1.Decreased expression of β-nerve growth factor correlated with histological changes in a cryptorchidism rat model.
Hua XIAN ; Yun XIAN ; Chun-yi JIANG ; Xiao NIE ; Xu-dong WANG ; Hong-xia CHENG ; Jiang-hong HE ; Yong-jun WANG ; Yan ZHOU ; Jian-fei HUANG
Chinese Medical Journal 2012;125(4):713-716
BACKGROUNDNerve growth factor (NGF) is well-known for its important role in the development and maintenance of the nervous system. Along with its neurotrophic role, NGF has been detected in the testis of mouse, rat and human, suggesting an additional non-neurotrophic effect in the male reproductive system. The expression of β-NGF in the undescended testes (cryptorchidism) has not been detected at present. The aim of this study was to evaluate the expression of β-nerve growth factor mRNA and protein in experimental cryptorchidism.
METHODSA unilateral mechanical cryptorchidism model in the Sprague-Dawley rat was established and the expression of β-NGF with histologic changes in experimental cryptorchidism were investigated using one step quantitative real-time reverse transcription-polymerase chain reaction, in situ hybridization histochemistry, immunofluorescence and hematoxylin-eosin staining.
RESULTSThe expression of β-NGF mRNA and protein were both significantly decreased in the development of unmarred testis and cryptorchidism-induced testis, and the decrease of β-NGF in cryptorchidism-induced testis was far greater than that in uninjured testis.
CONCLUSIONFrom this investigation, we confirmed a lower expression of β-NGF in undescended testes than in the development of testis.
Animals ; Cryptorchidism ; genetics ; metabolism ; Male ; Nerve Growth Factor ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley
2.Study of neurotrophic factor receptor tyrosine kinase gene fusion in the precise treatment of wild-type gastrointestinal stromal tumor.
Hai Dong ZHANG ; Xiao Nan YIN ; Zhao Lun CAI ; Bo ZHANG
Chinese Journal of Gastrointestinal Surgery 2021;24(9):769-774
The neurotrophin receptor kinase (NTRK) gene encodes neurotrophic factor receptor tyrosine kinase (NTRK), which plays an important role in the development and function of the nervous system. NTRK gene fusion mutation results in the production of chimeric NTRK proteins, which have carcinogenic potential through constitutive activation or overexpression. NTRK gene fusion mutation can lead to a special type of wild type gastrointestinal stromal tumor (GIST), whose clinical manifestations and treatment are completely different from other types of GIST. This fusion mutation can be detected clinically by a variety of methods, including tumor DNA and RNA sequencing and immunohistochemical staining. In patients with NTRK fusion positive tumors, NTRK inhibitors such as larotrectinib and entrectinib have shown good antitumor efficacy, with clinical response rates as high as 75%. Therefore, there is a need to improve the recognition and detection of fuch patients and to improve their prognosis by individualized and precise treatment with TRK inhibitors.
Gastrointestinal Stromal Tumors/genetics*
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Gene Fusion
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Humans
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Neoplasms
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Nerve Growth Factors
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Protein Kinase Inhibitors
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Receptor, trkA/genetics*
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Receptors, Nerve Growth Factor/genetics*
3.Molecular genetics of Kallmann syndrome: an update.
Chao FU ; Zheng FENG ; Rui-Zhi LIU
National Journal of Andrology 2011;17(4):361-365
Kallmann syndrome (KS) is a clinically and genetically heterogeneous disorder that occurs in either an inherited or a sporadic manner. KS results from failed embryonic migration of GnRH-1 neurons from the nasal placode to the hypothalamus, due to the abnormal development of olfactory nerves and bulbs. Hypogonadotropic hypogonadism is related to GnRH deficiency, and anosmia is associated with the absence or hypoplasia of olfactory bulbs and tracts. KS patients can also present some non-reproductive or non-olfactory anomalies in addition to the above typical symptoms. For the high complexity of the molecular genetic mechanism of KS, to date, only 6 KS-related genes have been identified. The KAL1 gene is responsible for the X chromosome-linked recessive form of KS, while the fibroblast growth factor receptor 1 (FGFR1/KAL2) and fibroblast growth factor 8 (FGF8/KAL6) genes are related to the autosomal dominant form of the disease. However, the mutations in these 6 genes account for only about 25 - 30% of all KS cases, which suggests that other pathogenic genes involved in KS remain to be discovered. This article presents an overview on the studies of the pathogenic genes, clinical diagnosis and treatment of KS.
