Objective To analyze the gene expression profiles in relation to the migration ability of adult human bone marrow-derived neural stem cells (Md-NSCs), and identify the genetic basis of the high migration potential of Md-NSCs in the central nervous system (CNS). Methods Adult human bone marrow stromal celIs(BMSCs) obtained from adult healthy volunteers were induced to differentiate into Md-NSCs in vitro, and the expressions of the genes related to cell invasiveness and metastasis were investigated by microarray analysis. Quantitative real-time PCR (RT-PCR) was used to verify the microarray results. Results The results of microarray analysis revealed significant overexpressions of the genes MMP1, MMP2, MMP17, ITGA3, RhoB, RhoC and RhoD in the Md-NSCs as compared with the expressions in fresh normal human adult bone marrow cells depleted of red blood cells. Quantitative RT-PCR verified the overexpression ofMMP2 gene by 2.84×100 folds, ITGA3 gene by 2.22×102folds, and RhoC gene by 4.92×100 folds. Conclusion The high migration potential of the Md-NSCs in the CNS is probably associated with the overexpression of the genes that promote cell invasiveness and metastasis. These overexpressed genes are also important oncogenes, and therefore the tumorigenicity of the Md-NSCs warrants further investigation.

OBJECTIVETo establish the triple-transgenic mouse model and study their biological characteristics by molecular biology, behavior and pathology.

METHODSHybrid the Tau and amyloid precursor protein (APP)/presenilins (PS1) transgenic mouse, the genotype of offspring mice were identified by PCR. Transcribed target genes were detected by RT-PCR. The protein expression of exogenous genes was detected by Western-blot. The pathological change of neurofibrillary tangles and senile plaque were observed by Bielschowsky silver staining and ABC immunohistochemical method. The changes time of learning and memory were observed by Morris water maze.

RESULTSAPP, PS1 and Tau genes were transcript in Tau/APP/PS1 mice. In 6 to 8 months old Tau/APP/PS1 mice, the neurofibrillary tangles and senile plaque could be found in cortex and hippocampus. In 6 months old Tau/APP/PS1 mice, the learning and memory abilities were worse.

CONCLUSIONWith the behavior change and pathological changes in Tau and beta-amyloid protein (AP), the Tau/APP/PS1 triple-transgenic mice can be used as a further study animal model of AD's pathogenesis and the target of drug treatment.

Alzheimer Disease ; pathology ; Amyloid beta-Protein Precursor ; genetics ; Animals ; Brain ; pathology ; Disease Models, Animal ; Learning ; Male ; Memory ; Mice ; Mice, Transgenic ; Neurofibrillary Tangles ; pathology ; Plaque, Amyloid ; pathology ; Presenilin-1 ; genetics ; tau Proteins ; genetics

OBJECTIVETo evaluate causes and clinical management of postoperative pain after total knee arthroplasty (TKA).

METHODSFrom January 2004 to June 2009, 41 patients (44 knees) with knee pain post TKA were treated. There were 9 male and 32 female patients aging from 51 to 84 years with a mean of 63.5 years. The diagnosis followed to Brown diagnostic system. One case of extraarticular pain was complex regional pain syndrome type 1 (CRPS-1) and underwent conservative treatment, the remaining 5 cases were treated by surgery. Three cases of joint instability, 1 case of patellar baja, 2 cases of soft tissue impingement caused by overhang of the prosthesis, 1 case of popliteal tendon impact underwent conservative treatment, the other 27 cases underwent surgical intervention. The patients were followed up and the Knee Society Score (KSS) knee score, pain visual analog scale (VAS) score were recorded.

RESULTSForty-one cases were followed up for 1 to 6 years. At the last time of follow-up, the 5 cases received surgical treatment to extra-articular pain showed VAS score as 2.5 ± 0.2, KSS clinical and functional score as 92.8 ± 2.6 and 89.0 ± 3.4. There was significantly difference compared with preoperative (P < 0.05). One case of CRPS-1 performed conservative treatment, the therapy was effective. In the infected 12 cases of intra-articular pain, 1 case received amputation, 3 cases received antibiotic bone cement insert, 8 cases received two stage revision. All infections were cured, and VAS score was 3.8 ± 0.2, KSS clinical score was 88.3 ± 4.6, function score was 85.0 ± 4.6 postoperatively, with significantly difference compared with preoperative (P < 0.05). In the 8 cases received conservative treatment in non-infected group, at the last time of follow-up, VAS score was 4.5 ± 0.4, KSS clinical and functional score was 85.4 ± 4.2 and 84.2 ± 2.3, with significantly difference compared with preoperative (P < 0.05). Fifteen cases underwent surgical treatment, at the last time of follow-up, VAS score was 3.4 ± 0.1, KSS clinical and functional score was 86.6 ± 5.4 and 87.1 ± 2.4, with significantly difference compared with preoperative (P < 0.05).

