Enhancing precision in lung tumor ablation through innovations in CT-guided technique and angle control.

BACKGROUND: This retrospective study aimed to evaluate the precision and safety outcomes of image-guided lung percutaneous thermal ablation (LPTA) methods, focusing on radiofrequency ablation (RFA) and microwave ablation (MWA). The study utilized an innovative angle reference guide to facilitate these techniques in the treatment of lung tumors. METHODS: This study included individuals undergoing LPTA with the assistance of laser angle guide assembly (LAGA) at our hospital between April 2011 and March 2021. We analyzed patient demographics, tumor characteristics, procedure details, and complications. Logistic regressions were employed to assess risk factors associated with complications. RESULTS: A total of 202 patients underwent ablation for 375 lung tumors across 275 sessions involving 495 ablations. Most procedures used RFA, especially in the right upper lobe, and the majority of ablations were performed in the prone position (49.7%). Target lesions were at a median depth of 39.3 mm from the pleura surface, and remarkably, 91.9% required only a single puncture. Complications occurred in 31.0% of ablations, with pneumothorax being the most prevalent (18.3%), followed by pain (12.5%), sweating (6.5%), fever (5.0%), cough (4.8%), hemothorax (1.6%), hemoptysis (1.2%), pleural effusion (2.0%), skin burn (0.6%), and air emboli (0.2%). The median procedure time was 21 min. Notably, smoking/chronic obstructive pulmonary disease emerged as a significant risk factor for complications. CONCLUSION: The LAGA-assisted LPTA enhanced safety by improving accuracy and reducing risks. Overall, this investigation contributes to the ongoing efforts to refine and improve the clinical application of these thermal ablation techniques in the treatment of lung tumors.

Giant Interpectoral Lipoma Causing Venous Thoracic Outlet Syndrome: A Rare Case Presentation.

Lipomas are the most prevalent type of benign soft tissue tumors, primarily composed of adipocytes, and typically remain asymptomatic unless they reach a significant size. Although giant lipomas are infrequent, their occurrence on the chest wall, particularly in the interpectoral region, is exceedingly rare. We present a unique case of a 48-year-old man with a massive interpectoral lipoma measuring 19.4 × 12.9 × 9.4 cm, which resulted in venous thoracic outlet syndrome by compressing the subclavian vein. This case highlights the clinical challenges in diagnosing deep-seated chest wall lipomas and underscores the necessity of considering thoracic outlet syndrome as a potential complication, even in the absence of direct neural or arterial compression. The presentation of thoracic outlet syndrome can vary, and a comprehensive evaluation is imperative for accurate diagnosis and management.

Pretreatment with Lovastatin Improves Depression-Like Behavior After Traumatic Brain Injury Through Activation of the AMPK Pathway.

BACKGROUND: The beneficial effect of pretreatment with statins on traumatic brain injury (TBI)-induced depression and anxiety and its mechanism of action remain unclear. In this study, we combined epidemiological and experimental animal data to clarify this issue. METHODS: We used the Taiwan National Health Insurance database to identify patients who were diagnosed with TBI from 2000 to 2013 and compared patients with and without statin treatment matched by age, sex, and underlying comorbidities in a 1:1 ratio. The risk of developing depression and/or anxiety was compared between patients with and without a statin using Cox proportional hazards regression. We also used a rat model to assess the effect of lovastatin pretreatment on neurobehavioral and neuropathological changes following TBI. RESULTS: The risk of developing depression was lower in the 41,803 patients in the statin cohort than nonstatin cohort (adjusted hazard ratio, 0.91 [95% confidence interval, 0.83-0.99]). In animal models, the lovastatin group had significantly reduced infarct volume, decreased immobility time and latency to eat, a reduced number of Fluoro- Jade-positive cells and levels of glial fibrillary acidic protein and tumor necrosis factor-alpha, and increased adenosine monophosphate -activated protein kinase (AMPK) and its upstream kinase liver kinase B1 in the hippocampal dentate gyrus. These effects were blocked in AMPK inhibitor-pretreated TBI rats. CONCLUSIONS: Our epidemiological data showed that a decreased risk of depression was associated with statin pretreatment, which was supported by an animal study. The underlying mechanism for this appears to involve AMPK activation in the statin pretreatment-induced alleviation of TBI.