Extracellular Matrix Proteins
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genetics
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Humans
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Kallmann Syndrome
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genetics
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Mutation
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Nerve Tissue Proteins
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genetics
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Receptor, Fibroblast Growth Factor, Type 1
;
genetics
4.Effect of HSV-1 infection on NGF and its receptor expression in human glioma cells.
Yun HOU ; Ling LI ; Ming HU ; Guang-Yu JIANG ; Qian WANG ; Dong-Meng QIAN ; Zhi-Yong YAN ; Wei ZHAO ; Xu-Xia SONG ; Bin WANG
Chinese Journal of Virology 2010;26(6):477-482
Nerve growth factor (NGF) is mainly secreted by the neuroglia cells, which can exert biological effect through its receptors on the specific target cell surface. NGF is closely related to neurocyte growth, differentiation and apoptosis. As a neurotropic virus, HSV-1 an easily lead to neurocyte, neuroglia cells death or apoptosis. In this study, the U251 human glioma cells were chosen as target cells to study the change of NGF and its receptors in the apoptosis process of HSV-1 infection. Our results showed that U251 cells were permissive to HSV-1 replication. In the apoptosis process of HSV-1 infected U251 cells, the expression of both NGF and P75NTR increased and then decreased, while the expression of TrkA decreased gradually. These result indicated that HSV-1 was able to induce the abnormal expression of NGF and its receptors in U251 cells.
Apoptosis
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Cell Line, Tumor
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Gene Expression
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Glioma
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genetics
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metabolism
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physiopathology
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virology
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Herpes Simplex
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genetics
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metabolism
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physiopathology
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virology
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Herpesvirus 1, Human
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genetics
;
physiology
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Humans
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Nerve Growth Factor
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genetics
;
metabolism
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Receptor, Nerve Growth Factor
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genetics
;
metabolism
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Receptor, trkA
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genetics
;
metabolism
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Virus Replication
5.Expression of human nerve growth factor beta gene in central nervous system mediated by recombinant adeno-associated viruses type-2 vector.
Kai GAO ; Yong-jie WU ; Xiao-bing WU ; Chun-ming RAO ; Jun-zhi WANG
Chinese Medical Journal 2004;117(9):1370-1376
BACKGROUNDNeurone atrophy and loss are major causes of chronic neurodegenerative disorders such as Alzheimer's disease. Despite many pharmacotherapies for neurodegeneration, there are no accepted treatments. We investigated the feasibility of human nerve growth factor beta (hNGFbeta) gene expression mediated by recombinant adeno-associated viruses type-2 (rAAV-2) vector in the central nervous system (CNS) after blood brain barrier (BBB) disruption.
METHODSrAAV-2 containing hNGFbeta gene was constructed. The ability of hNGFbeta gene mediated by rAAV-2 vector (rAAV-2/hNGFbeta) to transfect cells in vitro was confirmed by both ELISA and bioassay of hNGFbeta in the culture supernatant of BHK-21 cells infected by rAAV-2/hNGFbeta. rAAV-2/hNGFbeta and rAAV-2/green fluorescence protein (GFP) were administrated separately to rat brains through internal carotid intubation after BBB disruption with hypertonic mannitol. Brain hNGFbeta concentration was measured by ELISA and GFP in brain sections was examined by laser scan confocal microscope.
RESULTSAfter 48 hours, hNGFbeta content in supernatant was up to (188.0 +/- 28.6) pg/ml when BHK-21 cells were infected by rAAV-2/hNGFbeta at multiplicity of infection (MOI) 1.0 x 10(6) vector genome. Neurone fibre outgrowths were obvious in dorsal root ganglion neurone assays by adding serum free culture medium harvested from BHK-21 cells exposed to rAAV-2/hNGFbeta. Whole brain hNGFbeta content in rAAV-2/hNGFbeta transferred group was up to (636.2 +/- 140.6) pg/ml. hNGFbeta content of BBB disruption in rAAV-2/hNGFbeta infused group increased significantly compared to the control group (P <0.05). GFP expression was clearly observed in brain sections of rAAV-2/GFP transferred group.
CONCLUSIONrAAV-2/hNGFbeta successfully expresses in the CNS after BBB disruption induced by hypertonic mannitol.
