa-mediated activation of NOTO, TBXT, FOXA2, SOX5, SOX6, and SOX9, coupled with single-cell sequencing of Aggrecan-2A-mScarlet reporter iPSCs. This approach identified the SOX5/6/9 combination (SOX-trio) as critical for promoting NC lineage commitment. The SOX-trio yielded the largest cell population expressing a range of genes previously associated with NC identity, including SHH, FOXA1, FOXA2, FOXJ1, FN1, ALCAM, KRT8, and KRT18. Our study demonstrates the integration of CRISPRa with single-cell technologies as a powerful platform for investigating and enriching iPSC-derived NCs, supporting future regenerative strategies across various fields.

Humans↗

siRNA targeted against HAb18G/CD147 inhibits MMP-2 secretion, actin and FAK expression in hepatocellular carcinoma cell line via ERK1/2 pathway.

HAb18G/CD147 has been identified as a factor that induces MMPs production. SiRNA targeted against HAb18G/CD147 was transfected into FHCC-98 cells (a HCC cell line) to knockdown its expression. The results showed that downregulating HAb18G/CD147 decreased ERK1/2, MMP-2 and FAK levels and inhibited cell motility and invasion, together with rearranged actin stress fiber formation, while had no effects on integrin alpha3beta1 expression. MEK1/2 inhibitor, U0126, inhibited MMP-2, FAK and actin expression in FHCC-98 cell line. The findings indicate that si-HAb18G inhibits gelatinase production, actin and FAK expression in FHCC-98 via an ERK1/2 signaling pathway.

Actins↗

Targeting radioimmunotherapy of hepatocellular carcinoma with iodine (131I) metuximab injection: clinical phase I/II trials.

PURPOSE: HAb18G/CD147 is a hepatocellular carcinoma (HCC)-associated antigen. We developed iodine (131I) metuximab injection (Licartin), a novel 131I-labeled HAb18G/CD147-specific monoclonal antibody Fab'2 fragment, and evaluated its safety, pharmacokinetics, and clinical efficacy on HCC in Phase I/II trials. METHODS AND MATERIALS: In a Phase I trial, 28 patients were randomly assigned to receive the injection in 9.25-, 18.5-, 27.75-, or 37-MBq/kg doses by hepatic artery infusion. In a multicenter Phase II trial, 106 patients received the injection (27.75 MBq/kg) on Day 1 of a 28-day cycle. Response rate and survival rate were the endpoints. RESULTS: No life-threatening toxic effects were found. The safe dosage was 27.75 MBq/kg. The blood clearance fitted a biphasic model, and its half-life was 90.56-63.93 h. In the Phase II trial, the injection was found to be targeted and concentrated to tumor tissues. Of the 73 patients completing two cycles, 6 (8.22%) had a partial response, 14 (19.18%) minor response, and 43 (58.90%) stable disease. The 21-month survival rate was 44.54%. The survival rate of progression-free patients was significantly higher than that of patients with progressive disease after either one or two cycles (p < 0.0001 or p = 0.0019). CONCLUSION: Iodine (131I) metuximab injection is safe and active for HCC patients.

Adolescent↗

Function of HAb18G/CD147 in invasion of host cells by severe acute respiratory syndrome coronavirus.

To identify the function of HAb18G/CD147 in invasion of host cells by severe acute respiratory syndrome (SARS) coronavirus (CoV), we analyzed the protein-protein interaction among HAb18G/CD147, cyclophilin A (CyPA), and SARS-CoV structural proteins by coimmunoprecipitation and surface plasmon resonance analysis. Although none of the SARS-CoV proteins was found to be directly bound to HAb18G/CD147, the nucleocapsid (N) protein of SARS-CoV was bound to CyPA, which interacted with HAb18G/CD147. Further research showed that HAb18G/CD147, a transmembrane molecule, was highly expressed on 293 cells and that CyPA was integrated with SARS-CoV. HAb18G/CD147-antagonistic peptide (AP)-9, an AP of HAb18G/CD147, had a high rate of binding to 293 cells and an inhibitory effect on SARS-CoV. These results show that HAb18G/CD147, mediated by CyPA bound to SARS-CoV N protein, plays a functional role in facilitating invasion of host cells by SARS-CoV. Our findings provide some evidence for the cytologic mechanism of invasion by SARS-CoV and provide a molecular basis for screening anti-SARS drugs.

Antigens, CD↗

[In vitro induction of directional differentiation of bone marrow mesenchymal stem cells towards chondrocytes].