Velopharyngeal function change after two-jaw orthognathic surgery in patients with cleft: A study on 162 consecutive cases.

BACKGROUND: Orthognathic surgery (OGS) is a common intervention used to correct midfacial hypoplasia in patients with cleft. Previous studies have reported that LeFort I maxillary advancement may impact velopharyngeal function, but similar investigations focusing on two-jaw OGS have not been conducted. METHODS: A total of 162 consecutive patients with cleft lip and palate who underwent two-jaw OGS between 2015 and 2020 were enrolled. Clinical data were collected, and preoperative and postoperative skeletal measurements were obtained from cephalometric images. Velopharyngeal function was evaluated using perceptual analysis and nasopharyngoscopy. A logistic regression model was employed for the risk factors associated with changes in velopharyngeal function. RESULTS: After two-jaw OGS, 82.1% of patients showed no change in velopharyngeal function, while 3.7% experienced improvement and 14.2% exhibited worsening of function. In addition, the changes in velopharyngeal function were statistically significant comparing to the pre-OGS velopharyngeal status. A multivariable logistic regression revealed that the amount of maxillary advancement independently predicted the deterioration of post-OGS velopharyngeal function (odds ratio = 1.74, 95% confidence interval (CI) = 1.20-2.52, p = 0.004). The receiver operating characteristic curve based on maxillary advancement demonstrated good discrimination, with an area under the curve of 0.727 (95% CI = 0.62-0.83, p = 0.001). The Youden index was 4.27 mm. CONCLUSION: Despite the risk of velopharyngeal function deterioration in patients with cleft palate undergoing OGS, some individuals have experienced improved function following two-jaw OGS. The extent of maxillary advancement has a negative impact on the velopharyngeal function.

A Simple Scoring Model for Evaluation of Concomitant Vascular Injuries in Patients with Lower Extremity Open Fractures.

BACKGROUND: Timely diagnosis and management of concomitant vascular injuries is usually needed for the management of lower extremity open fractures. In the current study, a prediction model and simplified scoring system of vascular injuries were developed for the primary evaluation of patients with lower extremity open fractures. METHODS: Patients with lower extremity open fractures were retrospectively reviewed from 2017 to 2020. Multivariate logistic regression analysis was used to evaluate independent risk factors for concomitant vascular injuries in these patients using data collected from 2017 through 2019 and a prediction scoring model was created accordingly. Model performance was validated with data from 2020. RESULTS: In total, 949 patients with lower extremity open fractures (development cohort, 705 patients, 2017 through 2019; validation cohort, 244 patients, 2020) were enrolled. Concomitant vascular injuries occurred in 44 patients in the development cohort (6.2%). Three clinical variables were identified for a prediction scoring model with weighted points, including hard or soft vascular signs (3 points), segmental fractures (2 points), and degloving soft-tissue injury (1 point). The model showed good discrimination (area under the receiver operating characteristic curve, 0.928), calibration (Hosmer-Lemeshow test, P = 0.661), and precision (Brier score, 0.041). Subsequent management regarding different aspects (observation only, further imaging study, or direct surgical exploration) can thus be decided. The model also demonstrated good discrimination (area under the receiver operating characteristic curve, 0.949), good calibration (Hosmer-Lemeshow test, P = 0.174), and good precision (Brier score, 0.042) in the validation cohort. CONCLUSION: This model may guide the subsequent management of vascular injuries associated with lower extremity open fractures. CLINICAL QUESTION/LEVEL OF EVIDENCE: Risk, III.

Modulation of methamphetamine memory reconsolidation by neural projection from basolateral amygdala to nucleus accumbens.

