Universal relation between the conditional auto-correlation function and the cross-correlation function of biphotons.

We systematically studied the relation between the conditional auto-correlation function (CACF) and cross-correlation function (CCF) of biphotons or pairs of single photons. The biphotons were generated from a heated atomic vapor via the spontaneous four-wave mixing (SFWM) process. In practical usage, one single photon of a pair is utilized as the heralding photon, and another is employed as the heralded photon. Motivated by the data of CACF of the heralded photons versus CCF, we proposed a universal formula to predict the CACF. The derived formula was based on general theory and is also valid for the biphoton generation process of spontaneous parametric down-conversion (SPDC). With the formula, we utilized the experimentally determined parameters to predict CACFs, which can well agree with the measured CACFs. The proposed formula enables one to quantitatively know the CACF of heralded single photons without the measurement of Hanbury-Brown-Twiss-type three-fold coincidence count. This study provides a better understanding of biphoton generation using the SFWM or SPDC process. Our work demonstrates a valuable tool for analyzing a vital property of how the heralded photons are close to Fock-state single photons.

Gender-Specific Association of Handgrip Strength with Type 2 Diabetes Mellitus in Chinese Han Older Adults.

Purpose: We aimed to analyze the relationship between handgrip strength/relative handgrip strength among older Han adults with type 2 diabetes mellitus (T2DM) by gender to determine the optimal cut-off value of grip strength for older adults. Methods: A multi-stage sampling method was used to conduct a questionnaire survey and physical examination of 6128 older adults in Anhui Province. Chi-squares tests, t-tests, analysis of variance, and logistic regression analysis were used to analyze the association between handgrip strength/relative handgrip strength and T2DM between the sexes. The decision tree model (CRT) was used to explore the predictive value of handgrip strength /relative handgrip strength on T2DM. Results: There was an association between handgrip strength and T2DM (P = 0.006, OR = 0.985, 95% CI = 0.975, 0.996), which was found in females (P = 0.013, OR = 0.978, 95% CI = 0.961, 0.995) but not in males (P = 0.125, OR = 0.989, 95% CI = 0.976, 1.003). Relative handgrip strength was also correlated with T2DM (P = 0.014, OR = 0.730, 95% CI = 0.568, 0.939), which was found in females (P = 0.003, OR = 0.534, 95% CI = 0.352, 0.809) but not in males (P = 0.432, OR = 0.879, 95% CI = 0.638, 1.212). The incidence of T2DM in elderly females with hypertension who were uneducated and with a handgrip strength of <17.350 kg was 24.3% (115 cases), whereas that in elderly females with hypertension and a relative handgrip strength of <0.240 was 29.0% (127 cases). Conclusion: According to our results, handgrip strength and relative handgrip strength were associated with T2DM. People with hypertension had a higher risk of T2DM in women with a handgrip strength of ≤ 17.350kg and a relative grip strength of ≤ 0.240. Further research is needed to validate the effectiveness of this cut-off for implementing interventions and avoiding risks.

Generation of sub-MHz and spectrally-bright biphotons from hot atomic vapors with a phase mismatch-free scheme.

We utilized the all-copropagating scheme, which maintains the phase-match condition, in the spontaneous four-wave mixing (SFWM) process to generate biphotons from a hot atomic vapor. The linewidth and spectral brightness of our biphotons surpass those of the biphotons produced with the hot-atom SFWM in the previous works. Moreover, the generation rate of the sub-MHz biphoton source in this work can also compete with those of the sub-MHz biphoton sources of the cold-atom SFWM or cavity-assisted spontaneous parametric down conversion. Here, the biphoton linewidth is tunable for an order of magnitude. As we tuned the linewidth to 610 kHz, the generation rate per linewidth is 1,500 pairs/(s·MHz) and the maximum two-photon correlation function, gs,as(2), of the biphotons is 42. This gs,as(2) violates the Cauchy-Schwarz inequality for classical light by 440 folds, and demonstrates that the biphotons have a high purity. By increasing the pump power by 16 folds, we further enhanced the generation rate per linewidth to 2.3×104 pairs/(s·MHz), while the maximum gs,as(2) became 6.7. In addition, we are able to tune the linewidth down to 290±20 kHz. This is the narrowest linewidth to date among all single-mode biphoton sources of room-temperature and hot media.

