Effect evaluation of denosumab combined with curettage and bone cement reconstruction in the treatment of recurrent giant cell tumor of bone around the knee joint.

OBJECTIVE: Giant cell tumor of bone (GCTB) is a common primary bone tumor with latent malignant tendency. GCTB is prone to occur around the knee joint, and surgery is the major treatment method. There are relatively few reports on denosumab in the treatment of recurrent GCTB around the knee joint and postoperative function evaluation of patients. This research aimed to explore the appropriate surgical options for the treatment of recurrent GCTB around the knee joint. PATIENTS AND METHODS: 19 patients with recurrent GCTB around the knee joint, who were admitted to Hospital for 3 months following denosumab treatment from January 2016 to December 2019, were included as the research subjects. The prognosis was compared between patients treated with curettage combined with polymethylmethacrylate (PMMA) and those with extensive-resection replacement of tumor prosthesis (RTP). A deep learning model of Inception-v3 combined with a Faster region-based convolutional neural network (Faster-RCNN) was constructed to classify and identify X-ray images of patients. The Musculoskeletal Tumor Society (MSTS) score, short form-36 (SF-36) score, recurrence, and the rate of complications were also analyzed during the follow-up period. RESULTS: The results showed that the Inception-v3 model trained on the low-rank sparse loss function was obviously the best for X-ray image classification, and the classification and identification effect of the Faster-RCNN model was significantly better than that of the convolutional neural network (CNN), U-Net, and Fast region-based convolutional neural network (Fast-RCNN) models. During the follow-up period, the MSTS score in the PMMA group was significantly higher than that in the RTP group (p<0.05), while there was no significant difference in the SF-36 score, recurrence, and the rate of complications (p>0.05). CONCLUSIONS: The deep learning model could improve the classification and identification of the lesion location in the X-ray images of GCTB patients. Denosumab was an effective adjuvant for recurrent GCTB, and widely extensive-resection RTP could reduce the risk of local recurrence after denosumab treatment for recurrent GCTB.

[Early results of left ventricular assist device implantation for the treatment of heart failure].

Twenty-four male patients who underwent left ventricular assist device (LVAD) implantation due to advanced heart failure in Union Hospital, Fujian Medical University from June 2019 to June 2022 were retrospectively included. The age of patients was 32-61 (48.4±8.4) years. Everheat-Ⅰ, HeartCon and Corheart 6 left ventricular assist systems were used in 10, 6 and 8 cases, respectively. All patients were discharged successfully without mechanical failure, thrombosis or secondary thoracotomy for hemostasis. Early postoperative hemodynamics were significantly improved, left ventricular systolic diameter was reduced, left ventricular ejection fraction was gradually improved, and no hemolysis occurred. The patients were followed up for 3 to 39 (17.9±8.6) months, the cardiac function was restored to grade Ⅰ to Ⅱ, and the 6-minute walking test distance increased significantly. Therefore, satisfactory early results can be achieved with left ventricular assist device implantation for the treatment of heart failure.

[Efficacy and safety of percutaneous transluminal pulmonary angioplasty in patients with chronic thromboembolic pulmonary hypertension].

Objective: To explore the efficacy and safety of percutaneous transluminal pulmonary angioplasty (PTPA) in patients with chronic thromboembolic pulmonary hypertension (CTEPH). Methods: This prospective single arm study included 19 CTEPH patients (7 male, age(56.3±12.5)years) admitted to Wuhan Asia Heart Hospital from January 2017 to June 2019 and received PTPA interventional therapy. Baseline data, including age, sex, WHO functional class, 6-minute walk distance (6MWD), NT-proBNP, right heart catheterization values, were collected. Patients received single or repeated PTPA. Number of dilated vessels from each patient was analyzed, patients were followed up for 24 weeks and right heart catheterization was repeated at 24 weeks post initial PTPA. All-cause death, perioperative complications, and reperfusion pulmonary edema were reported. WHO functional class, 6MWD, NT-proBNP, right heart catheterization values were compared between baseline and at 24 weeks follow up. Results: Nineteen CTEPH patients received a total of 56 PTPA treatments. The pulmonary artery pressure (mPAP) decreased from (40.11±7.55) mmHg (1 mmHg=0.133 kPa) to (27.53±4.75) mmHg (P<0.001), and the total pulmonary resistance (TPR) decreased from (13.00±3.56) Wood U to (5.48±1.56) Wood U (P<0.001), cardiac output increased from (3.19±0.63) L/min to (5.23±0.94) L/minutes (P<0.01) at 24 weeks post PTPA. The WHO functional class improved significantly (P<0.001), 6MWD increased from (307.08±129.51) m to (428.00±112.64) m (P=0.002), the NT-proBNP decreased at 24 weeks post PTPA (P=0.002). During the follow-up period, there was no death; hemoptysis occurred in 4 patients during the operation, none of which resulted in serious adverse clinical consequences. One patient developed reperfusion pulmonary edema and recovered after treatment. Conclusion: PTPA treatment is safe and can significantly improve the hemodynamics and WHO functional class of patients with CTEPH.

