OBJECTIVE: Diabetes-induced gastrointestinal (GI) motility disorders are increasingly prevalent. Damage to the enteric nervous system (ENS), composed primarily of enteric neurons and glial cells, is an essential mechanism involved in these disorders. Although electroacupuncture (EA) has shown the potential to mitigate enteric neuronal loss, its mechanism is not fully understood. Additionally, the effects of EA on enteric glial cells have not been investigated. Enteric neural precursor cells (ENPCs) contribute to the structural and functional integrity of the ENS, yet whether EA enhances their differentiation into enteric neurons and glial cells remains unexplored. This study investigates whether EA promotes ENS repair through enhancing ENPC-derived neurogenesis and gliogenesis and elucidates the potential molecular mechanisms involved. METHODS: Transgenic mice were used to trace Nestin⁺/nerve growth factor receptor (Ngfr)⁺ ENPCs labeled with green fluorescent protein (GFP) in vivo. Mice were randomly divided into four groups: control, diabetes mellitus (DM), DM + sham EA, and DM + EA. The effects of EA on diabetic mice were evaluated by GI motility, ENS structure, and ENPC differentiation. Glial cell line-derived neurotrophic factor (GDNF)/Ret signaling was detected to clarify the underlying molecular mechanisms. RESULTS: EA alleviated diabetes-induced GI motility disorders, as indicated by reduced whole gut transit time, shortened colonic bead expulsion time, and enhanced smooth muscle contractility. Furthermore, EA attenuated diabetes-induced losses of enteric neurons and glial cells, thereby restoring ENS integrity. Notably, EA reversed the diabetes-induced decrease in ENPCs and significantly increased the absolute number and the proportion of ENPC-derived enteric neurons. However, immunofluorescence analyses revealed no colocalization between EA-induced glial fibrillary acidic protein⁺ glial cells and GFP-labeled ENPCs. Mechanistically, GDNF/Ret signaling was elevated in intestinal tissues and upregulated in ENPCs in EA-treated diabetic mice. CONCLUSION EA facilitates ENS repair by promoting Nestin⁺/Ngfr⁺ ENPC differentiation into enteric neurons via upregulation of GDNF/Ret signaling, and driving enteric gliogenesis from non-Nestin⁺/Ngfr⁺ ENPCs. These findings highlight EA's role in ameliorating diabetes-induced GI dysmotility through ENPC-derived ENS restoration. Please cite this article as: Guo JL, Liu S, Ding SJ, Yang X, Du F. Electroacupuncture at ST36 improves gastrointestinal motility disorders by promoting enteric nervous system regeneration through GDNF/Ret signaling in diabetic mice. J Integr Med. 2025; 23(5):548-559.
Animals ; Electroacupuncture ; Enteric Nervous System/physiology* ; Gastrointestinal Motility/physiology* ; Glial Cell Line-Derived Neurotrophic Factor/metabolism* ; Diabetes Mellitus, Experimental/therapy* ; Signal Transduction ; Mice ; Gastrointestinal Diseases/physiopathology* ; Proto-Oncogene Proteins c-ret/metabolism* ; Mice, Transgenic ; Male ; Nerve Regeneration ; Neural Stem Cells ; Mice, Inbred C57BL ; Acupuncture Points