Expression levels of the α7 nicotinic acetylcholine receptor in the brains of patients with Alzheimer's disease and their effect on synaptic proteins in SH-SY5Y cells.

Alzheimer's disease (AD) is a chronic neurodegenerative, and abnormal aggregation of the neurotoxic ß amyloid (Aß) peptide is an early event in AD. The present study aimed to determine the correlation between the nicotinic acetylcholine receptor α7 subunit (α7 nAChR) and Aß in the brains of patients with AD, and to investigate whether the increased expression levels of the α7 nAChR could alter the neurotoxicity of Aß. The expression levels of α7 nAChR and Aß in the brains of patients with AD and healthy brains were analyzed using immunofluorescence. Moreover, SH­SY5Y cells were used to stably overexpress or silence α7 nAChR expression levels, prior to the treatment with or without 1 µmol/l Aß1­42 oligomer (AßO). The mRNA and protein expression levels of α7 nAChR, synaptophysin (SYP), postsynaptic density of 95 kDa (PSD­95) and synaptosomal­associated protein of 25 kDa (SNAP­25) were subsequently analyzed using reverse transcription­quantitative PCR and western blotting. In addition, the concentration of acetylcholine (ACh) and the activity of acetylcholinesterase (AChE) were analyzed using spectrophotometry, while the cell apoptotic rate was determined using flow cytometry. The expression of Aß in the brains of patients with AD was found to be significantly increased, whereas the expression of α7 nAChR was significantly decreased compared with the healthy control group. In vitro, the expression levels of α7 nAChR were significantly increased or decreased following the overexpression or silencing of the gene, respectively. Consistent with these observations, the mRNA and protein expression levels of SYP, PSD­95 and SNAP­25 were also significantly increased following the overexpression of α7 nAChR and decreased following the genetic silencing of the receptor. In untransfected or negative control cells, the expression levels of these factors and the apoptotic rate were significantly reduced following the exposure to AßO, which was found to be attenuated by α7 nAChR overexpression, but potentiated by α7 nAChR RNA silencing. However, no significant differences were observed in either the ACh concentration or AChE activity following transfection. Collectively, these findings suggested that α7 nAChR may protect the brains of patients with AD against Aß, as α7 nAChR overexpression increased the expression levels of SYP, SNAP­25 and PSD­95, and attenuated the inhibitory effect of Aß on the expression of these synaptic proteins and cell apoptosis. Overall, this indicated that α7 nAChR may serve an important neuroprotective role in AD.

Effect of Gastrodiae Rhizoma Powder on Prevention of Dementia and Oxidation Resistance in Mice / 中国实验方剂学杂志

Objective::To study whether long-term administration of Gastrodiae Rhizoma powder can improve the learning and memory ability of APPswe/PSldE9 double transgenic (APP/PS1) Alzheimer' s disease(AD) model mice and delay the progress of AD whether these effects are related to the regulation of antioxidant stress pathway in Kelch-like epoxylopropylamine-related protein 1(Keap1)-nuclear factor E2 related factor 2 (Nrf2)/heme oxygenase(HO)-1, and further explore the neuroprotective mechanism of Gastrodiae Rhizoma powder and its role in the prevention and treatment of AD. Method::APP/PS1 double transgenic mice model, the mice consisted of five groups: normal, normal administration group, model group, Gastrodiae Rhizoma powder prevention group, Gastrodiae Rhizoma powder treatment group.The mice in the normal administration group and the Gastrodiae Rhizoma powder prevention group were given the same dose of Gastrodiae Rhizoma powder (1.5 g·kg-1) daily at the age of 8 weeks.The normal group and model group were given the same amount of normal saline at the same time, until 24 weeks old, Morris water maze was used to test the learning and memory ability of mice, and the treatment group was treated with Gastrodiae Rhizoma powder at 22 weeks old.The mice were given the same dose of Gastrodiae Rhizoma powder (1.5 g·kg-1) every day for 2 weeks.The number of crossing platform, escape latency and platform residence time of mice were detected by Morris water maze from 24 weeks old to 24 weeks old.RNA, Real-time PCR was extracted from mouse hippocampus to detect the mRNA level of Nrf2, HO-1, Keap1, and Western blot was used to detect the expression of Nrf2, HO-1, Keap1 protein in mouse hippocampus. Result::Compared with normal group, the water maze test showed that the learning and memory ability of model group was lower than that of the model group (P<0.01), and the learning and memory ability of Gastrodiae Rhizoma powder prevention group and Gastrodiae Rhizoma powder treatment group was significantly higher than that of model group (P<0.01). Compared with normal group, the levels of Nrf2, HO-1 and protein in the hippocampus in model group decreased in varying degrees (P<0.05). Compared with model group, Gastrodiae Rhizoma powder prevented Nrf2, in the hippocampus of mice in model group.The level of HO-1 in mRNA and protein increased in different degrees (P<0.05, P<0.01). Levels of Nrf2, HO-1 mRNA in Gastrodiae Rhizoma powder treatment group was significantly higher than that in Gastrodiae Rhizoma powder group (P<0.05). There was no significant difference in the expression of Nrf2, HO-1 protein.There was no significant difference in mRNA and protein levels of Keap1 among different groups. Conclusion::Morris water maze test and other results showed that Gastrodiae Rhizoma powder could improve the learning and memory ability of APP/PS1 mice, and it may enhance the expression of downstream antioxidant genes by regulating Keap1-Nrf2/HO-1 pathway.And then improve the learning and memory ability of APP/PS1 mice.