[Consistency between bioelectrical impedance analysis and dual-energy X-ray absorptiometry for body composition measurement in children aged 7-17 years].

Objective: To evaluate the consistency between bioelectrical impedance analysis (BIA) and dual-energy X-ray absorptiometry (DXA) in the measurement of body composition in children and adolescents aged 7-17 years. Methods: Fat-free mass (FFM) and fat mass (FM) were measured by both BIA and DXA in 1 431 children. The consistency between the methods was evaluated by intra-class correlation coefficients (ICCs) and Bland-Altman analysis. Logarithmic transformation of both measurements was performed before Bland-Altman analysis. Results: The ICCs for FFM were 0.986 and 0.974 and ICCs for FM were 0.854 and 0.926 in boys and girls respectively. In boys, the mean ratio of FFMs by BIA and DXA was 1.04, with limits of Agreement (LoA) of 0.95-1.14, and in girls, the mean ratio of FFMs by BIA and DXA was 1.02, with the LoA of 0.90-1.15. The LoA of FFM became narrower with age in both boys and girls. Both boys and girls had the wide LoAs for FM (0.40-1.27 and 0.48-1.48, respectively). Additionally, the LoA ranges for FFM and FM narrowed with the increase of BMI level in both boys and girls. Conclusion: For all children, BIA showed good consistency with DXA for FFM, whereas significant errors occurred in FM measurement. The consistency between BIA and DXA was better for obese children than for underweight or normal-weight children.

Influences of hucMSC-exosomes on VEGF and BMP-2 expression in SNFH rats.

OBJECTIVE: To investigate the influences of human umbilical cord mesenchymal stem cell-derived exosomes (hucMSC-exosome) on steroid-induced necrosis of the femoral head (SNFH) and the expressions of vascular endothelial growth factor (VEGF) and bone morphogenetic protein-2 (BMP-2) in rats. PATIENTS AND METHODS: A total of 20 male Sprague-Dawley rats were randomly divided into SNFH group and SNFH + hucMSC-exosome group using a random number table. Prednisolone acetate (24.5 mg/kg) was injected twice a week to establish the rat model of SNFH, and hucMSC-exosome in a certain dose was additionally injected into the marrow cavity in SNFH + hucMSC-exosome group. After 3 weeks, the influences of hucMSC-exosome on the pathological changes and apoptosis of the femoral head in SNFH rats were detected via hematoxylin-eosin (H&E) staining and terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) staining. In addition, the expressions of cluster of differentiation 31 (CD31), VEGF, and BMP-2 in bone tissues in both groups were detected via immunohistochemical staining, and the messenger ribonucleic acid (mRNA) and protein expression levels of VEGF and BMP-2 in necrotic bone tissues in both groups were detected via Reverse Transcription-Polymerase Chain Reaction (RT-PCR) and Western blotting. RESULTS: The results of H&E staining revealed that the fibrous callus formation was good, the new trabecular structure was more obvious, the number of vacuum cleft declined, and there were fewer enlarged adipocytes in SNFH + hucMSC-exosome group compared with SNFH group. The results of TUNEL staining showed that the number of apoptotic cells in femoral head tissues was smaller in SNFH + hucMSC-exosome group (p<0.05). According to the results of immunohistochemistry, hucMSC-exosome could increase the expression of vascular endothelial marker CD31 in SNFH rats (p<0.05). Besides, the results of RT-PCR, immunostaining and Western blotting manifested that both the mRNA and protein levels of BMP-2 and VEGF in femoral head tissues were significantly increased in SNFH + hucMSC-exosome group (p<0.05). CONCLUSIONS: HucMSC-exosome can improve SNFH in rats, whose mechanism may be related to the up-regulation of VEGF and BMP-2 by exosomes.

