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Background: The combination of chemotherapeutic drugs with different pharmacological action has emerged as a promising therapeutic strategy in the treatment of cancers. Present study examines the antitumor potential of paclitaxel (PTX) and etoposide (ETP)-loaded PLGA nanoparticles for the treatment of osteosarcoma., Results: The resulting drug-loaded PLGA NP exhibited a nanosize dimension with uniform spherical morphology. The NP exhibited a sustained release profile for both PTX and ETP throughout the study period without any sign of initial burst release. The combinational drug-loaded PLGA NP enhanced the cytotoxic effect in MG63 and Saos-2 osteosarcoma cell lines, in comparison to either native drug alone or in cocktail combinations. Additionally, NPs showed an appreciable uptake in MG63 cells in a time-based manner. Co-delivery of anticancer drugs resulted in enhanced cell cycle arrest and cell apoptosis. The results clearly showed that combinational drugs remarkably improved the therapeutic index of chemotherapeutic drugs. The greater inhibitory effect of nanoparticle combination would be of great advantage during systemic cancer therapy., Conclusion: Taken together, our study demonstrated that PTX-ETP/PLGA NP based combination therapy holds significant potential towards the treatment of osteosarcoma.

Incorporation of norleucine in place of methionine residues during recombinant protein production in Escherichia coli is well known. Continuous feeding of methionine is commonly used in E. coli recombinant protein production processes to prevent norleucine incorporation. Although this strategy is effective in preventing norleucine incorporation, there are several disadvantages associated with continuous feeding. Continuous feeding increases the operational complexity and the overall cost of the fermentation process. In addition, the continuous feed leads to undesirable dilution of the fermentation medium possibly resulting in lower cell densities and recombinant protein yields. In this work, the genomes of three E. coli hosts were engineered by introducing chromosomal mutations that result in methionine overproduction in the cell. The recombinant protein purified from the fermentations using the methionine overproducing hosts had no norleucine incorporation. Furthermore, these studies demonstrated that the fermentations using one of the methionine overproducing hosts exhibited comparable fermentation performance as the control host in three different recombinant protein production processes., (© 2014 American Institute of Chemical Engineers.)

Background: Increasing attention is being given to the use of Chinese medicine (CM) for preventing and healing vascular complications of chronic ulcerative lesions of diabetic foot., Objectives: The purposes of this paper are to describe some benefits of CM for the treatment of diabetic foot and to provide some expert opinions based on some case studies and evidence from documented Chinese traditional medicine literature., Methods: A critical review of the literature and a case report., Results: Cumulative evidence in the literature indicate that CM preparations possess anti-inflammatory activities, antioxygenation, antibiosis, antibacterial, antiallergic and beneficial effect on the viability of fibroblasts. Case record suggested that after CM treatment the patient with Wagner Grade IV ulcers healed completely., Conclusion: It is becoming increasingly important for integrated CM and biomedicine therapy to treat diabetes-related vascular complications. The opportunities for effective CM interventions are significant, and more solid evidence is warranted to show the efficacy of CM in the treatment of diabetic foot ulcers in the near future.