CONCLUSIONSPatients with knee pain post TKA need systematic assessment to identify the causes. Appropriate treatment due to the positive diagnosis generally lead to satisfactory results, surgical intervention with indefinite causes is strictly prohibited.

Aged ; Aged, 80 and over ; Arthroplasty, Replacement, Knee ; adverse effects ; Female ; Follow-Up Studies ; Humans ; Male ; Middle Aged ; Pain, Postoperative ; etiology ; therapy ; Retrospective Studies

ObjectiveThe objective of the present study was to compare the pregnancy and perinatal outcomes in women of advanced age between autologous fresh ET and FET cycles. MethodsA retrospective cohort study was conducted， which included 1 622 transfer cycles of 1 622 patients with ≥35 years of age. Patients who received their first embryo transfer cycle （IVF/ICSI treatment） between October 2015 and May 2020 in the center of reproductive medicine of the Third Affiliated Hospital of Sun Yat-sen University were included. Among them， 903 patients had 903 fresh ET cycles， and 719 freeze-all patients received 719 FET cycles. The baseline characteristics， pregnancy outcomes， complication rates， and perinatal outcomes were compared between the two groups. Logistic regression was performed to adjusted for confounding factors. ResultsWomen with advanced age and fresh embryo transfer had significantly higher clinical pregnancy rate ［40.2% vs. 36.6%，OR 95%CI：1.66 （1.323，2.078），P＜0.001］， live birth rate ［29.7% vs. 23.8%，OR 95%CI：1.93 （1.505，2.484），P＜0.001］， and multiple pregnancy rate ［25.1% vs. 12.2%，OR 95%CI：1.89 （1.114，3.190），P＝0.018］ than those with FET cycles. Fresh embryo transfer had significantly decreased miscarriage rate ［25.1% vs. 33.5%，OR 95%CI：0.53 （0.356，0.794），P＝0.002］， Caesarean delivery rate ［70.5% vs. 76.6%，OR 95%CI：0.60 （0.362，0.987），P＝0.044］， and hypertensive disorders of pregnancy ［2.2% vs. 4.6%，OR 95%CI：0.35 （0.128，0.947），P＝0.039］ than those of FET group. There were no significant differences in the rate of ectopic pregnancy， preterm delivery rate， gestational diabetes mellitus. and preterm premature rupture between the two groups （P＞0.05）. FET group had significantly higher mean new born birth weight ［（3 133 ± 612） g vs. （2 977 ± 609） g，P=0.006］. Further age-stratified analysis showed that the clinical pregnancy rate of fresh transfer was significantly higher than that of frozen embryo transfer in patients aged 35-37 years ［47.8% vs. 45.4%，OR 95%CI：1.68 （1.176，2.391），P =0.004］. The live birth rate of fresh embryo transfer in patients aged 35-37 years and 38-39 years was higher than that of frozen embryo transfer ［39.3% vs. 32.9%，OR 95%CI： 1.79 （1.239，2.581），P =0.002］， ［35.5% vs. 30.4%，OR 95%CI： 1.70 （1.023，2.809），P =0.040］ . ConclusionsPatients with advanced age obtain better pregnancy and neonatal outcomes by fresh embryo transfer than by frozen embryo transfer. Patients without contraindications should preferentially choose fresh embryo transfer to improve their pregnancy outcome， reduce the waiting time for embryo transfer， and to avoid the adverse outcomes which increase over time with advanced age.

BACKGROUNDPancreatic beta-cell apoptosis induced by lipotoxicity, to a large extent, contributes to the progression of type 2 diabetes. To investigate the mechanism of free fatty acid induced apoptosis, we aimed to study the effects of palmitic acid (PA) on the apoptosis and peroxisome proliferator-activated receptor γ coactivator-1α (PGC-1α) expression in βTC3 cells as well as the possible role of nuclear factor-κB (NF-κB) in this process.

METHODSHoechst 33258 was used to detect βTC3 cell apoptosis, which was induced by PA stimulation for 12 hours. PGC-1α expression was analyzed by reverse transcription polymerase chain reaction, IκB kinase β (IKKβ), IκBα, NF-κB-inducing kinase (NIK) and Rel-B expressions were analyzed by Western blotting. MG132 was employed to block the endogenous IκBα degradation before PA administration, and then its effect on PA-inducing cell apoptosis and PGC-1α mRNA expression was analyzed.