Clinical diagnosis and treatment for age-related macular degeneration: a brief analysis of the guidelines / 中国医师杂志

Age-related macular degeneration (AMD) is a common cause of blindness among people over 65 in developed countries.With the rapidity of population aging process,the prevalence of AMD will be further increased.The application of anti-vascular endothelial factor growth medicine in ophthalmology has made great progress in the therapeutic effect and prognosis of wet AMD.In this context,many countries and regions have successively formulated guidelines for the AMD clinical diagnosis and treatment,especially the United States,Europe and Australia.Through the analysis of AMD clinical guidelines of American Academy of Ophthalmology (AAO) in 2015,and by comparing it with AMD analysis and treatment guidelines of European Society of Retina Specialists (EURETINA) in 2014,this paper provides an accurate,effective and comprehensive diagnosis strategy and lays a foundation for providing AMD patients with quality diagnosis and treatment plans.

Effect of nuclear factor-κB signaling pathway on the expression of hypoxia-inducible factor-1 and vascular endothelial growth factor in nasal polyp cells / 新乡医学院学报

Objective To observe the effect of nuclear factor-κB (NF-κB) signaling pathway on the expression of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) in nasal polyp cells under hypoxic cultivation,and to investigate the relationship between NF-κB signaling pathway and the development of nasal polyp.Methods The nasal polyp and inferior turbinate tissue specimens were collected in the First Affiliated Hospital of Jiamusi University from January 2012 to December 2014.The nasal polyp and inferior turbinate tissues were taken to obtain nasal polyp cells and inferior turbinate cells,then the cells were cultured in primary culture,and the cells were cultured under hypoxia when they grew to 90％.When the cells were cultured in vitro to 90％,the NF-κB inhibitor BAY11-7082 was added (inhibitor intervention group),the other cells without inhibitor were used as controls (no inhibitor group),then the cells in the two groups were cultured under hypoxia.The cells were collected when they were cultured for 0,3,6 and 9 hours,respectively;and the expression of HIF-1α,VEGF and NF-κB p65 protein in the cells were detected by Western blot.Results Compared with 0 hour,the expression of HIF-1α,VEGF and NF-κB p65 protein in nasal polyp cells increased significantly after 3,6 and 9 hours of hypoxic cultivation (P ＜ 0.05);however,the expression of HIF-1α,VEGF and NF-κB p65 protein in inferior turbinate cells was not statistically significant (P ＞ 0.05).The expression of HIF-1α,VEGF and NF-κB p65 protein in nasal polyposis cells after 6 hours of hypoxic cultivation was significantly higher than that after 3 and 9 hours of hypoxic cultivation (P ＜ 0.05);but there was no significant difference in the expression of HIF-1α,VEGF and NF-κB p65 protein in nasal polyp cells between 3 and 9 hours of hypoxic cultivation (P ＞ 0.05).Compared with 0 hour,the expression of HIF-1α and VEGF protein in nasal polyp cells of no inhibitor group increased significantly after 3,6 and 9 hours of hypoxic cultivation (P ＜ 0.05);and the expression of HIF-1α and VEGF protein in nasal polyp cells after 6 hours of hypoxic cultivation was significantly higher than that after 3 and 9 hours of hypoxic cultivation in no inhibitor group (P ＜ 0.05).But there was no significant difference in the expression of HIF-1α and VEGF protein in nasal polyp cells of no inhibitor group between 3 and 9 hours of hypoxic cultivation (P ＞ 0.05).There was no significant difference in the expression of HIF-1 α and VEGF protein in nasal polyp cells of the inhibitor intervention group among 0,3,6 and 9 hours of hypoxic cultivation (P ＞ 0.05).There was no significant difference in the expression of HIF-1α and VEGF protein in nasal polyp cells between no inhibitor group and inhibitor intervention group at 0 hour of hypoxic cultivation (P ＞0.05).The expression of HIF-1α and VEGF protein in nasal polyp cells of inhibitor intervention group was significantly lower than that of no inhibitor group after 3,6 and 9 hours of hypoxic cultivation (P ＜ 0.05).Conclusion The expression of HIF-1α,VEGF and NF-κB p65 protein increased in nasal polyp cells under hypoxia condition.NF-κB signaling pathway may mediate hypoxia-induced HIF-1α and VEGF protein expression,and participate in the occurrence and development of nasal polyp.