Aim: To investigate the effect and mechanism of acupuncture at heterotopic acupoints on jejunal motility, particularly in pathological conditions., Methods: Jejunal motility was assessed using a manometric balloon placed in the jejunum approximately 18-20 cm downstream from the pylorus and filled with approximately 0.1 mL warm water in anesthetized normal rats or rats with diarrhea or constipation. The heterotopic acupoints including LI11 (Quchi), ST37 (Shangjuxu), BL25 (Dachangshu), and the homotopic acupoint ST25 (Tianshu), and were stimulated for 60 s by rotating acupuncture needles right and left at a frequency of 2 Hz. To determine the type of afferent fibers mediating the regulation of jejunal motility by manual acupuncture, the ipsilateral sciatic A or C fibers of ST37 were inactivated by local application of the A-fiber selective demyelination agent cobra venom or the C fiber blocker capsaicin. Methoctramine, a selective M2 receptor antagonist, was injected intravenously to identify a specific role for M2 receptors in mediating the effect of acupuncture on jejunal motility., Results: Acupuncture at heterotopic acupoints, such as LI11 and ST37, increased jejunal motility not only in normal rats, but also in rats with constipation or diarrhea. In normal rats, manual acupuncture at LI11 or ST37 enhanced jejunal pressure from 7.34 ± 0.19 cmH2O to 7.93 ± 0.20 cmH2O, an increase of 9.05% ± 0.82% (P < 0.05), and from 6.95 ± 0.14 cmH2O to 8.97 ± 0.22 cmH2O, a significant increase of 27.44% ± 1.96% (P < 0.01), respectively. In constipated rats, manual acupuncture at LI11 or ST37 increased intrajejunal pressure from 8.17 ± 0.31 cmH2O to 9.86 ± 0.36 cmH2O, an increase of 20.69% ± 2.10% (P < 0.05), and from 8.82 ± 0.28 cmH2O to 10.83 ± 0.28 cmH2O, an increase of 22.81% ± 1.46% (P < 0.05), respectively. In rats with diarrhea, MA at LI11 or ST37 increased intrajejunal pressure from 11.95 ± 0.35 cmH2O to 13.96 ± 0.39 cmH2O, an increase of 16.82% ± 2.35% (P < 0.05), and tended to increase intrajejunal pressure (from 12.42 ± 0.38 cmH2O to 13.05 ± 0.38 cmH2O, an increase of 5.07% ± 1.08%, P > 0.05), respectively. In contrast, acupuncture ST25, a homotopic acupoint, decreased not only intrajejunal pressure, but also significantly decreased frequency in normal rats and rats with constipation or diarrhea. Following demyelination of Aδ fibers, acupuncture at ST37 again augmented intrajejunal pressure to 121.48% ± 3.06% of baseline. Following capsaicin application for 24 h, acupuncture at ipsilateral ST37 increased intrajejunal pressure significantly to 106.63% ± 1.26% of basal levels when compared to measurements prior to capsaicin treatment (P < 0.05). Acupuncture at LI11, ST37, or BL25 significantly rescued methoctramine-mediated inhibition of jejunal motility amplitude from 42.83% ± 1.65% to 53.43% ± 1.95% of baseline (P < 0.05), from 45.15% ± 2.22% to 70.51% ± 2.34% of baseline (P < 0.01), and from 38.03% ± 2.34% to 70.12% ± 2.22% of baseline (P < 0.01), respectively., Conclusion: Acupuncture at heterotopic acupoints increases the amplitude of jejunal motility in rats. C fibers and M2 receptors predominantly and partially mediate the regulation of jejunal motility by acupuncture, respectively.

Single-nanoparticle detection and sizing is demonstrated using a nanofiber pair in an aqueous environment. The sizing of nanoparticles with a single radius (100 nm) and of mixed nanoparticles with different radii (100 nm and 170 nm) are both realized, and the experimental results agree well with predictions of Rayleigh-Gans scattering, by taking the inhomogeneous field distribution of the nanofibers into account., (© 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

To evaluate safety and efficacy of laparoscopy-assisted radical gastrectomy (LARG) for advanced gastric cancer patients aged 70 years or older. Clinical data were retrospectively collected from patients with IIA-IIIC gastric cancer who underwent LARG (n = 30) and open radical gastrectomy (ORG, n = 34) in Department of Gastrointestinal Surgery in the Ningbo First Hospital from January 2012 to December 2013. The mean operative time was longer in the LARG group than in the ORG group but there was no statistical difference between the two groups. The intraoperative blood loss (120 ± 52.7 ml vs 227.3 ± 146.9 ml), incidence of postoperative complication (23.0% vs 47%) were lower in the LARG group than those in the ORG group. In addition, the time to first flatus (2.9 ± 0.8 d vs 4.6 ± 1.2 d), time to first ambulation (1.2 ± 0.4 vs 4.1 ± 1.0 d), time of nasogastric intubation (2.5 ± 1.0 d vs 3.5 ± 1.4 d), and postoperative hospital stay (13.0 ± 4.2 d vs 16.9 ± 4.1 d) were significantly shorter in the LARG group than in the ORG group, respectively. No statistical difference in the number of harvested lymph nodes was noted between the two groups (30.2 ± 12.0 vs 28.1 ± 11.8, P > 0.05). LARG is safer, more effective and less invasive for the elderly patients with advanced gastric cancer.