AIM: To induce bone marrow mesenchymal stem cells or bone marrow stroma cells (MSCs) to differentiate directionally towards chondrocytes in vitro and then identify the differentiated cells. METHODS: MSCs were isolated from bone marrows of healthy adult human donors and then the cultured. MSCs at the 3rd passage were induced by TGF-beta1, dexamethasone(Dex) and Vitamin C(Vit C). 14 days later, micro-cell aggregates formed by the induced MSCs were embedded in paraffin, sectioned and stained with HE, toluidine blue or anti-type II collagen mAb (Col II). The expression of Col II in the induced MSCs also were detected by Western blot and RT-PCR. RESULTS: The induced cells exhibited a chondrocyte-like morphology.The staining of toluidine blue and Col II on extracellular matrix in the micro-cell aggregates was positive. Western blot and RT-PCR showed that Col II was only expressed in the induced cells. CONCLUSION: MSCs can differentiate into chondrocytes under the induction of TGF-beta1, Dex and Vit C.

Adult↗

[Effects of N-methyl-D-aspartate receptor in visceral, hypersensitivity in rats with colonic inflammation].

OBJECTIVE: To investigate the effects of N-methyl-D-aspartate receptor (NMDAR) in the spinal dorsal horn in visceral hypersensitivity in rats with colonic inflammation. METHODS: Seventy adult male Sprague-Dawley (SD) rats were randomly divided into the experimental group and the control group. Colonic inflammation was induced in the experimental rats by intraluminal administration of trinitrobenzenesulfonic acid (TNBS). Saline was administered intraluminally in the control rats. After 3, 7, 14, and 28 days of administration, abdominal contractions induced by inflation of a balloon colonically inserted were recorded in rats by implanting electrodes in the abdominal striated muscles. Immunohistochemistry method was used to study the expression of NMDAR1 and NMDAR2A/B in lumbarsacral spinal cord after inflammation. RESULTS: Colonic distension evoked a significant increase of abdominal contractions after 3, 7 and 14 days of TNBS administration. After 28 days of TNBS administration, abdominal contractions were still significantly increased in 2 TNBS-treated rats compared with the control rats. After 7 and 14 days of TNBS administration, NMDAR1 and NMDAR2A/B-immunoreactive cells were significantly increased compared with the control group (P <0.05). Twenty-eight days after TNBS administration, the number of NMDAR1-IR and NMDAR2A/B-IR neurons was still significantly increased in 4 TNBS-treated rats compared with the saline-treated rats (P < 0.05). CONCLUSION: NMDAR was involved in the transmission of visceral nociceptive stimuli. After the remission of colonic inflammation, increased expression of NMDAR1 and NMDAR2A/B in the spinal dorsal horn may induce persistent neuronal hyperactivity, which results in visceral hypersensitivity.

Animals↗

Establishment and characterization of human hepatocellular carcinoma cell line FHCC-98.

AIM: To establish a novel human hepatocellular carcinoma (HCC) cell line FHCC-98 from HCC tissue and to provide a suitable model for studying HCC occurrence, progress and metastasis. METHODS: Serially passaged cells were cultured and their morphologies were observed under light and electron microscope. Cytogenetic study was conducted by using flow cytometry and chromosome analysis. Expressions of tumor markers such as alpha-fetoprotein (AFP), cytokeratin (CK) and hepatoma metastasis-associated factor HAb18G/CD147 on the FHCC-98 cells were detected by immunocytochemistry or Western blotting. Lactic dehydrogenase (LDH) isoenzymes were detected by polyacrylamide gel electrophoresis (PAGE). Xenograft was performed by inoculating FHCC-98 cells into the flanks of nude mice. RESULTS: Morphology of FHCC-98 cells was the same as that of other malignant cells. The expressions of the cells were positive for HAb18G/CD147 and CK, and negative for AFP. Its population doubling time was 21.4 h. The cell DNA was tetraploid and the major chromosomes were triploid by cytogenetics analysis. The tumorigenicity in nude mice was 100%. PAGE showed four bands representing LDH2, LDH3, LDH4 and LDH5. CONCLUSION: FHCC-98 is a novel HCC cell line and an ideal cell model for further exploring the mechanism of hepatocellular carcinoma invasion and metastasis.

Adult↗

[Purification of colorectal cancer-associated antigen by immuno-affinity chromatography].