Objective: To investigate the clinicopathological features, treatment and prognosis of patients with RET fusion positive non-small cell lung cancer (NSCLC). Methods: A total of 1 089 NSCLCs were retrieved at Affiliated Hospital of Jiangnan University from August 2018 to April 2020. In all cases, multiple gene fusion detection kits (fluorescent PCR method) were used to detect the gene status of RET, EGFR, ALK, ROS1, KRAS, BRAF and HER2; and immunohistochemical method was used to detect the expression of PD-L1 and mismatch repair related proteins. The correlation between RET-fusion and patients' age, gender, smoking history, tumor stage, grade, pathologic type, and PD-L1, mismatch repair related protein expression was analyzed. Results: There were 22 cases (2.02%) detected with RET fusion-positive in 1 089 NSCLC patients, in which 11 males and 11 females; and the median age was 63.5 years. There were 20 adenocarcinomas, including 11 acinar predominant adenocarcinoma (APA), five solid predominant adenocarcinoma (SPA) and four lepidic predominant adenocarcinoma (LPA); There were one case each of squamous cell carcinoma (non-keratinizing type) and sarcomatoid carcinoma (pleomorphic carcinoma). There were 6 and 16 patients with RET fusion-positive who were in stage Ⅰ-Ⅱ and Ⅲ-Ⅳ respectively, and 16 cases with lymph node metastasis, 11 cases with distant metastasis. Among RET fusion-positive cases, one was detected with HER2 co-mutation. The tumor proportion score of PD-L1≥1% in patients with RET fusion positive lung cancer was 54.5% (12/22). Defects in mismatch repair protein expression were not found in patients with RET fusion positive NSCLC. Four patients with RET fusions positive (two cases of APA and two cases of SPA) received pratinib-targeted therapy, and two showed benefits from this targeted therapy. Conclusions: The histological subtypes of RET fusions positive NSCLC are more likely to be APA or SPA. RET fusion-positive NSCLC patients are associated with advanced clinical stage, lymph node metastases, and they may benefit from targeted therapy with RET-specific inhibitors.
Male ; Female ; Humans ; Middle Aged ; Carcinoma, Non-Small-Cell Lung/pathology* ; Lung Neoplasms/pathology* ; B7-H1 Antigen/genetics* ; Protein-Tyrosine Kinases/genetics* ; Proto-Oncogene Proteins c-ret/metabolism* ; Proto-Oncogene Proteins/genetics* ; Adenocarcinoma/pathology* ; Carcinoma, Squamous Cell/genetics* ; Mutation

PURPOSE: Rearrangement of the proto-oncogene rearranged during transfection (RET) has been newly identified potential driver mutation in lung adenocarcinoma. Clinically available tyrosine kinase inhibitors (TKIs) target RET kinase activity, which suggests that patients with RET fusion genes may be treatable with a kinase inhibitor. Nevertheless, the mechanisms of resistance to these agents remain largely unknown. Thus, the present study aimed to determine whether epidermal growth factor (EGF) and hepatocyte growth factor (HGF) trigger RET inhibitor resistance in LC-2/ad cells with CCDC6-RET fusion genes. MATERIALS AND METHODS: The effects of EGF and HGF on the susceptibility of a CCDC6-RET lung cancer cell line to RET inhibitors (sunitinib, E7080, vandetanib, and sorafenib) were examined. RESULTS: CCDC6-RET lung cancer cells were highly sensitive to RET inhibitors. EGF activated epidermal growth factor receptor (EGFR) and triggered resistance to sunitinib, E7080, vandetanib, and sorafenib by transducing bypass survival signaling through ERK and AKT. Reversible EGFR-TKI (gefitinib) resensitized cancer cells to RET inhibitors, even in the presence of EGF. Endothelial cells, which are known to produce EGF, decreased the sensitivity of CCDC6-RET lung cancer cells to RET inhibitors, an effect that was inhibited by EGFR small interfering RNA (siRNA), anti-EGFR antibody (cetuximab), and EGFR-TKI (Iressa). HGF had relatively little effect on the sensitivity to RET inhibitors. CONCLUSION: EGF could trigger resistance to RET inhibition in CCDC6-RET lung cancer cells, and endothelial cells may confer resistance to RET inhibitors by EGF. E7080 and other RET inhibitors may provide therapeutic benefits in the treatment of RET-positive lung cancer patients.
Adenocarcinoma/drug therapy/*genetics ; Cell Line, Tumor ; Cetuximab/pharmacology ; Drug Resistance, Neoplasm/drug effects/*genetics ; Epidermal Growth Factor/metabolism/*pharmacology ; *Gene Rearrangement ; Hepatocyte Growth Factor/*pharmacology ; Humans ; Indoles/pharmacology ; Lung Neoplasms/drug therapy/*genetics ; MAP Kinase Signaling System ; *Mutation ; Niacinamide/analogs & derivatives/pharmacology ; Phenylurea Compounds/pharmacology ; Piperidines/pharmacology ; Protein Kinase Inhibitors/therapeutic use ; Proto-Oncogene Proteins c-ret/*antagonists & inhibitors/genetics ; Pyrroles/pharmacology ; Quinazolines/pharmacology ; RNA, Small Interfering/pharmacology ; Receptor, Epidermal Growth Factor/genetics/metabolism ; Signal Transduction/drug effects ; fms-Like Tyrosine Kinase 3/metabolism