Ultrasensitive nanoparticle detection holds great potential for early-stage diagnosis of human diseases and for environmental monitoring. In this work, we report for the first time, to our knowledge, single nanoparticle detection by monitoring the beat frequency of split-mode Raman lasers in high-Q optical microcavities. We first demonstrate this method by controllably transferring single 50-nm-radius nanoparticles to and from the cavity surface using a fiber taper. We then realize real-time detection of single nanoparticles in an aqueous environment, with a record low detection limit of 20 nm in radius, without using additional techniques for laser noise suppression. Because Raman scattering occurs in most materials under practically any pump wavelength, this Raman laser-based sensing method not only removes the need for doping the microcavity with a gain medium but also loosens the requirement of specific wavelength bands for the pump lasers, thus representing a significant step toward practical microlaser sensors.

Recombinant antibodies produced in Chinese hamster ovary (CHO) cells often exhibit a slight yellow-brown color, but the molecular basis for this color has remained elusive. We predicted that the color results from post-translational modifications on the antibody, because colored species were found to coelute with antibody products during size exclusion chromatography. Previously, modification by oxidation and advanced glycation end products (AGEs) had been shown to give rise to colored and fluorescent species whose spectral properties are in agreement with the spectral properties of CHO-derived recombinant antibodies. To test whether these modifications give rise to the color exhibited in our CHO-derived antibodies, we searched for 8 different oxidation and 28 different glycation and AGE modifications by mass spectrometry in a variety of samples exhibiting varying color intensities. Oxidation and glycation modifications correlated weakly with color in a subset of samples, but several AGEs exhibited a strong correlation with product color in all samples tested. This strong correlation with sample color was verified for a specific AGE, carboxymethyllysine, by ELISA, thus validating the mass spectrometry data. These data indicate that AGEs are at least partially responsible for the color seen in CHO-derived recombinant antibodies.

Objective: High-dose methotrexate (hdmtx) is a common therapeutic agent in the treatment of osteosarcoma. However, hdmtx is highly toxic and requires complex pharmacokinetic monitoring and leucovorin rescue. Thus, alternative therapeutic strategies are necessary. Here, we analyzed the clinical efficacy of a dia regimen (cisplatin-ifosfamide-doxorubicin) to evaluate its potential as an alternative to hdmtx-based therapy., Methods: Patients received 12 cycles of chemotherapy administered over 2 years (2 preoperative cycles and 10 postoperative cycles). Cumulative dose was the same in all cycles: cisplatin 120 mg/m(2) on day 1 of week 1, followed by ifosfamide 2.0 g/m(2) days 1-5 of week 2, and doxorubicin 20 mg/m(2) days 1-3 of week 2., Results: Between January 2004 and October 2008, 39 eligible patients (median age: 16 years) were enrolled, with 36 being evaluable for the study. Of those 36 patients, 20 (55.6%) had a good histologic response to preoperative chemotherapy (>90% tumour necrosis). The estimated 5-year rates of event-free survival (efs) and overall survival were 54.8% and 61.5% respectively., Conclusions: The results of our study suggest that, in osteosarcoma patients, the dia regimen produces an efs rate and survival outcomes comparable to those attained with hdmtx-containing regimens, with fewer adverse reactions. The dia regimen is well tolerated, and we observed a high level of patient compliance. Our results demonstrate that hdmtx-free osteosarcoma treatment regimens can be effective, warranting further investigation.

The incidence of cognitive disorders such as Alzheimer's disease continues to increase unabated. While cures for such diseases have eluded investigators, progress is being made on alleviating certain symptoms of these diseases. Mouse knockouts of the proline transporter (PROT), a high affinity Na(+)/Cl(-)-dependent transporter, indicated its potential as a novel therapeutic target for cognition improvement. Herein we report our investigation into a novel class of PROT inhibitors., (Copyright © 2014 Elsevier Ltd. All rights reserved.)