AIM: To purify and characterize the colorectal cancer-associated antigen from cultured colorectal cancer cells. METHODS: The colorectal cancer cell lines that highly expressed the associated antigen were selected by flow cytometry with five specific monoclonal antibodies (mAbs) CYL1-5. Western blot was used to determine the binding ability of five mAbs to the associated antigens released from colorectal cells lysed with single or triplex-detergent, respectively. The mAb with highest binding ability was employed as the ligand for the immuno-affinity chromatography. The antigens purified through immuno-affinity chromatography were identified by Western blot. RESULTS: The associated antigens were highly expressed on the colorectal cancer cell lines Hce-8693. The binding ability of mAb CYL-2 to the antigen was higher than that of CYL-1, CYL-3-5. The purified associated antigen binding to mAb CYL-2 was a heterodimer composed of two subunits with relative molecular mass (M(r)) of 60 x 10(3) and 70 x 10(3), respectively. CONCLUSION: The purified associated-antigen binding to mAb CYL-2 was obtained from the colorectal cancer cell line Hce-8693 through immuno-affinity chromatography with mAb CYL-2.

Antibodies, Monoclonal↗

[Digital model of human lower extremity musculature based on CT & MRI].

A new method to reconstruct comparatively complete muscle model of human lower limb from CT and MRI data is presented. Topological structure of more than fourteen muscles is built and the coordinates of origin and insertion points are given. Based on this model, straight-line model and centroid-line muscle model are acquired. Muscle force prediction is discussed according to the model reconstructed, and a multi-objective optimization method is put forward for evaluating muscle forces of human lower extremity.

Humans↗

Inhibitory effects of antisense RNA of HAb18G/CD147 on invasion of hepatocellular carcinoma cells in vitro.

AIM: To study the inhibitory effects of antisense RNA of HAb18G/CD147 on invasion of hepatocellular carcinoma(HCC) cells in vitro. METHODS: Antisense RNA of HAb18G/CD147 vector PCI-asHAb18G was constructed by reversely inserting HAb18G/CD147 cDNA to eukaryotic expression vector PCI-neo. The HCC cell line HHCC was transfected by PCI-asHAb18G via cation liposome. Expression of HAb18G/CD147 of transfected cells selected by G418 (geneticin) was observed by immuno-histochemical SP staining and FACS (fluorescence activated cell sorting). Gelatin zymography was used to determine the effect of PCI-asHAb18G on reducing secretions of MMP-2 and MMP-9 of the transfected cells. Boyden chamber was employed to test the invasion of HCC cells in vitro. RESULTS: The construction of antisense RNA vector PCI-asHAb18G was verified correct by partial nucleotide sequencing and restricted endonuclease digestion. The expression of HAb18G/CD147 in transfected HHCC was inhibited by PCI-asHAb18G. Secretions of MMP-2 and MMP-9 of transfected HHCC were reduced and the invasion of transfected HHCC was inhibited compared to HHCC, respectively. CONCLUSION: Invasion of HCC cells can be inhibited by antisense RNA of HAb18G/CD147. HAb18G/CD147 may be used as a potential target of drugs for anti-invasion and metastasis of HCC.

Antigens, CD↗

Experimental study of anti-tumor effects of polysaccharides from Angelica sinensis.

AIM: To investigate the in vivo anti-tumor effects of total polysaccharide (AP-0) isolated from Angelica sinensis (Oliv.) Diels (Danggui) on mice and the in vitro inhibitory effects of AP-0 and the sub-constituents (AP-1, AP-2 and AP-3) separated from AP-0 on invasion and metastasis of human hepatocellular carcinoma. METHODS: Three kinds of murine tumor models in vivo, sarcoma 180 (S180), leukemia L1210 and Ehrlich ascitic cancer (EAC) were employed to investigate the anti-tumor effects of AP-0. For each kind of tumor model, three experimental groups were respectively given AP-0 at doses of 30, 100 and 300 mg/kg by ip once a day for 10 days. Positive control groups were respectively given Cy at a dose of 30 mg/kg for S180 and leukemia L1210, and 5-FU at a dose of 20 mg/kg for EAC. On d 11, mice bearing S180 were sacrificed and the masses of tumors, spleens and thymus weighed. The average living days of mice bearing EAC and of mice bearing L1210 were observed, and the rates of life prolongation of each treatment were calculated, respectively. The inhibitory effects of APs on hepatoma invasion and metastasis in vitro were investigated by employing human hepatocellular carcinoma cell line (HHCC) with the Matrigel invasion chamber, adhesion to extracellular matrix and chemotatic migration tests, respectively. RESULTS: AP-0 had no obviously inhibitory effect on the growth of S180, but it could significantly decrease the thymus weights of the mice bearing S180. AP-0 could significantly reduce the production of ascitic liquids and prolong the life of mice bearing EAC. AP-0 could also increase the survival time of mice bearing L1210. AP-0 and AP-2 had significantly inhibitory effects on the invasion of HHCC into the Matrigel reconstituted basement membrane with the inhibitory rates of 56.4 % and 68.3 %, respectively. AP-0, AP-1, AP-2 and AP-3 could influence the adhesion of HHCC to extracellular matrix proteins (Matrigel and fibronectin) at different degrees, among them only AP-3 had significant blocking effect on the adhesion of HHCC to fibronectin with an inhibitory rate of 30.3 %. AP-0, AP-1 and AP-3 could partially inhibit the chemotactic migration abilities of HHCC. CONCLUSION: The experimental findings suggest that the total polysaccharide of Angelica sinensis (Oliv.) Diels (Chinese Danggui) possesses anti-tumor effects on experimental tumor models in vivo and inhibitory effects on invasion and metastasis of hepatocellular carcinoma cells in vitro.