[The value of the planned neoadjuvant radiotherapy or chemoradiotherapy for the non-radical resection of esophageal squamous cell carcinoma].

Objective: The role of planned neoadjuvant radiotherapy or chemoradiotherapy in the non-radical resection of esophageal squamous cell carcinoma was unclear. The study aimed to evaluate their therapeutic effect and analyze the prognostic factors. Methods: We retrospectively analyzed the clinical data of locally advanced esophageal squamous cell carcinoma who received neoadjuvant radio therapy (33 patients) and concurrent chemoradiotherapy (119 patients) from January 2004 to December 2016 in our single-institution database.The survival rates were calculated by Kaplan-Meier method. The prognostic factors were analyzed by using Log rank test and Cox proportional hazards model. Results: The median follow-up was 29.8 months. One hundred and one patients survived more than 3 years. The rates of overall survival (OS) and disease-free survival (DFS) at 3 years were 63.9% and 55.6%, respectively.The rates of complete, partial and minimal pathological response of the primary tumor were 50.3%, 38.4%, 11.3%, the corresponding 3-year OS were 75.5%, 57.4%, 27.3% (P<0.001) and 3-year DFS were 72.0%, 44.7%, 17.6% (P<0.001), respectively.The postoperative lymph node metastasis rate was 27.0%. The 3-year OS and DFS of the lymph node positive group was 45.6% and 32.8%, significantly lower than 70.8% and 63.7% of the negative group (both P<0.001). The 3-year OS and DFS of pathologic stage Ⅰ, Ⅱ, ⅢA, ⅢB and Ⅵ A were 76.2%, 57.4%, 64.7%, 35.0%, 33.3% (P<0.001) and 70.1%, 49.3%, 41.2%, 22.1%, 33.3% (P<0.001), respectively.The operation-related mortality was 3.3%. Multivariate analysis showed that chest pain, postoperative respiratory failure, pathological differentiation, more than 15 lymph node dissection and ypTNM stage were the independent prognostic factors of OS (P<0.05 for all). Conclusions: The planned neoadjuvant radiotherapy or chemoradiotherapy for the non-radical resection of advanced esophageal squamous cell carcinoma could result in favorable survival. The chest pain, postoperative respiratory failure, pathological differentiation, the number of lymph node resection and ypTNM stage are the independent prognostic factors of the prognosis of these patients.

Biological characteristics of osteoclast exosomes and their role in the osteogenic differentiation of somatic cells prior to osteogenesis.

This study aimed to investigate the biological characteristics of osteoclast exocrine bodies and their role in the differentiation of somatic cells, so as to find out the key factors involved in osteoclast exosomatic growth and osteogenesis. RANKL (Receptor Activator for Nuclear Factor-κ B Ligand) induced factor was used to induce the osteoclast differentiation of Raw 264.7 cells, and TRAP (Tartrate resistant acid phosphatase) staining was employed to identify induced cells. Ultra-filtration centrifugation was used to separate OC-exosomes from osteoclast supernatant, while Western blot was employed to detect the expression characteristics of exosomal proteins CD9 and CD63. PKH67 labeled exosomes were observed to target kusao cells, which were divided into 3 groups, i.e., the complete medium group (group A), the osteoblast induced group (group B), and the osteogenesis induced liquid + OC-exosomes group (group C). The medium was changed on the next day and after 14-day culture. Using Western blot, alizarin red staining and Von Kossa silver staining, the role of OC-exosomes in the differentiation of kusao cells was clarified. Results showed that TRAP staining showed osteoclasts as irregular and TRAP positive giant cells with a red multicore and a large volume. Microcapsule membrane structures with a uniform size were detected in osteoclast supernatant, and the expression of CD9 and CD63 proteins was confirmed by Western blot. In addition, the Western blot results showed that the expression of RUNX2 (Runt-related transcription factor 2) protein in group B was 1.254 times of that in group A and 2.636 times of that in group C. Furthermore, alizarin red staining showed that the ratios of calcium salt deposition area to the total area in group A, group B and group C were 0.208%, 3.469%, and 20.724%, respectively. Von Kossa silver staining showed that the ratios of calcium salt deposition area to the total area in group A, group B and group C were 0.064%, 2.636%, and 20.872%, respectively. To sum up, OC-exosomes can promote the osteogenic differentiation of osteoblast cells (kusao cells).