In an 11-year-old boy with osteosarcoma in the proximal tibia (type iii), 2 cycles of dia chemotherapy (cisplatin, ifosfamide, doxorubicin) were administered preoperatively while epiphysiolysis was performed. Clinical response was determined to be complete by radiography and histopathology. Marginal excision was then performed with preservation of the proximal tibial epiphysis. Metaphyseal reconstruction was performed using distraction osteogenesis. Six cycles of dia chemotherapy were administered postoperatively. Twenty months later, the patient had developed no complications and experienced full bone healing, with no limb discrepancy. In selected adolescent patients with osteosarcoma, in whom the tumour is in full contact with the epiphysis, epiphyseal preservation by epiphysiolysis and reconstruction by distraction osteogenesis can provide an excellent outcome, resulting in a stable reconstruction that functionally restores the native limb.

Background: Our previous studies have demonstrated the treatment benefits of dense cranial electroacupuncture stimulation (DCEAS), a novel brain stimulation therapy in patients with major depression, postpartum depression and obsessive-compulsive disorder. The purpose of the present study was to further evaluate the effectiveness of DCEAS combined with body acupuncture and selective serotonin reuptake inhibitors (SSRIs) in patients with post-stroke depression (PSD)., Methods: In a single-blind, randomized controlled trial, 43 patients with PSD were randomly assigned to 12 sessions of DCEAS plus SSRI plus body electroacupuncture (n = 23), or sham (non-invasive cranial electroacupuncture, n-CEA) plus SSRI plus body electroacupuncture (n = 20) for 3 sessions per week over 4 weeks. Treatment outcomes were measured using the 17-item Hamilton Depression Rating Scale (HAMD-17), the Clinical Global Impression - Severity scale (CGI-S) and Barthel Index (BI), a measure used to evaluate movement ability associated with daily self-caring activity., Results: DCEAS produced a significantly greater reduction of both HAMD-17 and CGI-S as early as week 1 and CGI-S at endpoint compared to n-CEA, but subjects of n-CEA group exhibited a significantly greater improvement on BI at week 4 than DCEAS. Incidence of adverse events was not different in the two groups., Conclusions: These results indicate that DCEAS could be effective in reducing stroke patients' depressive symptoms. Superficial electrical stimulation in n-CEA group may be beneficial in improving movement disability of stroke patients. A combination of DCEAS and body acupuncture can be considered a treatment option for neuropsychiatric sequelae of stroke., Trial Registration: http://www.clinicaltrials.gov, NCT01174394.

Cationic oligomers with a rigid aromatic backbone were first applied as non-viral gene delivery vectors. These materials showed better DNA condensation ability than their flexible analogues. In vitro transfection experiments revealed that the materials with more rigid backbone exhibited considerably higher TE and lower cytotoxicity than 25 kDa PEI.

Recent genetic evidence suggests that the diacylglycerol lipase (DAGL-α) isoform is the major biosynthetic enzyme for the most abundant endocannabinoid, 2-arachidonoyl-glycerol (2-AG), in the central nervous system. Revelation of its essential role in regulating retrograde synaptic plasticity and adult neurogenesis has made it an attractive therapeutic target. Therefore, it has become apparent that selective inhibition of DAGL-α enzyme activity with a small molecule could be a strategy for the development of novel therapies for the treatment of disease indications such as depression, anxiety, pain, and cognition. In this report, the authors present the identification of small-molecule inhibitor chemotypes of DAGL-α, which were selective (≥10-fold) against two other lipases, pancreatic lipase and monoacylglycerol lipase, via high-throughput screening of a diverse compound collection. Seven chemotypes of interest from a list of 185 structural clusters, which included 132 singletons, were initially selected for evaluation and characterization. Selection was based on potency, selectivity, and chemical tractability. One of the chemotypes, the glycine sulfonamide series, was prioritized as an initial lead for further medicinal chemistry optimization.