Drug-associated conditioned cues promote subjects to recall drug reward memory, resulting in drug-seeking and reinstatement. A consolidated memory becomes unstable after recall, such that the amnestic agent can disrupt the memory during the reconsolidation stage, which implicates a potential therapeutic strategy for weakening maladaptive memories. The basolateral amygdala (BLA) involves the association of conditioned cues with reward and aversive valences and projects the information to the nucleus accumbens (NAc) that mediates reward-seeking. However, whether the BLA-NAc projection plays a role in drug-associated memory reactivation and reconsolidation is unknown. We used methamphetamine (MeAM) conditioned place preference (CPP) to investigate the role of BLA-NAc neural projection in the memory reconsolidation. Two weeks before CPP training, we infused adeno-associated virus (AAV) carrying the designer receptor exclusively activated by designer drugs (DREADD) or control constructs. We infused clozapine-N-oxide (CNO) after the recall test to manipulate the neural activity of BLA-NAc projections in mice. We found that after recall, DREADD-mediated inhibition of BLA neurons projecting to the NAc core blunted consolidated MeAM-associated memory. Inhibition of BLA glutamatergic nerve terminals in the NAc core 1 h after recall disrupted consolidated MeAM-associated memory. However, inhibiting this pathway after the time window of reconsolidation failed to affect memory. Furthermore, under the condition without memory retrieval, DREADD-mediated activation of BLA-NAc core projection was required for amnesic agents to disrupt consolidated MeAM-associated memory. Our findings provide evidence that the BLA-NAc pathway activity is involved in the post-retrieval processing of MeAM-associated memory in CPP.

Direct-to-implant breast reconstruction following nipple-sparing mastectomy: predictive factors of adverse surgical outcomes in Asian patients.

BACKGROUND: Direct-to-implant (DTI) breast reconstruction after nipple-sparing mastectomy (NSM) with the use of acellular dermal matrix (ADM) provides reliable outcomes; however, the use of ADM is associated with a higher risk of complications. We analyzed our experiences of post-NSM DTI without ADM and identified the predictive factors of adverse surgical outcomes. METHODS: Patients who underwent NSM and immediate DTI or two-stage tissue expander (TE) breast reconstruction from 2009 to 2020 were enrolled. Predictors of adverse endpoints were analyzed. RESULTS: There were 100 DTI and 29 TE reconstructions. The TE group had a higher rate of postmastectomy radiotherapy (31% vs. 11%; P=0.009), larger specimens (317.37±176.42 g vs. 272.08±126.33 g; P=0.047), larger implants (360.84±85.19 g vs. 298.83±81.13 g; P=0.004) and a higher implant/TE exposure ratio (10.3% vs. 1%; P=0.035). In DTI reconstruction, age over 50 years (odds ratio [OR], 5.43; 95% confidence interval [CI], 1.50-19.74; P=0.010) and a larger mastectomy weight (OR, 1.65; 95% CI, 1.08-2.51; P=0.021) were associated with a higher risk of acute complications. Intraoperative radiotherapy for the nipple-areolar complex increased the risk of acute complications (OR, 4.05; 95% CI, 1.07-15.27; P=0.039) and the likelihood of revision surgery (OR, 5.57; 95% CI, 1.25-24.93; P=0.025). CONCLUSIONS: Immediate DTI breast reconstruction following NSM is feasible in Asian patients with smaller breasts.

Factors Impacting Survival in Patients With Major Burn-Induced Acute Kidney Injury Postrenal Replacement Therapy: A Nationwide Study With 15 Years Follow-Up in Taiwan.

BACKGROUND: Major burn-induced acute kidney injury (AKI) causes extremely high mortality, even though renal replacement therapy (RRT) was thought of as the most efficient treatment. There was scanty research for investigating the characteristic of burn-AKI-RRT patients during intensive care. This study aims to investigate the factors impacting the survival outcomes in those burn-AKI-RRT cases. METHODS: Using the Taiwan National Health Insurance Research Database and its affiliated database, the Registry for Catastrophic Illness Patients, we defined a cohort composed of 171 patients encountering major burn-induced AKI and receiving RRT during burn care for a 15-year observation period. Demographic characteristic, comorbidities, total body surface area (TBSA), major procedures, and complications were analyzed to explore the factors affecting the survival outcomes during acute burn care and 1 year after discharge. RESULTS: Patients who underwent tracheostomy and skin grafting had higher survival rates during acute burn care (tracheostomy: mortality vs survival, 15.7% vs 30.2%; P = 0.0257; skin grafting: mortality vs survival, 57.4% vs 76.2%; P = 0.0134). Multivariate regression analysis showed that tracheostomy group significantly presented with lower mortality risk by 65% (odds ratio [OR], 0.35; P = 0.0372), and subgroup analysis of delaminating follow-up duration showed that patients with tracheostomy had higher overall survival by 22% (90-day postburn mortality: nontracheostomy vs tracheostomy, 58.3% vs 36.3%; adjusted hazards ratio, 0.39; 95% confidence interval, 0.22-0.69; P = 0.0011), especially during postburn first 30 days (adjusted hazards ratio, 0.15; 95% confidence interval, 0.05-0.49; P = 0.0016). Total body surface area did not significantly affect survival; however, mortality risk was significantly higher in those with a larger TBSA (TBSA, ≥80%; OR, 6.48; P = 0.0022; TBSA, 60-79%; OR, 3.12; P = 0.0518; TBSA, 40-59%; OR, 1.88; P = 0.2402; TBSA, 30-39% as reference). CONCLUSIONS: For patients with major burn-induced AKI receiving RRT, tracheostomy and skin grafting may improve survival in the cases living through acute burn stage.