Insulin-like peptides regulate vitellogenesis and oviposition in the green lacewing, Chrysopa septempunctata.

Insulin-like peptides (ILPs) act through a conserved insulin signaling pathway and play crucial roles in insect metabolism, growth, reproduction, and aging. Application of bovine insulin is able to increase vitellogenin (Vg) mRNA and protein levels in female insects. Here, we first show that injection of bovine insulin into previtellogenic Chrysopa septempunctata female adults promoted ovarian growth, increased Vg protein abundance, elevated reproductive performance, and enhanced protease activity. These data suggested that ILPs play crucial roles in reproductive regulation of the green lacewing, C. septempunctata.

Evaluation of efficacy, safety and tolerability of Ambrisentan in Chinese adults with pulmonary arterial hypertension: a prospective open label cohort study.

BACKGROUND: Although several new drugs have been approved in recent years, pulmonary arterial hypertension (PAH) remains a rapidly progressive disease with a poor prognosis. Ambrisentan, a selective endothelin type A antagonist, has been approved for treatment of PAH. This open label study assessed the efficacy and safety of ambrisentan in Chinese subjects with PAH. METHODS: Eligible patients with PAH (World Health Organisation [WHO] functional class [FC] II orIII) were enrolled and received Ambrisentan (5 mg) once daily for a 12-week preliminary evaluation period, and a 12-week dose-adjustment period (dose titration to 10 mgallowed). Endpoints included: change from baseline in 6-Minute Walk Distance (6-MWD), N-Terminal Pro B-Type Natriuretic Peptide (NT-pro-BNP), WHO FC, Borg Dyspnoea Index (BDI), clinical worsening of PAH and incidences of adverse events (AE). RESULTS: One hundred thirty-three subjects (85 % women, mean age: 36 years) with PAH (WHOFC II or III) were enrolled and received ambrisentan (5 mg) once daily for a 12-week preliminary evaluation period, and a 12-week dose-adjustment period. Mean (SD) duration of drug exposure was 161.7 (27.13) days. Ambrisentan (average daily dose of 6.27 mg) significantly improved exercise capacity (6MWD) from baseline (mean: 377.1 m [m]) at week 12 (+53.6 m, p < 0.001) (primary endpoint). Improvement in exercise capacity was noted as early as week 4, and was sustained up to week 24 (+ 64.4 m, p < 0.001). NT-pro-BNP plasma levels decreased significantly (p < 0.001) at week 12 (-861.4 ng/L) and week 24 (-806 ng/L) from baseline (mean: 1600.7 ng/L). The WHO FC showed improvements for 44 subjects at week 12 and 51 subjects at week 24. BDI scores decreased significantly at week 12 (-0.3, p < 0.001) and week 24 (-0.2, p = 0.003) from baseline (mean: 2.5). Four patients died during the study (sudden cardiac death [n = 2], cerebral haemorrhage [n = 1], cardiac failure [n = 1]). Drug related adverse events occurred in 34.3 % of subjects; peripheral oedema (11.2 %) and flushing (8.2 %) occurred most frequently. CONCLUSION: Ambrisentan (5 and 10 mg, orally) significantly improved the exercise capacity in Chinese PAH subjects with a safety profile similar to that observed in global studies. TRIAL REGISTRATION: NCT No. (ClinicalTrials.gov): NCT01808313 ; Registration date (first time): February 28, 2013.

Decreased miR-452 expression in human colorectal cancer and its tumor suppressive function.

MicroRNA-452 (miR-452) is dysregulated in some human malignancies, and is correlated with tumor progression. However, its expression and function in human colorectal cancer (CRC) remain unclear. The aim of our study was to explore the effects of miR-452 in CRC tumorigenesis and development. Using reverse transcription quantitative real-time polymerase chain reaction, we detected miR-452 expression in CRC cell lines and primary tumor tissues. We also examined the association between miR-452 expression and clinicopathological factors. We then investigated the effects of miR-452 on the biological behavior of CRC cells. miR-452 expression was significantly downregulated in CRC compared with the adjacent noncancerous tissues. A low level of miR-452 was associated with larger tumor size, deeper invasion depth, and advanced TNM stage. Multivariate Cox regression analysis identified decreased miR-452 expression as an independent factor predicting poor prognosis for CRC patients. In addition, in vitro functional analysis showed that overexpression of miR-452 in HCT116 cells reduced cell proliferation, promoted cell apoptosis, and inhibited cell invasion and migration. These findings indicate that miR-452acts as a tumor suppressor in CRC, and would serve as a novel molecular therapeutic agent for the treatment of the disease.