Objective: To observe the cardioprotective effect of electroacupuncture (EA) stimulation at "Neiguan" (PC 6) in mice with myocardial ischemia (MI) and to explore the involvement of cardiac beta-adrenergic receptors (beta-AR) in the cardioprotective effect of EA intervention., Methods: Adult male C 57 BL/6 mice of wild type and beta1/beta2-AR double-knockout (beta1/beta2-AR(-/-)) mice were randomly and respectively divided into control group and EA group, with 8 mice in each group. These mice were subject to procedures of basic control, fatigue swimming and fatigue swimming + EA. The model was established by fatigue swimming for inducing acute MI. EA (2 Hz, 0.5 mA) was applied to bilateral "Neiguan" (PC 6) for 30 min, once daily for 7 days in both EA groups. The ST-segment of electrocardiogram (ECG) of standard limb lead II, heart rate were recorded by using a biophysical amplifier and the arrhythmia score was assessed according to Curtis and Walker's methods (1988)., Results: Self-comparison showed that, compared with the baseline, the amplitude of ECG-ST II segment was obviously increased in swimming fatigue C 57 BL/6 mice (P < 0.01) and further obviously increased in beta1/beta2-AR(-/-) mice (P < 0.01), suggesting an acute MI in C 57 BL/6 mice and a worsened MI in beta1/beta2-AR(-/-) mice. Simultaneously, the heart rate was markedly decreased (P < 0.05), and the score of arrhythmia obviously increased in both C 57 BL/6 and beta1/beta2-AR(-/-) mice (P < 0.05). Compared with the modeling procedures, ECG-ST II amplitude and arrhythmia score were significantly decreased in C 57 BL/6 mice of the EA group (P < 0.01, P < 0.05), rather than in the beta1/beta2-AR(-/-) mice of the EA group (P > 0.05). In addition, heart rate levels of both C 57 BL/6 mice and beta1/beta2-AR(-/-) mice had no significant differences between control and EA groups (P > 0.05)., Conclusion: EA at "Neiguan" (PC 6) can protect the heart from swimming fatigue-induced MI in C 57 BL/6 mice not in beta1/beta2-AR(-/-) mice, suggesting an involvement of beta1/beta2-AR in the protective effect of EA.

We aimed to investigate the antitumor effect and mechanism of fructose 1,6-bisphosphate (F1,6BP) in a hepatocellular carcinoma cell line. HepG2 cells were treated with different concentrations of F1,6BP alone or in combination with antioxidant N-acetyl-L-cysteine (NAC) or catalase (CAT), and cell proliferation assays were performed. Nuclear morphology was observed by fluorescence microscopy after Hoechst staining, and apoptosis was measured with flow cytometry. Changes in reactive oxygen species (ROS) levels in HepG2 cells were detected by 2',7'-dichlorodihydrofluorescein diacetate (DCFH-DA) staining. A colorimetric assay was adopted to determine the percentage of oxidized glutathione in these cells. CAT and glutathione peroxidase (GSH-Px) mRNA expression levels in HepG2 cells were measured by real-time fluorescence quantitative PCR. HepG2 cell proliferation was significantly inhibited by F1,6BP, accompanied by an increase in intracellular ROS levels and oxidized glutathione. Upregulated apoptosis and characteristic nuclear morphological changes were observed, and the expression of CAT and GSH-Px mRNA was increased after F1,6BP treatment. The antitumor effect of F1,6BP was significantly decreased after pretreatment with NAC and CAT in HepG2 cells. In conclusion, F1,6BP can induce the apoptosis of HepG2 cells. The mechanism involved may be associated with the generation of ROS, especially the production of H2O2.

An operative approach to the inferior pubic ramus that was utilized in four patients with various bone tumors in the inferior pubic ramus is described here. These patients were successfully managed though a femoribus internus (inner thigh)-perineal approach. Data concerning preoperative and postoperative symptoms, surgical procedures, and outcomes are presented. There was no recurrence in the four cases and the pain associated with an initial pelvic floor had completely resolved except one case. The slight limitation in range of motion of the left hip joint and pain were performed in the same case postoperatively. The Musculoskeletal Tumor Society scores were 28, 15, 25, and 18 at the final follow-up. A typical case is described in full and our experience concerning surgical indications, and intraoperative issues in tumor patients discussed. The purpose of this paper is to recommend that the femoribus internus-perineal approach be used to resect the inferior pubic ramus, whether affected by osteomyelitis, bone tumor, or tuberculosis, but especially in patients with tumors., (© 2014 Chinese Orthopaedic Association and Wiley Publishing Asia Pty Ltd.)