Inhibition of Sonic Hedgehog Signaling Suppresses Glioma Stem-Like Cells Likely Through Inducing Autophagic Cell Death.

Glioblastoma (GBM) often recurs after radio- and chemotherapies leading to poor prognosis. Glioma stem-like cells (GSCs) contribute to drug resistance and recurrence. Thus, understanding cellular mechanism underlying the growth of GSCs is critical for the treatment of GBM. Here GSCs were isolated from human U87 GBM cells with magnetic-activated cell sorting (MACS) using CD133 as a marker. The CD133+ cells highly expressed sonic hedgehog (Shh) and were capable of forming tumor spheroids in vitro and tumor in vivo. Athymic mice received intracranial injection of luciferase transduced parental and CD133+ GBM cells was utilized as orthotopic GBM model. Inhibited Shh by LDE225 delayed GBM growth in vivo, and downregulated Ptch1 and Gli1. CD133+ cell proliferation was more sensitive to inhibition by LDE225 than that of CD133- cells. Treatment with LDE225 significantly reduced CD133+-derived tumor spheroid formation. Large membranous vacuoles appeared in the LDE225-treated cells concomitant with the conversion of LC3-I to LC3-II. In addition, LDE225-induced cell death was mitigated in the presence of autophagy inhibitor 3-methyladenine (3-MA). Tumor growth was much slower in Shh shRNA-knockdown mice than in control RNA-transfected mice. Conversely, tumor growth was faster in Shh overexpressed mice. Furthermore, combination of LDE225 and rapamycin treatment resulted in additive effect on LC3-I to LC3-II conversion and reduction in cell viability. However, LDE225 did not affect the phosphorylated level of mTOR. Similarly, amiodarone, an mTOR-independent autophagy enhancer, reduced CD133+ cell viability and tumor spheroid formation in vitro and exhibited anti-tumor activity in vivo. These results suggest that Shh inhibitor induces autophagy of CD133+ cells likely through mTOR independent pathway. Targeting Shh signal pathway may overcome chemoresistance and provide a therapeutic strategy for patients with malignant gliomas.

The Predictive Factors Associated With Comorbidities for Treatment Response in Outpatients With King Classification III Diabetes Foot Ulcers.

OBJECTIVE: The aim of this study is to investigate the preexisting predictive factors associated with comorbidities for diabetic foot ulcer (DFU) in King classification III at an outpatient clinic. METHOD: This prospective study included 100 patients with DFU in King classification III treated at outpatient clinics in Chiayi Chang Gung Memorial Hospital from January 2011 to December 2011. The least follow-up time was 1 year. Medical documentations were in respect of patient's baseline characteristics, associated history, presence of comorbidities, follow-up time, and condition of wounds. Patients were divided into success group (healed or healing with wound reduction), stagnate group, and failure group (amputation or infection, need in-hospital medical service) in accordance with the treatment response of wounds. χ Test, Fisher exact test, and 1-way analysis of variance were used for variables in 3-group comparison, whereas Student t test was applied in 2-group comparison. The predictive factors with P value less than 0.1 were further investigated using the model of univariate logistic regression. RESULTS: With 3-group stratification according to treatment response-failure (n = 8), stagnate (n = 22), and success (n = 70)-the occurrence rate of retinopathy was higher in the treatment stagnate group (42.1%) than in the treatment failure (14.3%) and success groups (12.5%; P = 0.019); the rate of previous percutaneous transluminal angioplasty (PTA) history was higher in the treatment failure group (25%) than in the treatment stagnate (4.8%) and success groups (1.5%; P = 0.020). With 2-group stratification-failure (n = 8) versus nonfailure (n = 92), and success (n = 70) versus nonsuccess (n = 30)-PTA history was strongly associated with treatment failure (odds ratio [OR], 14.33; 95% confidence interval [CI], 1.71-120.32; P = 0.014), whereas retinopathy (OR, 0.21; 95% CI, 0.07-0.65; P = 0.006) was the major negative predictor for treatment success. Previous debridement met borderline significance to predict treatment nonsuccess (OR, 0.09; 95% CI, 0.01-1.01; P = 0.051). Sex, age, associated history, dyslipidemia, hypertension, coronary artery disease, cerebrovascular accident, chronic kidney disease, and end-stage renal disease and wound condition had no statistical significance. CONCLUSIONS: Previous PTA and retinopathy, which indicated preexisting severe vasculopathy, are univariate predictive factors for treatment failure and nonsuccess, respectively, in patients with King classification III DFU. With the subdivision of King classification III DFU, medical history taking and fundus examination are acceptable methods for risk screening at an outpatient clinic.