A phase II trial of dose-dense (biweekly) paclitaxel plus carboplatin as neoadjuvant chemotherapy for operable breast cancer.

The aim of the present study is to investigate the efficacy and safety of dose-dense (biweekly) carboplatin and paclitaxel as a neoadjuvant treatment for operable breast cancer. Patients with previously untreated breast cancer (stages Ic-III) were treated with four cycles of paclitaxel (175 mg/m(2), intravenous drip, D1) and carboplatin (area under the curve of 5, D1). Patients with HER2+ disease simultaneously received trastuzumab (6 mg/kg initial dose with subsequent doses of 4 mg/kg biweekly). The primary endpoint was a pathologically complete response (pCR). Between January 2012 and February 2014, 110 patients were enrolled. The overall pCR rate was 35.45 % (39 of 110). The pCR rates for the different cancer subtypes were as follows: 10.53 % (2 of 19) among the patients with the luminal A subtype, 12.50 % (5 of 40) among the patients with the luminal B (HER2-) subtype, 58.33 % (14 of 24) among the patients with the luminal B (HER2+) subtype, 57.14 % (8 of 14) among the patients with the triple-negative subtype, and 76.92 % (10 of 13) among the patients with the HER2+ subtype. The patients experienced the following toxicity side effects: grade 3/4 neutropenia (N = 27, 24.55 %), grade 3/4 anemia (N = 6, 5.45 %), grade 3/4 thrombocytopenia (N = 2, 1.82 %), grade 3 alanine aminotransferase (ALT) elevation (N = 1, 0.91 %), grade 3 neuropathy (N = 3, 2.73 %), grade 3 pain (N = 2, 1.82 %), and grade 3 fatigue (N = 1, 0.91 %). In total, 19.09 % of the patients experienced treatment delay or discontinuation due to hematological toxicity, and one patient discontinued treatment due to non-hematological toxicity. Neoadjuvant biweekly paclitaxel plus carboplatin is a feasible therapy that achieved high pCR rates in patients with the HER2+, triple-negative, and luminal B (HER2+) cancer subtypes (NCT0205986).

Arabidopsis thaliana SEPALLATA3 protein prokaryotic expression and purification.

SEPALLATA3 (SEP3) can be attributed to E class gene of the ABCE model of floral organ development. In order to reveal how SEP3 proteins form polymers, and the relationship between the polymers and their biological functions, the experiments of Arabidopsis thaliana AtSEP3 protein soluble expression in vitro were performed to construct a vector of prokaryotic expression, and investigate induced expression of recombinant proteins in Escherichia coli cells. The protein soluble expression was analyzed through the aspects of different protein domains, induction time, induction temperature, etc. Different structural domains and expression conditions were screened, and 0.1% IPTG inducing at 22 oC for 15 h was estimated as an optimal expression strategy. The nickel chelating resin was used to purify the protein in size exclusion chromatography (SEC) and the results indicated that AtSEP3 protein was present in the form of tetramer.

Cloning, Prokaryotic Expression and Purification of CpfS1 Gene from Arabidopsis Thaliana.

CpfS1 Gene cloned from arabidopsis thaliana was expressed in Escherichia coli DH5α. A cDNA fragment about 320 bp was amplified from the total RNA of arabidopsis thaliana seeds by reverse transcription PCR (RT-PCR) with a pair of specific primers based on the sequences of the AtCpfS1 gene. The recombinant prokaryotic expression vector pET30a-AtCpfS1 was constructed by inserting the cDNA fragment encoding the mature peptide into the prokaryotic expression vector pET30a, and then transformed into E. coli DH5α. Sequence analysis showed that the fragment length was 346 bp containing a full coding region of 332 bp encoding 76 amino acid residues with a molecular mass of 21.5 kD. The SDS-PAGE electrophoresis analysis showed that the best expression was induced by 21oC and 3.6×10-3 mol/L IPTG, under which a relative molecular weight of 82.5 kD recombinant protein was produced. The nickel chelating resin was used to purify the protein in size exclusion chromatography (SEC) and the results indicated that AtCpfS1 protein was present in the form of tetramer.