OBJECTIVETo explore the neuroprotective effects of electroacupuncture (EA) on hypoxic-ischemic encephalopathy (HIE) and to further investigate the role of glial cell line-derived neurotrophic factor (GDNF) family receptor member RET (rearranged during transfection) and its key downstream phosphatidylinositol 3 kinase (PI-3K)/protein kinase B (Akt) pathway in the process.

METHODSA total of 220 seven-day-old SD rats (of either sex, from 22 broods) were randomly divided into two groups, one (30 rats) for sham-surgery group and the other (190 rats) for HIE model group. The HIE model was established using the left common carotid artery ligation method in combination with hypoxic treatment. The successfully established rats were randomly divided into five groups, including control model group, EA group, sham-EA group, antagonist group and antagonist plus electroacupuncture group, with 35 rats in each group. Baihui (GV 20), Dazhui (GV 14), Quchi (LI 11) and Yongquan (KI 1) acupoints were chosen for acupuncture. EA was performed at Baihui and Quchi for 10 min once a day for continuous 1, 3, 7 and 21 days, respectively. The rats were then killed after the operation and injured cerebral cortex was taken for the measurement of neurologic damage by hematoxylin-eosin (HE) staining and the degenerative changes of cortical ultrastructure by transmission electron microscopy. RET mRNA level and Akt protein level were detected by real-time reverse-transcription polymerase chain reaction (RT-PCR) and western blot analysis, respectively.

RESULTSEA could ameliorate neurologic damage of the first somatic sensory area (S1Tr) and alleviate the degenerative changes of ultrastructure of cortical neurons in rats subjected to HIE. And the longer acupuncture treatment lasted, the better its therapeutic effect would be. This was accompanied by gradually increased expression of GDNF family receptor RET at the mRNA level and its downstream signaling Akt at the protein level in the ischemic cortex.

CONCLUSIONEA has neuroprotective effects on HIE and could be a potential therapeutic strategy for HIE in the neonate. Activation of RET/Akt signaling pathway might be involved in this process.

Animals ; Blotting, Western ; Cerebral Cortex ; pathology ; ultrastructure ; Electroacupuncture ; Female ; Glial Cell Line-Derived Neurotrophic Factor ; genetics ; metabolism ; Hypoxia-Ischemia, Brain ; genetics ; pathology ; therapy ; Male ; Nerve Degeneration ; pathology ; Neurons ; pathology ; ultrastructure ; Neuroprotective Agents ; therapeutic use ; Proto-Oncogene Proteins c-akt ; genetics ; metabolism ; Proto-Oncogene Proteins c-ret ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Rats, Sprague-Dawley ; Real-Time Polymerase Chain Reaction

OBJECTIVETo study the prevalence of ALK, ROS1 and RET fusion genes in non-small cell lung cancer (NSCLC), and its correlation with clinicopathologic features.

METHODSFormalin-fixed and paraffin-embedded tissue sections from samples of 302 patients with NSCLC were screened for ALK, ROS1, RET fusions by real-time polymerase chain reaction (PCR). All of the cases were validated by Sanger DNA sequencing. The relationship between ALK, ROS1, RET fusion genes and clinicopathologic features were analyzed.