Animals ; Blood-Brain Barrier ; Brain ; metabolism ; Cricetinae ; Dependovirus ; genetics ; Genetic Vectors ; genetics ; Humans ; Nerve Growth Factor ; genetics ; Recombination, Genetic
6.Progress of gene therapy for erectile dysfunction.
Yun CHEN ; Yu-tian DAI ; Ze-yu SUN
National Journal of Andrology 2006;12(11):1016-1020
Gene therapy is currently investigated in animal studies for treating erectile dysfunction (ED), and is affording an conspicuous therapeutic possibility for the treatment of ED, especially in L-arg-NO-cGMP pathway, ion channel, the protection of nerves and endothelia in erectile tissues. However there still exist so many problems for gene therapy to be effectively applied to the clinical treatment of ED. This review aims to examine the experimental efforts in recent years and tries to give a brief introduction to the new approaches in the field of ED researches.
Animals
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Cell Communication
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genetics
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Erectile Dysfunction
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therapy
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Genetic Therapy
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Ion Channels
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genetics
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Male
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Nerve Growth Factors
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genetics
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Rats
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Transfection
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Vascular Endothelial Growth Factor A
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genetics
7.Effects of transforming growth factor-beta 1 on the peripheral nerve regeneration of rats.
Yuan-yuan PEI ; Shao-bin DUAN ; Wei-jun CAI ; Xi-nan YI ; Zhi-cheng ZENG ; Jian-wei ZHANG ; Yuan-zhong XU ; Qiong-yan ZOU ; Xiao-dan WEN
Journal of Central South University(Medical Sciences) 2005;30(4):447-451
OBJECTIVE:
To explore the effects of exogenous transforming growth factor-beta 1 (TGFbeta1) on peripheral nerve regeneration after the peripheral nerve injury and if TGFbeta1 regulates the expression of basic fibroblast growth factor (bFGF) in the anterior horn motoneurons of spinal cord during regeneration.
METHODS:
Forty-eight rats were crushed on the right sciatic nerve and then randomly divided into 2 groups: TGFbeta1 group and NS group. In TGFbeta1 group, TGFbeta1 50 microL (0.1 microg/mL) was injected into the proximal nerve near to the crushed nerve and after the operation the injured leg was injected with equal TGFbeta1 whereas the NS was replaced in the NS group. The rats of each group survived for 3, 7, 14 and 21 days after the lesion. The bFGF expression in the anterior horn motoneurons of spinal cord was detected by immunohistochemistry (IHC). Semi-thin section and Fast Blue retrograde tracing were also performed with the rats surviving for 21 days to observe the regeneration of distal end in the injured right sciatic nerve.
RESULTS:
The number of bFGF immunoreactive positive motoneurons in TGFbeta1 group was obviously higher than that of the NS group (P < 0.05). In the distal sciatic nerve of the rats treated with TGFbeta1, the number and diameter of regenerating myelinated axons and the thickness of myelinated sheath were more than those of the NS group (P < 0.05). The number of motoneurons in spinal cord and neurons in dorsol root ganglia (DRG) labelled with Fast Blue in the NS group was obviously lower than in the TGFbeta1 group (P < 0.01).
CONCLUSION
Exogenous TGFbeta1 plays an important role in promoting the peripheral nerve regeneration; TGFbeta1 up-regulates the bFGF expression in the anterior horn motoneurons of spinal cord during the peripheral nerve regeneration.
Animals
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Female
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Fibroblast Growth Factor 2
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biosynthesis
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genetics
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Male
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Motor Neurons
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metabolism
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Nerve Regeneration
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drug effects
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Random Allocation
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Rats
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Rats, Sprague-Dawley
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Sciatic Nerve
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injuries
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metabolism
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physiology
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Spinal Cord
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metabolism
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Transforming Growth Factor beta
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pharmacology
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Transforming Growth Factor beta1
8.Expression, purification and renaturation of proNGF in Escherichia coli.
Hanmin JIANG ; Xinjun CHAI ; Bing HE ; Juan ZHAO ; Xinda YU
Chinese Journal of Biotechnology 2008;24(3):509-514
Nerve growth factor (NGF) promotes neuronal survival and differentiation and stimulates neurite outgrowth. NGF is synthesized as a precursor-proNGF in vivo. In this paper, a pET-proNGF prokaryocyte expression vector was constructed and transformed into E. coli BL21(DE3)pLysS. The proNGF was expressed in the form of non-active aggregated monomer in E. coli after induction with IPTG. SDS-PAGE revealed the proNGF expression product had a Mr.30.2 kD. Western blotting analysis showed that the protein had good antigenicity. Fusion protein was successfully purified by Ni2+-NTA affinity chromatography and cleaved by Enterokinase and 13.1 mg proNGF was obtained from 100 mL cell culture in a typical experiment. The protein was dialyzed in a redox system containing reduced and oxidized glutathione. RP-HPLC was used to analysis the result of the refolding. The refolded proNGF protein can induce neurite outgrowth of PC12 cells, which indicated that pro-form of NGF we obtained had biological activity.