Angelica sinensis↗

[Bio-panning of antagonistic peptides against HAb18G/CD147 and their function of anti-hepatoma invasion].

OBJECTIVE: To screen out the HAb18G/CD147 binding peptides and find out an antagonist against hepatoma invasion. METHODS: HAb18G/CD147 was purified by affinity chromatographic method and the antigen binding peptides acquired by bio-panning a phage-displayed 12-peptide library. After obtaining the sequence of the selected phage-displayed peptides, all the 9 peptides were synthesized by solid-phase method and identified by mass spectrograph. The peptides' anti-metastatic function was tested by Boyden Chamber assay. RESULTS: The purified HAb18G/CD147, identified by Western blot (molecular weight about 65 kd) could be used to bio-pan the phage-displayed peptide library. After 3 rounds of bio-panning, 9 positive phage clones were selected and sequenced. The synthesized peptides had uneven inhibitory activities and three of them were able to markedly inhibit the hepatoma cell invasion (P < 0.01). The most effective peptide decreased by 90.1% of hepatoma cells migrating through the Boyden Chamber membrane as compared with the control. CONCLUSION: Bio-panning the phage-displayed peptide library can be used successfully to screen out the antigen binding peptides. Hepatoma metastatic potential can be inhibited by peptide antagonist which could be a good foundation of developing peptide therapeutic agent against hepatoma metastasis.

Animals↗

[Study on the affinities of antagonistic peptides with HAb18G/CD147 on the hepatocellular carcinoma cells].

AIM: To study the affinities of antagonistic peptides with HAb18G/CD147 on the hepatocellular carcinoma cells. METHODS: Fluorescence-activated cell sorting analysis(FACS) was used to detect the expression of HAb18G antigen on HHCC and SMMC7721 cells. Antagonistic peptides(APs) AP-1, AP-2 and AP-6 were labelled with biotin at their N-terminals. The binding abilities of APs to HHCC or SMMC7721 cells were determined by FACS and confocal microscope. RESULTS: HAb18G antigen was highly expressed on the HHCC and SMMC7721 cells. AP-6 has the highest affinity for HHCC and SMMC7721 cells compared with AP-1 and AP-2. CONCLUSION: Antagonistic peptides of HAb18G/CD147 have high affinities with HAb18G/CD147 on the hepatocellular carcinoma cells.

Basigin↗

[Multiagent system for individual artificial hip joint].

Based on the introduction of multi-agent and individual artificial hip joints, this paper reports a multi-agent design system for individual artificial hip joints. Three agents are defined, the first is to optimize the diameter of hip joint, the second for FEM, the third for simulation between the prosthesis and acetabulum cup. The paper lays emphasis on the second agent.

Acetabulum↗

[Effects of Angelica polysaccharide on blood coagulation and platelet aggregation].

OBJECTIVE: To investigate the effects of angelica polysaccharide (AP) on blood coagulation and platelet aggregation. METHODS: Infrared turbidimetric method was used to estimate platelet aggregation, active partial thromboplastin time (APTT), prothrombin time (PT) and thrombin time (TT); bleeding time (BT) was measured by cutting the mouse's tail and coagulate time (CT) was measured by dropping the blood on carry sheet glass. RESULTS: AP prominently enhanced the platelet aggregation at 5 min, while showed less effects on the maximum platelet aggregation. It also markedly prolonged CT but shortened BT. AP significantly prolonged TT and APTT in dosage of 3 mg/kg and 10 mg/kg, while showed no obvious effect on PT. CONCLUSION: These results suggest that AP has potent anticoagulant and haemostasis effects. The haemostasis effect is related to promoting platelet aggregation.

Angelica sinensis↗