Successful salvage for the intractable massive chylous leakage in a cirrhotic patient: A case report and literature review.

RATIONALE: Iatrogenic chylous leakage (CL) is a rare but potentially life-threatening complication after neck surgery. In cirrhotic patient, the massive CL is even more intractable and extremely dangerous due to portal hypertension. PATIENTS CONCERNS: A 54-year-old liver cirrhotic patient had milky fluid leakage from left neck drainage tube after neck dissection surgery and hypopharyngeal cancer ablation. Electrolyte imbalance and shock status were reported when conservative managements and exploratory surgical repair failed to terminate the leakage. DIAGNOSIS: Massive CL up to >5 L/day was recorded on the post-operative day (POD) 9. INTERVENTIONS: A triangular-shaped pectoralis major (PM) muscle was designed to repair the lymph nodes dissected defect over left neck. OUTCOMES: After surgery, CL dramatically reduced to less than 300 mL/day in the coming day without relapses and terminated on the 8th days. After 3 months, the wound completely healed and the food conduit passage was patent without fistula. LESSONS: This report demonstrated the superiority of pectoralis major myocutaneous flap (PMMF) than the conservative treatment, local muscle flap, and radiologic or thoracoscopic duct ligation in cirrhotic patient with massive CL.

Synergistic inhibition of tumor growth by combination treatment with drugs against different subpopulations of glioblastoma cells.

BACKGROUND: Glioma stem cells (GSCs) contribute to tumor recurrence and drug resistance. This study characterizes the tumorigenesis of CD133+ cells and their sensitivity to pharmacological inhibition. METHODS: GSCs from human U87 and rat C6 glioblastoma cell lines were isolated via magnetic cell sorting using CD133 as a cancer stem cell marker. Cell proliferation was determined using the WST-1 assay. An intracranial mouse model and bioluminescence imaging were used to assess the effects of drugs on tumor growth in vivo. RESULTS: CD133+ cells expressed stem cell markers and exhibited self-renewal and enhanced tumor formation. Minocycline (Mino) was more effective in reducing the survival rate of CD133+ cells, whereas CD133- cells were more sensitive to inhibition by the signal transducer and activator of transcription 3 (STAT3) inhibitor. Inhibition of STAT3 decreased the expression of CD133+ stem cell markers. The combination of Mino and STAT3 inhibitor synergistically reduced the cell viability of glioma cells. Furthermore, this combination synergistically suppressed tumor growth in nude mice. CONCLUSION: The results suggest that concurrent targeting of different subpopulations of glioblastoma cells may be an effective therapeutic strategy for patients with malignant glioma.

[Effects of testosterone on insulin sensitivity in liver cells and related molecular mechanisms].