Result analysis on testing reproductive tract infection pathogen of women / 中国现代医生

Objective To investigate the effect of differential diagnosing female genital tract infection diseases by using leucorrhea microscopy method, bacteria pre-enzyme spectrum analysis method and the joint method. Methods Both leucorrhea microscopy method and bacteria pre-enzyme spectrum analysis method were respectively used to detect the vaginal discharges of 4130 women from our hospital outpatient service, and test results combined with clinical manifes-tations and treatment effect were also analyzed and compared summarily. Results 1976 patients were founded by using the leucorrhea microscopy method (detection rate and accuracy rate was 47.8%,71.6% respectively); 1302 cases were founded by using bacteria pre-enzyme spectrum analysis method (detection rate and accuracy rate was 31.5%, 58.1%respectively);2622 cases were detected by using the joint detection method (detection rate and accuracy rate was 63.5%, 91.1% respectively). Conclusion The joint method has the advantages of both leucorrhea microscopy method and bac-teria pre-enzyme spectrum analysis method, and can accurately identify five common kinds of female reproductive tract infection diseases with one sample for once.

[Case-control study on the relationship of chronic low back pain of facet joint origin with the distribution of nerve endings and neuropeptide: a quantitative histological analysis].

OBJECTIVE: To investigate the density and distribution of nerve endings and neuropeptide Y (NPY) in lumbar facet joints of patients with low back pain. METHODS: Fifteen patients without low back pain were selected as control group (group A). Facet joint samples in group A were obtained during the operation or lumbar spinal canal tumor they suffered from. Those patients with low back pain were divided into three groups according to their different origins of pain, such as not from facet joint (group B, 15 patients) ,from facet joint only (group C, 20 patients), or from facet joint partially (group D, 20 patients). Different origins were determined by VAS after facet joint block. The density and distribution of nerve ending and neuropeptide in the capsular tissues were analyzed by a modified gold chloride staining and immunochemistry respectively. RESULTS: Compared with the ones in group A and B, the fact joints in group C and D were more inclined to be degenerated and got more nerve endings. NPY was expressed mainly in the facet joint of patients with low back pain in group C and D. In addition, there was a significant relationship between the distribution of nerve endings and NPY expression,while none of them were related with MRI Fujiwara grade of facet joint. CONCLUSION: These results suggest that the number of mechanoreceptors, neural sprouting and secreted peptides in the facet joint capsules vary with the change of mechanical or nociceptive stimulation, which may promote the development of low back pain in return.

Uniaxial cyclic stretch promotes osteogenic differentiation and synthesis of BMP2 in the C3H10T1/2 cells with BMP2 gene variant of rs2273073 (T/G).