Aim: To evaluate clinical outcomes in patients who underwent alcohol-inactivated autograft replantation with joint preservation for the treatment of osteosarcoma of the distal femur., Patients and Methods: 10 patients (7 men, 3 women; mean age: 20.1 years) were treated. 9 patients had Enneking stage IIb tumors and 1 patient had stage III. The mean follow-up time was 34 months. All patients obtained first-stage healing., Results: 1 patient died of local recurrence and multiple metastases 13 months after surgery; 3 patients died of multiple metastases 9, 12, and 24 months after surgery, respectively. 3 patients required second surgery because of fracture of the inactivated autograft, 1 patient died 6 months later, 1 patient experienced bony healing 4 months later with no further complications, and 1 patient required subsequent open reduction and bone implantation with internal fixation (bony healing was evident 6 months later; however, joint instability and limited knee flexion were apparent). At final follow-up, the mean International Society of Limb Salvage (ISOLS) graft score was 31 (87%), whereas the mean Musculoskeletal Tumour Society (MSTS) function score was 23 (77%)., Conclusions: These preliminary findings indicate that alcohol-inactivated autograft replantation with joint preservation may be effective in treating osteosarcoma of the distal femur in some patients. Larger-scale studies are needed to fully evaluate the efficacy/safety of this approach., (© 2014 S. Karger GmbH, Freiburg.)

Objective: This study tested multiple dosing epochs of pre-loaded D-methionine (D-met) for otoprotection from noise-induced hearing loss (NIHL)., Design: Auditory brainstem response (ABR) thresholds were measured at baseline, 1 day, and 21 days following a 6-hour 105 dB sound pressure level (SPL) octave band noise (OBN) exposure. Outer hair cell (OHC) counts were measured after day 21 sacrifice., Study Sample: Three groups of five Chinchillas laniger each were given a 2-day regimen comprising five doses of D-met (200 mg/kg/dose) intraperitoneally (IP) starting 2, 2.5, or 3 days prior to noise exposure. A control group (n = 5) received five doses of equivalent volume saline IP starting 2.5 days prior to noise exposure., Results: ABR threshold shifts from baseline to day-21 post-noise exposure were reduced in all D-met groups versus controls, reaching significance (p < 0.05) in the 3-day group. D-met groups showed reduced OHC loss relative to controls at day-21 post-noise exposure, reaching significance (p < 0.05) at all frequency regions in the 3-day group and at the 2, 4, and 8 kHz frequency regions in the 2.5-day group., Conclusions: D-met administration in advance of noise-exposure, without further administration, significantly protects from noise-induced ABR threshold shift and OHC loss.

A new label-free sensing mechanism is demonstrated experimentally by monitoring the whispering-gallery mode broadening in microcavities. It is immune to both noise from the probe laser and environmental disturbances, and is able to remove the strict requirement for ultra-high-Q mode cavities for sensitive nanoparticle detection. This ability to sense nanoscale objects and biological analytes is particularly crucial for wide applications.

Giant cell tumour of bone (gctb) is one type of giant-cell-rich bone lesion characterized by the presence of numerous multinucleated osteoclast-type giant cells. Giant cells are known to express rankl (receptor activator of nuclear factor κB ligand) and are responsible for the aggressive osteolytic nature of the tumour. No available treatment option is definitively effective in curing this disease, especially in surgically unsalvageable cases. In recent years, several studies of denosumab in patients with advanced or unresectable gctb have shown objective changes in tumour composition, reduced bony destruction, and clinical benefit. Denosumab is a fully human monoclonal antibody that targets and binds with high affinity and specificity to rankl. Several large phase iii studies have shown that denosumab is more effective than bisphosphonates in reducing skeletal morbidity arising from a wide range of tumours and that it can delay bone metastasis. The relevant articles are reviewed here. The controversies related to the future use of denosumab in the treatment of gctb are discussed.

This paper studied the changes of organic soil substrate properties with increasing cultivation years and their effects on the cucumber growth in solar greenhouse. The results showed that with increasing cultivation years, the physical and chemical properties of organic soil substrate deteriorated, which was manifested in the increase of bulk density and the decrease of total porosity, pH, and available nutrient contents. The numbers of bacteria and actinomycetes in the substrate decreased with increasing cultivation years, while that of fungi was in reverse. The cucumber growth was also affected to a certain extent, manifesting in the decrease of plant height and leaf area, the decline of photosynthetic efficiency, and the decrease of yield and quality. It would be necessary to restore the fertility of organic soil substrate after 3-year cultivation.