Escherichia coli
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genetics
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metabolism
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Genetic Vectors
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genetics
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Humans
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Nerve Growth Factor
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biosynthesis
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genetics
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Protein Precursors
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biosynthesis
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genetics
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Protein Renaturation
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Recombinant Fusion Proteins
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biosynthesis
;
genetics
;
isolation & purification
9.Small interference RNA targeting vascular endothelial growth factor gene effectively attenuates retinal neovascularization in mice model.
Yi-chun KONG ; Tianjin Eye INSTITUTE ; Bei SUN ; Kan-xing ZHAO ; Mei HAN ; Yu-chuan WANG
Chinese Medical Journal 2013;126(8):1440-1444
BACKGROUNDThe mechanism of retinal neovascularization is not understood completely. Many growth factors are involved in the process of retinal neovascularization, such as vascular endothelial growth factor (VEGF) and pigment epithelium-deprived factor (PEDF), which are the representatives of angiogenic and antiangiogenic molecules respectively. Oxygen induced retinopathy (OIR) is a useful model to investigate retinal neovascularization. The present study was conducted to investigate the feasibility of small interference RNA (siRNA) targeting VEGF gene in attenuating oxygen induced retinopathy (OIR) by regulating VEGF to PEDF ratio (VEGF/PEDF).
METHODSIn vitro, cultured EOMA cells were transfected with VEGF-siRNA (psi-HI(TM)/EGFP/VEGF siRNA) and Lipofectamine(TM) 2000 for 24, 48, and 72 hours, respectively. Expression of VEGF mRNA was evaluated by real time polymerase chain reaction (PCR) and the level of VEGF protein was analyzed by Western blotting. In vivo, OIR model mice were established, the mice (C57BL/6J) received an intra-vitreal injection of 1 µl of mixture of psi-HI(TM)/EGFP/VEGF siRNA and Lipofectamine 2000. Expressions of retinal VEGF and PEDF protein were measured by Western blotting, retinal neovascularization was observed by fluorescein angiography, and quantified.
RESULTSIn vitro psi-HI(TM)/EGFP/VEGF siRNA treatment significantly reduced VEGF mRNA and protein expression. In vivo, with decreased VEGF and VEGF-PEDF ratio, significant attenuation of neovascular tufts, avascular regions, tortuous, and dilated blood vessels were observed in the interfered animals.
CONCLUSIONSVEGF plays an important role in OIR, and the transfection of VEGF-siRNA can effectively downregulate VEGF expression in vivo, accompanied by the downregulation of VEGF-PEDF ratio, and simultaneous attenuation of retinal neovascularization was also observed. These findings suggest that VEGF/PEDF may serve as a potential target in the treatment of retinal neovascularization and RNA interference targeting VEGF expression, which represents a possible therapeutic strategy.
Animals ; Eye Proteins ; analysis ; Mice ; Mice, Inbred C57BL ; Nerve Growth Factors ; analysis ; RNA, Small Interfering ; genetics ; Retinal Neovascularization ; therapy ; Serpins ; analysis ; Vascular Endothelial Growth Factor A ; analysis ; genetics ; physiology
10.Advances in the studies of Kallmann syndrome.
National Journal of Andrology 2006;12(7):647-649
Kallmann syndrome (KS) is a rare hereditary disease. It is characterized by hypogonadotrophic hypogonadism in association with anosmia or hyposmia. At present, three modes of inheritance and genes related to KS have been identified. This review focuses on the clinical diagnosis and advances in the studies of the pathogenesis gene for Kallmann syndrome.
Diagnosis, Differential
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Extracellular Matrix Proteins
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genetics
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Humans
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Kallmann Syndrome
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diagnosis
;
genetics
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therapy
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Male
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Nerve Tissue Proteins
;
genetics
;
Rare Diseases
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Receptor, Fibroblast Growth Factor, Type 1
;
genetics