Ossification of the posterior longitudinal ligament of the cervical spine (OPLL) is characterized by the replacement of ligament tissues with ectopic bone formation, and this result is strongly affected by genetic and local factors. Two single nucleotide polymorphisms (SNPs) of rs2273073 (T/G) and rs235768 (A/T) of bone morphogenetic protein 2 (BMP2) gene which are associated with OPLL have been reported in our previous report. In this study, we confirmed the connection in 18 case samples analysis of BMP2 gene in OPLL patients; additionally, it was also shown from the OPLL patients with ligament tissues that enchondral ossification and expression of BMP2 were significantly higher compared with the non-OPLL patients by histological examination, immunohistochemistry and Western blotting analysis. To investigate the underlying mechanism, we studied the effect of SNPs in cell model. The C3H10T1/2 cells with different BMP2 gene variants were constructed and then subjected to uniaxial cyclic stretch (0.5 Hz, 10% stretch). In the presence of mechanical stress, the expression of BMP2 protein in C3H10T1/2 cells transfected by BMP2 (rs2273073 (T/G)) and BMP2 (rs2273073 (T/G), rs235768 (A/T)) were significantly higher than the corresponding static groups (P<0.05). In conclusion, these results suggested that BMP2 gene variant of rs2273073 (T/G) could not only increase cell susceptibility to bone transformation similar to pre-OPLL change, but also increase the sensibility to mechanical stress which might play an important role during the progression of OPLL.

Case-control study on the relationship of chronic low back pain of facet joint origin with the distribution of nerve endings and neuropeptide: a quantitative histological analysis / 中国骨伤

<p><b>OBJECTIVE</b>To investigate the density and distribution of nerve endings and neuropeptide Y (NPY) in lumbar facet joints of patients with low back pain.</p><p><b>METHODS</b>Fifteen patients without low back pain were selected as control group (group A). Facet joint samples in group A were obtained during the operation or lumbar spinal canal tumor they suffered from. Those patients with low back pain were divided into three groups according to their different origins of pain, such as not from facet joint (group B, 15 patients) ,from facet joint only (group C, 20 patients), or from facet joint partially (group D, 20 patients). Different origins were determined by VAS after facet joint block. The density and distribution of nerve ending and neuropeptide in the capsular tissues were analyzed by a modified gold chloride staining and immunochemistry respectively.</p><p><b>RESULTS</b>Compared with the ones in group A and B, the fact joints in group C and D were more inclined to be degenerated and got more nerve endings. NPY was expressed mainly in the facet joint of patients with low back pain in group C and D. In addition, there was a significant relationship between the distribution of nerve endings and NPY expression,while none of them were related with MRI Fujiwara grade of facet joint.</p><p><b>CONCLUSION</b>These results suggest that the number of mechanoreceptors, neural sprouting and secreted peptides in the facet joint capsules vary with the change of mechanical or nociceptive stimulation, which may promote the development of low back pain in return.</p>

[Effects of α3 neuronal nicotinic acetylcholine receptor on cell apoptosis and p38 MAPK signal transduction pathway in SH-SY5Y cells].

OBJECTIVE: To investigate the effects of α3 neuronal nicotinic acetylcholine receptor (nAChR) on apoptosis and p38 signal transduction pathway in SH-SY5Y cells and to assess the roles of α3 nAChR in the pathogenesis of Alzheimer's disease (AD). METHODS: The levels of α3 nAChR mRNA and protein were measured by real-time PCR and Western blot, respectively, in SH-SY5Y cells transfected with α3 nAChR siRNA. The mRNA level of bcl-2 and bax was measured by the real-time PCR. The siRNA transfected SH-SY5Y cells and control were then treated with 10 µmol/L Aß25-35 for another 48 h, and the change in apoptotic rate and the levels of p-p38 and p38 were measured by flow cytometry and Western blot. Subsequently these SH-SY5Y cells were exposed to a blocker of p38 protein, and the apoptotic rate was measured again. RESULTS: Compared to the controls, the expression of α3 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with α3 nAChR siRNA decreased by 95% and 86%, respectively; the mRNA levels of bax increased 2.11 times and that for bcl-2 decreased 0.53 times. The apoptotic rate was unaffected (3.40% ± 0.20%); but it increased after Aß25-35 treatment (24.52% ± 1.59%); the level of p-p38 protein also increased by 178% in the α3 nAChR inhibited cells treated with Aß25-35. Compared to controls, the Aß25-35-treated SH-SY5Y cells and the Aß25-35-treated and siRNA-transfected cells both showed a reduction in apoptosis after treatment with p38 blocker, especially in the former. CONCLUSION: The siRNA silencing of α3 nAChR mRNA may enhance the effect of Aß25-35 on the cell apoptosis by increasing the levels of p38 protein and bax mRNA and decreasing the level of bcl-2 mRNA, which may play a role in the pathogenesis of AD.