OBJECTIVE: To explore the regulation of insulin sensitivity in liver cells by androgen signaling. METHODS: Eleven adult female C57BL/6 mice were injected daily with testosterone (group T) for 24 weeks. And 10 control mice received sesame oil only (group Con). HepG2 liver cells were initially pretreated with different doses of testosterone (10(-9)-10(-5) mol/L ) for 0-36 h or with 10(-7) mol/L testosterone for 0-96 h followed by a stimulation of 100 nmol/L insulin for 15 min. Later HepG2 cells were pretreated with 10(-7) mol/L testosterone for 36 h followed by a stimulation of 100 nmol/L insulin for 15 min and then a restimulation of 100 nmol/L insulin for 15 min at 4 h and 6 h interval respectively. Phosphorylation and protein expression of Akt and GSK3ß in C57BL/6 mice liver tissues and HepG2 cells were analyzed by Western blot. RESULTS: The 24-week treatment of testosterone decreased the phosphorylation of Akt and GSK3ß in C57BL/6 adipose and liver tissues (43.1% ± 3.2% vs 77.1% ± 6.7%, 14.7% ± 6.7% vs 82.3% ± 2.0% respectively, P < 0.05). Pretreatment with 10(-8)-10(-6) mol/L testosterone within 36 h obviously increased the phosphorylation of Akt and GSK3ß (P < 0.05). However pretreatment with 10(-5) mol/L within 36 h or with 10(-7) mol/L for 96 h had no effect on the phosphorylation of Akt and GSK3ß compared with control group (P > 0.05). Pretreatment with 10(-7) mol/L testosterone for 36 h followed by insulin stimulation and restimulation after 6 h interval obviously decreased the phosphorylation of Akt and GSK3ß (P < 0.05). CONCLUSION: Androgen signaling may contribute to insulin resistance in liver cells.

[The effects of testosterone on the expression of inflammatory factors in 3T3-L1 adipocytes and its mechanism].

OBJECTIVE: To explore the effects of androgen upon the production of inflammatory Cultured factors in 3T3-L1 adipocytes and to investigate the mechanism at the molecular level. METHODS: pre-adipocytes from 3T3-L1 cell line were induced to differentiate into adipocytes. Mature adipocytes were treated with testosterone at a concentration of 10(-9) to 10(-15) mol/L for either short (0 to 30 min) or long (12, 24 and 48 h) treatment course. Secretion of inflammatory factors, i.e., interleukin-6 (IL-6) and monocyte chemoattractant protein-1 (MCP-1), in the culture medium were determined by enzyme-linked immunosorbent assay (ELISA). The mRNA expression of IL-6 and MCP-1 were determined by reverse transcriptase PCR. Phosphorylation of NF-KB was analyzed by Western blot with beta-actin as the reference gene. In another experiment, adipocytes were manipulated according to the same protocol except being pretreated with PDTC (inhibitor of NF-KB) for 2 h prior to the addition of testosterone. The results of two experiments were compared. RESULTS: (1) The secretion of IL-6 and MCP-1 in the culture medium were higher in the testosterone-treated groups than in the control groups (P < 0.05). The highest concentration of IL-6 and MCP-1 were observed in the group treated with 10(-5) M testosterone for 24 h. The mRNA expression of IL-6 and MCP-1 were higher in the groups treated with testosterone for 12 h, especially with testosterone of 10(-5) mol/L; (2) With a short treatment course of testosterone, more NF-kappaB were phosphorylated than in control, especially with testosterone of 10(-5) mol/L. More NF-kappaB was phosphorylated following the 12 h testosterone treatment (10(-9), 10(-7) and 10(-5) mol/L), especially with a testosterone concentration of 10(-9) and 10(-5) mol/L. (3) When pretreated by NF-kappaB inhibitor and followed by 10(-5) mol/L testosterone for 24 h, the secretion of IL-6 and MCP-1 in the culture medium decreased significantly (P < 0.01). Likewise, when pretreated by NF-kappaB inhibitor and followed by 10(-5) M testosterone for 12 h, the mRNA expression of IL-6 and MCP-1 decreased significantly. CONCLUSION: Within the certain scope, testosterone could increase the expression of inflammatory factors in adipocytes through the activation of NF-kappaB.

[Role of testosterone in tumor necrosis factor-alpha and monocyte chemotactic protein-1 expression in mouse macrophage RAW264.7 and its molecular mechanism].