RESULTSSignificant increased cell apoptosis was found at the concentration of 0.5 mmol/L and 1.0 mmol/L PA administration. PA (0.5 mmol/L) could extensively reduced the expression of PGC-1α mRNA. After exposing βTC3 cells to 0.5 mmol/L PA for different time periods (0, 4, 6, 8, 10 and 12 hours), IKKβ protein expression increased while IκBα, NIK and Rel-B protein expression declined in a time-dependent manner. Pretreatment with MG132 to inhibit the degradation of IκBα, partially prevented the down-regulation of PGC-1α mRNA expression after 12-hour PA treatment in accordance with the decrease of PA induced apoptosis.

CONCLUSIONSNF-κB canonical pathway was activated in PA-mediated βTC3 cell apoptosis, whereas non-canonical pathway was inhibited. Reduced PGC-1α expression by PA in βTC3 cells could involve the activation of canonical NF-κB pathway, so as to deteriorate the PA induced apoptosis.

Apoptosis ; drug effects ; Cell Line ; Heat-Shock Proteins ; genetics ; metabolism ; Humans ; Insulin-Secreting Cells ; drug effects ; metabolism ; Leupeptins ; pharmacology ; NF-kappa B ; genetics ; metabolism ; Palmitic Acid ; pharmacology ; Peroxisome Proliferator-Activated Receptor Gamma Coactivator 1-alpha ; Reverse Transcriptase Polymerase Chain Reaction ; Signal Transduction ; drug effects ; Transcription Factors ; genetics ; metabolism

Nicotine enhances the function of learning and memory, but the underlying mechanism still remains unclear. Hippocampal long-term potentiation (LTP) is assumed to be a cellular mechanism of learning and memory. Our previous experiments showed that with the single pulses evoking 80% of the maximal population spike (PS) amplitude, nicotine (10 μmol/L) induced LTP-like response in the hippocampal CA1 region. In the present study, the nicotinic acetylcholine receptor (nAChR) subtypes and relevant neurotransmitter releases involved in LTP-like response induced by nicotine were investigated by extracellularly recording the PS in the pyramidal cell layer in the hippocampal CA1 region in vitro. LTP-like response induced by nicotine was blocked by mecamylamine (1 μmol/L) or κ-bungarotoxin (0.1 μmol/L), but not by dihydro-β-erythtroidine (DHβE, 10 μmol/L). Moreover, it was inhibited by propranolol (10 μmol/L), but not by phentolamine (10 μmol/L) or atropine (10 μmol/L). The results suggest that noradrenaline release secondary to the activation of κ-bungarotoxin-sensitive nAChRs participates in LTP-like response induced by nicotine in the hippocampal CA1 region.
Animals ; Bungarotoxins ; CA1 Region, Hippocampal ; physiology ; Long-Term Potentiation ; drug effects ; Nicotine ; pharmacology ; Norepinephrine ; secretion ; Receptors, Nicotinic ; metabolism

OBJECTIVETo investigate the toxicity of cationic liposome Lipofectamine 2000 (Lipo) in human pancreatic cancer Capan-2 cells.

METHODSCapan-2 cells were cultured in the presence of Lipo at toxic concentrations, and the cell growth, apoptosis and cell cycle changes were evaluated by cell counting and flow cytometry.

RESULTSThe concentrations of both Lipo and siRNA affected the transfection efficiency. In a transfection volume of 2 ml, the presence of 5 microl Lipo resulted in slowed growth of Capan-2 cells, which was especially obvious after 3 days (P<0.001). Prolonged culture of the transfected cells caused significant increases in early apoptotic cells (P<0.05) and in the damaged or necrotic cells (P<0.001), and resulted in reduced viable cells (P<0.01); these changes became obvious after a 48-hour culture, which also increased the ratio of G(0)/G(1) phase cells (P<0.05) and decreased those of G(2)/M phase cells (P<0.01), S phase cells (P<0.01), and the late apoptotic cells (P<0.05).

CONCLUSIONToxic concentrations of Lipo can affect the growth, apoptosis and cell cycles of Capan-2 cells in vitro, and this urges careful concentration selection when using Lipo for gene transfer into different cells.

Apoptosis ; drug effects ; Cations ; toxicity ; Cell Cycle ; drug effects ; Cell Line, Tumor ; Humans ; Lipids ; genetics ; toxicity ; Liposomes ; toxicity ; Pancreatic Neoplasms ; pathology ; RNA, Small Interfering ; genetics ; Transfection

OBJECTIVETo explore the anti-tumor efficacy of anti- vascular endothelial growth factor (VEGF) McAb 5-fluorouracil (5-FU) loaded polylactic acid (PLA) nanoparticles (NPS) in human gastric carcinoma xenografts of nude mice.

METHODSAnti-VEGF McAb 5-FU loaded PLA NPS were made by ultrasound emulsification. Nude mice model of human gastric carcinoma xenografts was established. Therapeutic effects of drugs on human gastric carcinoma xenografts and side effects concerned were observed.