Preventing expression of the nicotinic receptor subunit α7 in SH-SY5Y cells with interference RNA indicates that this receptor may protect against the neurotoxicity of Aß.

The present aim was to characterize the influence of the α7 nicotinic acetylcholine receptor (nAChR) on BACE, the enzyme that cleaves the amyloid precursor protein (APP) at the ß-site, as well as on the oxidative stress induced by amyloid-ß peptide (Aß). To this end, human neuroblastoma SH-SY5Y cells were transfected with siRNAs targeting the α7 nAChR subunit and/or exposed to Aß1-42. For α7 nAChR, BACE1 (cleaving at the ß-site of APP) and BACE2 (cleaving within the Aß domain), α-secretase (ADAM10), and the two components of γ-secretase, PS and NCT, the mRNA and protein levels were determined by real-time PCR and Western blotting, respectively. The level of Aß1-42 in the cell culture medium was determined by an ELISA procedure. The extent of lipid peroxidation and activities of superoxide dismutase (SOD) and glutathione peroxidase (GSH-Px) were assayed spectrophotometrically. In the transfected SH-SY5Y cells, expression of α7 nAChR was reduced; the level of BACE1 increased and that of BACE2 decreased; the amount of ADAM10 lowered; and the level of PS raised. Moreover, the level of Aß1-42 in the culture medium was elevated. Treatment of non-transfected cells with Aß elevated the level of malondialdehyde (MDA) and lowered the activities of SOD and GSH-Px and these changes were potentiated by inhibiting expression of α7 nAChR. These results indicate that α7 nAChR plays a significant role in amyloidogenic metabolism of APP and the oxidative stress evoked by Aß, suggesting that this receptor might help protect against the neurotoxicity of Aß.

Effects of α3 neuronal nicotinic acetylcholine receptor on cell apoptosis and p38 MAPK signal transduction pathway in SH-SY5Y cells / 中华病理学杂志

<p><b>OBJECTIVE</b>To investigate the effects of α3 neuronal nicotinic acetylcholine receptor (nAChR) on apoptosis and p38 signal transduction pathway in SH-SY5Y cells and to assess the roles of α3 nAChR in the pathogenesis of Alzheimer's disease (AD).</p><p><b>METHODS</b>The levels of α3 nAChR mRNA and protein were measured by real-time PCR and Western blot, respectively, in SH-SY5Y cells transfected with α3 nAChR siRNA. The mRNA level of bcl-2 and bax was measured by the real-time PCR. The siRNA transfected SH-SY5Y cells and control were then treated with 10 µmol/L Aβ25-35 for another 48 h, and the change in apoptotic rate and the levels of p-p38 and p38 were measured by flow cytometry and Western blot. Subsequently these SH-SY5Y cells were exposed to a blocker of p38 protein, and the apoptotic rate was measured again.</p><p><b>RESULTS</b>Compared to the controls, the expression of α3 nAChR at mRNA and protein levels in the SH-SY5Y cells transfected with α3 nAChR siRNA decreased by 95% and 86%, respectively; the mRNA levels of bax increased 2.11 times and that for bcl-2 decreased 0.53 times. The apoptotic rate was unaffected (3.40% ± 0.20%); but it increased after Aβ25-35 treatment (24.52% ± 1.59%); the level of p-p38 protein also increased by 178% in the α3 nAChR inhibited cells treated with Aβ25-35. Compared to controls, the Aβ25-35-treated SH-SY5Y cells and the Aβ25-35-treated and siRNA-transfected cells both showed a reduction in apoptosis after treatment with p38 blocker, especially in the former.</p><p><b>CONCLUSION</b>The siRNA silencing of α3 nAChR mRNA may enhance the effect of Aβ25-35 on the cell apoptosis by increasing the levels of p38 protein and bax mRNA and decreasing the level of bcl-2 mRNA, which may play a role in the pathogenesis of AD.</p>

LIPUS enhance elongation of neurites in rat cortical neurons through inhibition of GSK-3beta.