We demonstrate that the nonlinear optomechanical interaction leads to parametric down-conversion, capable of generating polariton pairs formed by photons and phonons. The nonlinearity is resonantly enhanced through frequency matching, and such parametric down-conversion does not require the stringent condition that the single-photon optomechanical coupling strength g be on the order of the mechanical resonance frequency ω(m). We provide analytical results for the frequency matching condition and derive the nonlinear coefficient. Numerical simulations on polariton pair generation are presented, showing that photonlike polaritons, phononlike polaritons, and mixed photon-phonon polaritons can be selectively generated. Such nonlinear interaction offers a promising way for harnessing the optomechanical nonlinearity to manipulate photons and phonons.

The interaction between jatrorrhizine (JAT) and bovine serum albumin (BSA) has been studied. The studies were carried out in a buffer medium at pH 7.4 using fluorescence spectroscopy, UV-vis spectroscopy, and molecular modeling methods. The results of fluorescence quenching and UV-vis absorption spectra experiments indicated the formation of the complex of BSA-JAT. Binding parameters were determined using the Stern-Volmer equation and Scatchard equation. The results of thermodynamic parameters ΔG, ΔH and ΔS at different temperatures indicate that the electrostatic interactions and hydrogen bonds play a major role for JAT-BSA association. Site marker competitive displacement experiments and molecular modeling calculation demonstrating that JAT is mainly located within the hydrophobic pocket of the subdomain IIIA of BSA. Furthermore, The distance between donor (BSA) and acceptor (JAT) was estimated according to fluorescence resonance energy transfer.

Objective: To conduct a systematic review and meta-analysis of the performance of the procalcitonin (PCT) diagnostic test for identifying infectious complications after hematopoietic stem cell transplantation (HSCT)., Methods: We searched EMBASE, MEDLINE, the Cochrane database, and reference lists of relevant articles, with no language restrictions, through December 2011. We selected original articles that reported diagnostic performance of PCT alone or compared with other biomarkers for identifying serious infections in HSCT recipients. We quantitatively evaluated test accuracy parameters with the use of forest plots, hierarchical summary receiver operating characteristic curves, and bivariate random effect models., Results: We found 6 qualifying studies (studying 1344 episodes of suspected infection with confirmed infectious episodes) from 3 countries. These 6 studies examined both PCT and C-reactive protein (CRP) test performance. Bivariate pooled sensitivity, specificity, positive likelihood ratios, and negative likelihood ratios were 0.66 (95% confidence interval [CI] 0.60-0.72), 0.72 (95% CI 0.65-0.79), 2.39 (95% CI 1.84- 3.09), and 0.47 (95% CI 0.39-0.57) for PCT, and 0.80 (95% CI 0.54-0.93), 0.73 (95% CI 0.56-0.86), 3.00 (95% CI 1.86-4.84), and 0.27 (95% CI 0.11-0.65) for CRP. In terms of area under the curve (AUC), CRP was superior to PCT in detecting infectious complications, with an AUC of 0.82 for CRP versus an AUC of 0.69 for PCT., Conclusion: The pooled accuracy estimates of 6 different studies indicated only a moderate rule-out diagnostic value of both PCT and CRP in discriminating infection from other inflammatory complications following allogeneic HSCT., (© 2013 John Wiley & Sons A/S.)

Amino acid sequence variants are defined as unintended amino acid sequence changes that contribute to product variation with potential impact to product safety, immunogenicity, and efficacy. Therefore, it is important to understand the propensity for sequence variant (SV) formation during the production of recombinant proteins for therapeutic use. During the development of clinical therapeutic products, several monoclonal antibodies (mAbs) produced from Chinese Hamster Ovary (CHO) cells exhibited SVs at low levels (≤3%) in multiple locations throughout the mAbs. In these examples, the cell culture process depleted tyrosine, and the tyrosine residues in the recombinant mAbs were replaced with phenylalanine or histidine. In this work, it is demonstrated that tyrosine supplementation eliminated the tyrosine SVs, while early tyrosine starvation significantly increased the SV level in all mAbs tested. Additionally, it was determined that phenylalanine is the amino acid preferentially misincorporated in the absence of tyrosine over histidine, with no other amino acid misincorporated in the absence of tyrosine, phenylalanine, and histidine. The data support that the tyrosine SVs are due to mistranslation and not DNA mutation, most likely due to tRNA(Tyr) mischarging due to the structural similarities between tyrosine and phenylalanine., (Copyright © 2012 Wiley Periodicals, Inc.)