RESULTSThe tumor inhibition rates of control group, nanosphere without 5-FU group, 5-FU (20 mg/kg) group, anti-VEGF McAb nanosphere without 5-FU group, anti-VEGF McAb group, nanosphere with 5-FU group, 5-FU (20 mg/kg) combined with anti-VEGF McAb group, anti-VEGF McAb 5-FU loaded nanosphere group was 0, 6.61%, 24.26%, 27.94%, 35.29%, 37.50%, 39.71% and 52.21% respectively, and there were no significant differences between anti-VEGF McAb 5-FU loaded nanosphere group and nanosphere group without 5-FU in WBC count, serum alanine transferase level or creatinine level. Compared with control group and anti-VEGF McAb 5-FU loaded nanosphere group, the 5-FU group decreased by 34.43% and 37.38% respectively in WBC count (P< 0.05), and increased by 93.17% and 66.56% respectively in alanine transferase. There were significant differences between experimental groups and control group in apoptosis index, especially between anti-VEGF McAb 5-FU loaded nanosphere group and control group (P< 0.05). The microvessel density (MVD) of experimental groups containing anti-VEGF McAb was significantly lower than that of control group or groups containing 5-FU (P< 0.05).

CONCLUSIONAnti-VEGF McAb 5-FU loaded nanosphere can increase the tumor inhibitory rate of 5-FU, induce apoptosis by inhibiting tumor angiogenesis with less side effect, and then enhance therapeutic effect, which indicate its potential as a novel, safe nano-tumor-targeting drug.

Animals ; Antibodies, Monoclonal ; administration & dosage ; pharmacology ; Antimetabolites, Antineoplastic ; administration & dosage ; pharmacology ; Cell Line, Tumor ; Drug Carriers ; Fluorouracil ; administration & dosage ; pharmacology ; Humans ; Lactic Acid ; administration & dosage ; pharmacology ; Mice ; Mice, Nude ; Nanoparticles ; Neovascularization, Pathologic ; Polyesters ; Polymers ; administration & dosage ; pharmacology ; Stomach Neoplasms ; drug therapy ; pathology ; Vascular Endothelial Growth Factor A ; administration & dosage ; pharmacology ; Xenograft Model Antitumor Assays

OBJECTIVETo establish homozygous transgenic mouse strain expressing human tau isoform with P301L mutation.

METHODSFive transgenic mice expressing human tau isoform with P301L mutation were obtained by microinjection into male nuclei. Homozygote and hemizygote were identified by PCR and real-time fluorescent quantitative PCR.

RESULTSNinety five homozygous transgenic mice were selected, and the results indicated that homozygous transgenic mice were superior to hemizygote in simulating the changes of biological characteristics.

CONCLUSIONExogenous gene tau is able to stably transmit to next generation and the combination of SYBR Green real-time fluorescent quantitative PCR with the traditional mating is a fast, reliable and economical way to screen homozygous and hemizygous transgenic mice.

Animals ; Female ; Homozygote ; Humans ; Male ; Mice ; Mice, Inbred C57BL ; Mice, Transgenic ; Microinjections ; Mutation ; tau Proteins ; genetics

OBJECTIVETo evaluate the application value of magnetic resonance diffusion-weighted imaging (DWI) combined with routine T2WI sequence in the determination of pathological complete response (pCR) after neoadjuvant therapy for rectal cancer.

METHODSClinical data of 51 cases with locally advanced mid-low rectal cancer undergoing neoadjuvant therapy plus radical resection in the Rectal Cancer Center at The Sixth Affiliated Hospital of Sun Yat-sen University from June 2012 to April 2013 were analyzed retrospectively. Magnetic resonance DWI and T2WI sequences scanning were performed within 1 week before neoadjuvant therapy and within 1 week before operation. Routine single T2WI sequence and DWI combined with T2WI sequence were used separately to predict the residual tumor and to compare with postoperative pathological examination. The prediction values of two methods were compared.

RESULTSOf 51 patients, 12 cases had pathological complete response (pCR). Prediction of DWI combined T2WI sequence was correct in 8 cases of pCR, whose sensitivity and specificity were higher than those of routine single T2WI sequence (66.7%, 94.9% vs. 33.3%, 84.6%). Prediction value of DWI combined T2WI sequence for pCR was significantly higher as compared to routine single T2WI sequence (AUC, 0.808 vs. 0.590, P=0.001).

CONCLUSIONCompared with the routine single T2WI sequence, DWI combined with T2WI sequence can improve the prediction accuracy of pathological complete response.

Adult ; Aged ; Aged, 80 and over ; Diffusion Magnetic Resonance Imaging ; Female ; Humans ; Male ; Middle Aged ; Neoadjuvant Therapy ; Predictive Value of Tests ; Rectal Neoplasms ; pathology ; therapy ; Retrospective Studies ; Sensitivity and Specificity