Axillary lymph node status, adjusted for pathologic complete response in breast and axilla after neoadjuvant chemotherapy, predicts differential disease-free survival in breast cancer.

BACKGROUND: Our retrospective study in breast cancer patients evaluated whether integrating subtype and pathologic complete response (pcr) information into axillary lymph node restaging after neoadjuvant chemotherapy (nac) adds significance to its prognostic values. METHODS: Patients included in the analysis had stage ii or iii disease, with post-nac axillary lymph node dissection (alnd), without sentinel lymph node biopsy before completion of nac, with definitive subtyping data and subtype-oriented adjuvant treatments. The ypN grading system was used to restage axillary lymph node status, and ypN0 was adjusted by pcr in both breast and axilla into ypN0(pcr) and ypN0(non-pcr). Univariate and multivariate survival analyses were performed. RESULTS: Among the 301 patients analyzed, 145 had tumours that were hormone receptor-positive (hr+) and negative for the human epidermal growth factor receptor (her2-), 101 had tumours that were positive for her2 (her2+), and 55 had tumours that were triple-negative. The rate of pcr in both breast and axilla was 11.7%, 43.6%, and 25.5% respectively for the 3 subtypes. Compared with the non-pcr patients, the pcr patients had better disease-free survival (dfs) and overall survival (os): p = 0.002 for dfs and p = 0.011 for os. In non-pcr patients, dfs and os were similar in the ypN0(non-pcr) and ypN1 subgroups, and in the ypN2 and ypN3 subgroups. We therefore grouped the ypN grading results into ypN0(pcr) (n = 75), ypN0- 1(non-pcr) (n = 175), and ypN2-3 (n = 51). In those groups, the 3-year dfs was 98%, 91%, and 56%, and the 3-year os was 100%, 91%, and 82% respectively. The differences in dfs and os between those three subgroups were significant (all p < 0.05 in paired comparisons). Multivariate Cox regression showed that subtype and ypN staging adjusted by pcr were the only two independent factors predicting dfs. CONCLUSIONS: Axillary lymph node status after nac, adjusted for pcr in breast and axilla, predicts differential dfs in patients without prior sentinel lymph node biopsy.

Nucleation of hcp and fcc phases in bcc iron under uniform compression: classical molecular dynamics simulations.

By classical molecular dynamics simulations employing an embedded atom method potential, we have simulated the bcc to hcp/fcc structural transition in single-crystal iron under uniform compression. Results showed that the transition pressure is different from uniaxial compression and shock loading. The transformation occurs on a picosecond timescale and the transition time decreases along with the increase of pressure. The nucleation and growth of the hcp and fcc phases under constant pressure and temperature are analyzed in detail. The nucleation planes, all belonging to the {110}(bcc) family and parallel to the three compression directions [100], [010], and [001], have been observed. About 20% bcc atoms have transformed to fcc phase under pressure just over the critical point, and under higher pressure the fraction of the fcc phase increases steadily to exceed that of the hcp phase. We have investigated the transition mechanism of iron from initial bcc to hcp/fcc and found that the transition mainly consists of compression, rotation, and shuffle.

Loss of surface N-methyl-D-aspartate receptor proteins in mouse cortical neurones during anaesthesia induced by chloral hydrate in vivo.

BACKGROUND: Anaesthetics may target ionotropic glutamate receptors in brain cells to produce their biological actions. Membrane-bound ionotropic glutamate receptors undergo dynamic trafficking between the surface membrane and intracellular organelles. Their subcellular distribution is subject to modulation by changing synaptic inputs and determines the efficacy and strength of excitatory synapses. It has not been explored whether anaesthesia has any impact on surface glutamate receptor expression. In this study, the effect of general anaesthesia on expression of N-methyl-D-aspartate (NMDA) receptors in the surface and intracellular pools of cortical neurones was investigated in vivo. METHODS: General anaesthesia was induced by intraperitoneal injection of chloral hydrate in adult male mice. Surface protein cross-linking assays were performed to detect changes in distribution of NMDA receptor subunits (NR1, NR2A, and NR2B) in the surface and intracellular compartments of cerebral cortical neurones. RESULTS: Chloral hydrate did not alter the total amounts of NR1, NR2A, and NR2B proteins in cortical neurones. However, the drug reduced NR1 proteins in the surface pool of these neurones, and induced a proportional increase in NR1 in the intracellular pool. Similar redistribution of NR2B subunits was observed between the two distinct pools. The changes in NR1 and NR2B were rapid and remained throughout the duration of anaesthesia. NR2A proteins were not altered in the surface or intracellular pool in response to chloral hydrate. CONCLUSIONS: These data demonstrate that subcellular expression of NR1 and NR2B in cortical neurones is sensitive to anaesthesia. Chloral hydrate reduces surface-expressed NMDA receptors (specifically NR2B-containing NMDA receptors) in these neurones in vivo.