RESULTSIn the cohort of 302 NSCLC samples, 3.97% (12/302) were found to contain ALK fusion genes, including 3 cases with E13; A20 gene fusion, 3 cases with E6; A20 gene fusion and 3 cases with E20; A20 gene fusion. There was no statistically significant difference in patient's gender, age, smoking history and histologic type. Moreover, in the 302 NSCLC samples studied, 3.97% (12/302) were found to contain ROS1 fusion genes, with CD74-ROS1 fusion identified in 9 cases. There was no statistically significant difference in patients' gender, age, smoking history and histologic type. One non-smoking elderly female patient with pulmonary adenocarcinoma had RET gene fusion. None of the cases studied had concurrent ALK, ROS1 and RET mutations.

CONCLUSIONSThe ALK, ROS1 and RET fusion gene mutation rates in NSCLC are low, they represent some specific molecular subtypes of NSCLC. Genetic testing has significant meaning to guide clinical targeted therapy.

Adenocarcinoma ; Aged ; Carcinoma, Non-Small-Cell Lung ; genetics ; metabolism ; Female ; Gene Fusion ; Genetic Testing ; Humans ; Lung Neoplasms ; Mutation ; Oncogene Proteins, Fusion ; genetics ; metabolism ; Protein-Tyrosine Kinases ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-ret ; genetics ; metabolism ; Real-Time Polymerase Chain Reaction ; Receptor Protein-Tyrosine Kinases ; genetics ; metabolism ; Sequence Analysis, DNA ; Smoking

The phosphoinositide 3-kinase-AKT-mammalian target of rapamycin (PI3K-AKT-mTOR) pathway is a frequently hyperactivated pathway in cancer and is important for tumor cell growth and survival. The development of targeted therapies against mTOR, a vital substrate along this pathway, led to the approval of allosteric inhibitors, including everolimus and temsirolimus, for the treatment of breast, renal, and pancreatic cancers. However, the suboptimal duration of response in unselected patients remains an unresolved issue. Numerous novel therapies against critical nodes of this pathway are therefore being actively investigated in the clinic in multiple tumor types. In this review, we focus on the progress of these agents in clinical development along with their biological rationale, the need of predictive biomarkers and various combination strategies, which will be useful in counteracting the mechanisms of resistance to this class of drugs.
Antineoplastic Agents ; therapeutic use ; Drug Resistance, Neoplasm ; Humans ; Molecular Targeted Therapy ; Neoplasms ; drug therapy ; genetics ; metabolism ; PTEN Phosphohydrolase ; genetics ; metabolism ; Phosphatidylinositol 3-Kinases ; antagonists & inhibitors ; metabolism ; Proto-Oncogene Proteins c-akt ; antagonists & inhibitors ; metabolism ; Proto-Oncogene Proteins c-ret ; metabolism ; Sirolimus ; therapeutic use ; TOR Serine-Threonine Kinases ; antagonists & inhibitors ; metabolism

OBJECTIVETo evaluate the expressions of wildtype-RET (WT-RET) and RET/PTC in sporadic adult papillary thyroid carcinoma and to investigate their clinicopathologic correlation.

METHODSSixty-six papillary thyroid carcinomas (PTC) and thirty-six control cases with frozen and paraffin-embedded tissues were analyzed for the expressions of WT-RET and oncogene RET/PTC1 or RET/PTC3 by nested RT-PCR.

RESULTS(1) 62 percent (41/66) of PTC patients were above 40 years of age. Thirty-eight percent (25/66) of the tumors showed lymphocytic thyroiditis. Lymph node and distant metastasis were seen in 59% (39/66) and 7.6% (5/66) respectively. (2) Forty-five cases (68.1%) of PTCs expressed RET tyrosine kinase domain (RET-TK). Simultaneous expressions of RET-BP and TK were seen in nineteen PTCs (28.8 %). One of eight adenomas (12.5 %) expressed wild-type RET (WT-RET). (3) Fourteen PTCs (21.2%) expressed RET/PTC, including five cases expressing RET/PTC1 and nine cases expressing RET/PTC3. Six cases (9%) expressed both RET/PTC and WT-RET. (4) Statistic analysis did not show any correlation between the expression of WT-RET or RET/PTC and clinicopathologic parameters.