OBJECTIVE: Low-intensity pulsed ultrasound (LIPUS) has been reported to enhance proliferation and to alter protein production in various kinds of cells. In the present study, we measured the neurites length after LIPUS treatment to define the effectiveness of LIPUS stimulation on neurons, and then we examined the acticity of GSK-3beta to study the intracellular mechanism of neurite's outgrowth. METHODS: LIPUS was applied to cultured primary rat cortical neurons for 5 minutes every day with spatial- and temporal average intensities (SATA) of 10 mW/cm(2), a pulse width of 200 microseconds, a repetition rate of 1.5 KHz, and an operation frequency of 1 MHz. Neurons were photographed on the third day after LIPUS treatment and harvested at third, seventh, and tenth days for immunoblot and semi-quantitative RT-PCR analysis. RESULTS: Morphology change showed that neurite extension was enhanced by LIPUS. There was also a remarkable decrease of proteins, including p-Akt, p-GSK-3beta, and p-CRMP-2, observed on the seventh and tenth days, and of GSK-3beta mRNA expression, observed on the seventh day, in neurons treated with LIPUS. CONCLUSION: LIPUS can enhance elongation of neurites and it is possible through the decreased expression of GSK-3beta.

LIPUS enhance elongation of neurites in rat cortical neurons through inhibition of GSK-3beta / 生物医学与环境科学（英文）

<p><b>OBJECTIVE</b>Low-intensity pulsed ultrasound (LIPUS) has been reported to enhance proliferation and to alter protein production in various kinds of cells. In the present study, we measured the neurites length after LIPUS treatment to define the effectiveness of LIPUS stimulation on neurons, and then we examined the acticity of GSK-3beta to study the intracellular mechanism of neurite's outgrowth.</p><p><b>METHODS</b>LIPUS was applied to cultured primary rat cortical neurons for 5 minutes every day with spatial- and temporal average intensities (SATA) of 10 mW/cm(2), a pulse width of 200 microseconds, a repetition rate of 1.5 KHz, and an operation frequency of 1 MHz. Neurons were photographed on the third day after LIPUS treatment and harvested at third, seventh, and tenth days for immunoblot and semi-quantitative RT-PCR analysis.</p><p><b>RESULTS</b>Morphology change showed that neurite extension was enhanced by LIPUS. There was also a remarkable decrease of proteins, including p-Akt, p-GSK-3beta, and p-CRMP-2, observed on the seventh and tenth days, and of GSK-3beta mRNA expression, observed on the seventh day, in neurons treated with LIPUS.</p><p><b>CONCLUSION</b>LIPUS can enhance elongation of neurites and it is possible through the decreased expression of GSK-3beta.</p>

Effect of low-intensity pulsed ultrasound on the expression of neurotrophin-3 and brain-derived neurotrophic factor in cultured Schwann cells.

It is generally known that low-intensity pulsed ultrasound (LIPUS) accelerates peripheral nerve tissue regeneration. However, the precise cellular mechanism involved is still unclear. The purpose of this study was to determine how the Schwann cells respond directly to LIPUS stimuli. Thus, we investigated the effect of LIPUS on cell proliferation, neurotrophin-3 (NT-3), and brain-derived neurotrophic factor (BDNF) mRNA expression in rat Schwann cells. Schwann cells were enzymatically isolated from postnatal 1-3 day rat sciatic nerve tissue and cultured in the six-well plate. The ultrasound was applied at a frequency of 1 MHz and an intensity of 100 mW/cm(2) spatial average temporal average for 5 minutes/day. The control group was cultured in the same way but without the administration of ultrasound. Immunohistochemistry demonstrated that more than 98% of the experimental and control cells were positive for S-100, NT-3, and BDNF. With 5-bromo-2'-deoxyuridine (BrdU) assay, the stimulated cells also exhibited an increase in the rate of cell proliferation on days 4, 7, 10, and 14. Further investigation found that mRNA expression of NT-3 was significantly upregulated in experimental groups compared with the control 14 days after the LIPUS stimulation (the ratio of NT-3/beta-actin was 0.56 +/- 0.13 vs. 0.41 +/- 0.09, P < 0.01), whereas the mRNA expression of BDNF was significantly downregulated in experimental groups compared with the control (the ratio of BDNF/beta-actin was 0.51 +/- 0.05 vs. 0.60 +/- 0.08, P < 0.05). These results demonstrated that the application of LIPUS promotes cell proliferation and NT-3 gene expression in Schwann cells, and involved in the alteration of BDNF gene expression.