Molecular dynamics simulations of hcp/fcc nucleation and growth in bcc iron driven by uniaxial compression.

Molecular dynamics simulations have been performed to study the structural transition in bcc iron under uniaxial strain loading. We found that the transition pressures are less dependent on the crystal orientations, ∼14 GPa for [001], [011], and [111] loadings. However, the pressure interval of a mixed phase for [011] loading is much shorter than loading along other orientations. In addition, the temperature increased amplitude for [001] loading is evidently lower than other orientations. The nucleation and growth of the hcp/fcc phases, and their crystal orientation dependence, were analyzed in detail, where the atom structure was presented by the topological medium-range-order analysis. For [001] compression, the hcp structure occurs first and grows into a laminar morphology in the (011)(bcc) plane with some fcc atoms as an intermediate structure. For loading along [011] and [111] directions, both hcp and fcc structure nucleation and growth along the {110}(bcc) planes are observed; their morphology is also discussed.

Epidermal growth factor receptor double activating mutations involving both exons 19 and 21 exist in Chinese non-small cell lung cancer patients.

AIMS: It has been shown that the introduction of a second mutation into the already mutated epidermal growth factor receptor (EGFR) in non-small cell lung cancer (NSCLC) will alter the sensitivity to tyrosine kinase inhibitors (TKIs). EGFR double activating mutations involving both exons 19 and 21 were previously detected in Asian patients, but the sensitivity to TKIs had not yet been characterised. Our objective was to profile the status of EGFR double mutations in Chinese NSCLC patients and to further ascertain the biological properties. MATERIALS AND METHODS: In total, 145 NSCLC tumour samples from unselected Chinese NSCLC patients were sequenced to screen mutations in exons 18, 19 and 21 of EGFR. Five patients were detected to harbour the delE746-A750+L858R double activating mutations. Subcloning experiments were carried out, expression vectors inserted with corresponding full-length EGFR were constructed, and in vitro transient transfections were performed in 293T cells. Whole cell lysates were collected to assess the sensitivity to TKIs using immunoblotting. RESULTS: All five patients had adenocarcinoma. The frequency of double mutations was 3.4% (5/145). Three patients received and responded to gefitinib treatment. Subcloning experiments showed that all the subclones were either wild type or double mutated. At a concentration of TKIs of 0.1 microM, the autophosphorylation of the double mutant was inhibited greater than that of either single mutated EGFR. However, the difference disappeared when the concentration increased to 1 microM. CONCLUSIONS: delE746-A750+L858R double activating EGFR mutations exist in Chinese NSCLC patients and both locate on the same allele. These patients tend to respond well to TKIs and the sensitivity to TKIs of this double mutated EGFR is enhanced compared with either single mutant. Nonetheless, the alteration in downstream signal transduction of the double mutant remains to be determined.

Is delayed micro-graft hair transplantation possible?--evaluation of viabilities of hair follicles preserved in two storage media.

BACKGROUND: Preservation of hair grafts for the purpose of delayed transplantation would allow use of thousands of hair grafts at one time. OBJECTIVE: To evaluate the viabilities of hair follicle units preserved in DMEM and Ringer's solution at 0 degrees C for various periods. METHODS: Cell cultures and transplantation of hair follicle units into athymic mice were used. RESULTS: Outer root sheath cells could be cultivated in 95.8% of those follicles preserved in Ringer's solution, and in 86.7% of those preserved in DMEM culture medium (p < 0.01). In 84.5% of those preserved in Ringer's and 72.5% of those preserved in DMEM for 24 h, hair follicles grew well after implantation into an athymic mouse model (p < 0.05).