CONCLUSIONSThe expression of RET/PTC was specific to PTC. However, its prevalence was low and, therefore, of limited diagnostic utility. The expression patterns of WT-RET in PTC and adenoma suggest that there are different molecular mechanisms in activating RET proto-oncogene in thyroid tumors.

Adenoma ; genetics ; metabolism ; Adolescent ; Adult ; Aged ; Carcinoma, Papillary ; genetics ; metabolism ; pathology ; Female ; Gene Rearrangement ; Hashimoto Disease ; metabolism ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Oncogene Proteins, Fusion ; biosynthesis ; genetics ; Protein-Tyrosine Kinases ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-ret ; biosynthesis ; genetics ; RNA, Messenger ; biosynthesis ; genetics ; Thyroid Neoplasms ; genetics ; metabolism ; pathology

OBJECTIVEHirschsprung's Disease (HD) and allied Hirschsprung's disorder (HAD) have very similar clinical manifestations, but there are many different theories about the two diseases. The present study was designed to understand the expression of Ret protein in HD and HAD, and to explore the role of Ret protein in the pathogenesis of HD and HAD.

METHODSColon specimens from patients with confirmed HD and HAD, including 15 cases of HD (male 12, female 3) and 11 cases of HAD (male 8, female 3) were collected for this study. At the same time normal colon specimens from 10 individuals with other diseases were used as control. The expression of Ret protein in the intestinal tissue was detected by using immunohistochemical SABC technique with mouse anti-Ret monoclonal antibody.

RESULTSIn the colon specimens from normal controls and the dilated segments of colon from HD and HAD patients, moderate to large number of Ret-positive cells were observed among the ganglion cells of myenteric plexuses and submucosal plexuses (P > 0.05). On the contrary, Ret-positive cells were not seen in the stenotic segment of colon from HD patients. But there was positive staining in the stenotic segment of the colon from HAD. Moreover, giant ganglion cells showing strongly positive staining could be seen. There were also displastic cells, small cells, and cells with irregular shape. Statistical analysis showed significant differences in Ret cells positivity between the stenotic segment of colon of HD and the normal control (P < 0.001) as well as between HD and HAD (P < 0.001).

CONCLUSIONRet protein may play an important role in the pathogenesis of HD and could not have definite relationship with HAD.

Case-Control Studies ; Child ; Child, Preschool ; Colon ; pathology ; Female ; Hirschsprung Disease ; genetics ; metabolism ; Humans ; Infant ; Male ; Neurons ; pathology ; Proto-Oncogene Proteins c-ret ; genetics ; metabolism

OBJECTIVETo investigate the expression of cytokeratins and ret in thyroid papillary carcinoma (TPC) and their diagnostic value.

METHODSDuring the period of October 1999 to March 2002, 69 cases of TPC (42 cases with adjacent normal thyroid tissue) and 14 cases of nodular goiter with papillary hyperplasia were enrolled into the study. Immunohistochemistry for CK19, CK17, CK8, CK20 and ret was performed in all cases using EnVision and LSAB methods respectively.

RESULTSThe positive rates for CK19 and ret in TPCs were 85.5% and 68.1% respectively, which were significantly (P < 0.01) higher than those in nodular goiter and normal thyroid tissue (25.0% and 5.4% respectively). The expression of CK17 was also observed in a few cases of TPCs (11/69, 15.9%), which was mainly localized in areas of squamous metaplasia, poorly differentiated carcinoma and/or in the small infiltrative foci. The positive rates for CK8 were 75.4% and 26.8% in TPCs and benign thyroid tissue respectively. All cases were negative for CK20.

CONCLUSIONSCK19, CK17 and ret expressions are significantly higher in TPCs than benign thyroid tissue; and this characteristic can have important diagnostic value.

Carcinoma, Papillary ; metabolism ; pathology ; Humans ; Immunohistochemistry ; Keratins ; analysis ; biosynthesis ; Proto-Oncogene Proteins ; biosynthesis ; Proto-Oncogene Proteins c-ret ; Receptor Protein-Tyrosine Kinases ; biosynthesis ; Thyroid Neoplasms ; metabolism ; pathology