OBJECTIVE: To investigate the role of androgen in TNF-alpha and MCP-1 expression in RAW264.7 macrophage and its molecular mechanism. METHODS: (1) RAW264.7 macrophage was treated with 10(-9) mol/L or 10(-7) mol/L testosterone (T) and then subjected to the measurement of: 1) the cellular expression of TNF-alpha and MCP-1 mRNA by RT-PCR; 2) the expression of TNF-alpha and MCP-1 in cell culture supernatant by ELISA; (2) The expression of phospho-NF-kappaB after treatment with T was measured by Western blot. (3) Cells were pre-incubated with 10(-4) mol/L PDTC (an inhibitor of NF-kappaB) for 1 hour, followed by T treatment. Expression of mRNA and supernatant levels of TNF-alpha and MCP-1 were measured by RT-PCR and ELISA. RESULTS: (1) 1) After a 6 h treatment with 10(-9) mol/L or 10(-7) mol/L T, the expression of TNF-alpha mRNA increased by 1.78 and 1.87 folds, MCP-1 by 1.58 and 1.66 folds respectively (all P < 0.05). 2) Incubated with both concentration of T for 6 h showed no significant changes of supernatant levels of TNF-alpha and MCP-1. After a 24 h treatment, the levels of TNF-alpha and MCP-1 increased significantly (all P < 0.05) while more significant increase was found in 10(-7) mol/L T group (P < 0.05). (2) The expression of phospho-NF-kappaB (ser276) increased significantly after cells were treated with 10(-7) mol/L T for 30 min (P < 0.05) and peaked at 60 min. (3) With 1 h PDTC pre-incubation, T (10(-9) mol/L or 10(-7) mol/L) treatment for 6 h led to a lower mRNA expression and 24 h led to lower supernatant levels of TNF-alpha and MCP-1 than those without (P < 0.05). However, both cellular and supernatant expression of TNF-alpha and MCP-1 with PDTC pre-incubation were still higher than those of blank controls (all P < 0.05, except for TNF-alpha in 10(-9) mol/L T treatment). CONCLUSION: Testosterone can increase TNF-alpha and MCP-1 expression in RAW264.7 macrophage in vitro. Activation of cellular NF-kappaB by testosterone may be one of underlying molecular mechanisms.

[Testosterone impairs insulin sensitivity in 3T3-L1 adipocytes and the related nongenomic mechanisms].

OBJECTIVE: To investigate the influence of rapid nongenomic effect of androgen on the insulin sensitivity of mature adipocytes and the molecular mechanism thereof. METHODS: Fetal mice [corrected] preadipocytes of the line 3T3-L1 were cultured to develop into mature adipocytes. 3T3-L1 adipocytes were pretreated with testosterone of the concentration of 10-9 to approximately 10(-5) mol/L for a short-time 0-30 minutes) or a long-time (24 hours). Insulin (Ins) 200 microl at the concentration 100 micromol/L and 2-deoxy [3H] glucose were added to examine the glucose uptake. Phosphorylation and protein expression of Ins receptor (InsR) and its downstream signaling molecules (Akt and GSK3beta) were analyzed by Western blotting. RESULTS: The Ins-stimulated glucose uptake after the pretreatment of testosterone for 30 min and 24 h decreased gradually in response to the increasing of the concentration of testosterone with the nadir both at the testosterone concentration of 10(-5) mol/L (both P < 0.05). The phosphorylation levels of InsR, Akt, and GSK3Pbeta were significantly down-regulated by adding of testosterone at the concentration of 10(-6) mol/L for 3-30 minutes (all P < 0.05) or by adding of testosterone at the concentrations of 10(-7) - 10(-6) mol/L for 24 hours (all P < 0.05). The protein expression levels of InsR, Akt, and GSK3Pbeta, however, were not significantly affected by testosterone treatment. CONCLUSION: Rapid nongenomic effect of androgen may contribute to the insulin resistance in adipocytes.

The involvement of PTEN in sleep deprivation-induced memory impairment in rats.

Although the underlying mechanism is not elucidated, it has been postulated repeatedly that deprivation of sleep, particularly rapid eye movement (REM) sleep, affects learning. Here we report that memory for newly acquired information is impaired after a specific period of REM sleep deprivation (REMD). Memory retrieval-induced phosphorylation of protein kinases in the rat amygdala is abrogated by REMD that is associated with an increase in the expression of a dual protein/lipid phosphatase PTEN. REMD given before training is without effect, suggesting the lack of effect on the acquisition of memory. Intra-amygdala administration of antisense but not sense or scrambled oligonucleotides for PTEN prevents REMD-induced decrease in protein phosphorylation and impairment of fear memory. Thus, REMD interferes with the process of memory retention via the activation of PTEN.