Objective: To compare the effects of carbamyl-beta-methylcholine chloride (CMCC) administered by intra-acupuncture point injection (IAI), intramuscular injection (IMI), and intravenous injection (IVI), and to analyze the mechanisms., Methods: In the IAI group, CMCC was injected into the Zusanli acupoint (ST 36) immediately after 30-min stimulation by electro-acupuncture (EA) at the acupoints, and into the femoral vein and skeletal muscle in IVI and IMI groups, respectively. Intra-gastric pressure was detected. The plasma concentration of CMCC was measured at various times., Results: The gastric effect of CMCC in the IVI group was enhanced and attenuated more rapidly than in the other groups. In the IAI group, this effect was significantly stronger than that in the IMI group at 2 min and 15 min, but not significantly different between the two groups at 5 min and 30 min. Plasma concentration of CMCC in the IAI group was similar to that in the IVI group at 2 min, but higher than that in the IMI group. The concentration in the IAI group was higher than that in the IV group and similar to that in the IMI group at 5, 15 and 30 min, indicating rapid increase and slower reduction of the plasma concentration of the drug in the IAI group. There was a positive correlation between the plasma concentration of CMCC and intragastric pressure in all groups., Conclusion: The effect of IAI with CMCC was stronger than that of IMI and longer-lasting than that of IVI, which correlated with the blood concentration of CMCC.

The aim of the present study was to investigate the apoptosis of human ovarian cancer cell lines, A2780 and CP70, induced by a novel curcumin analogue, B19. The proliferation of cells was detected with methyl thiazolyl tetrazolium (MTT) assay and apoptosis was examined by flow cytometry. Reactive oxygen species (ROS) were assessed by the fluorescent indicator DCF-DA. The protein expression of the endoplasmic reticulum (ER) stress pathways, GRP78, XBP-1, ATF-4 and CHOP, was examined with Western blotting. A growth inhibitory effect was observed after treatment with B19 in a dose-dependent manner and with more potential than curcumin. At 20 µM, B19 induced significant apoptosis in CP70 cells. Furthermore, B19 induced the ER stress response, while curcumin had no effect on ER stress. These results suggest that B19 has more effective antitumor properties than curcumin, and is associated with the activation of ER stress and ROS in ovarian cancer cells.

Objectives: The pharmacokinetics and tissue distribution of icariin propylene glycol-liposome suspension (ICA-PG-liposomes) have been investigated., Methods: ICA-PG-liposomes or ICA-PG-solution were prepared and intraperitoneally injected to mice. Morphology and size distribution of ICA-PG-liposomes were observed by transmission electron microscopy (TEM) and laser particle sizer. Plasma and tissues were collected at different times after intraperitoneal injection and icariin concentrations were determined by HPLC., Key Findings: From TEM, ICA-PG-liposomes showed spherical vesicles with a mean particle size of 182.4 nm. The encapsulation efficiency of ICA-PG-liposomes reached 92.6%. Pharmacokinetics of ICA-PG-liposomes displayed the three open compartments model. ICA-PG-liposomes enhanced icariin absorption from the abdominal cavity, prolonged mean retention time (MRT((0-t))), increased area under curve (AUC((0-t))) and maximum concentration in plasma. Compared with ICA-PG-solution, ICA-PG-liposomes resulted in larger amounts of icariin being distributed into spleen (60.38% total icariin), liver (16.68%), lung (6.21%), kidney (4.64%), heart (1.43%) and brain (1.83%). AUC((0-t)) values in most tissues (except lung) of mice administered ICA-PG-liposomes were higher than those administered ICA-PG-solution, while Clearance in most tissues (except brain and lung) decreased. The MRT((0-t)) values of ICA-PG-liposomes in all tissues and half lives of most tissues (except brain) were prolonged. From Targeted efficiency and relative uptake data, the spleen was the target tissue of the ICA-PG-liposomes., Conclusions: ICA-PG-liposomes changed the pharmacokinetic behaviour and enhanced icariin distribution in tissues. With nanometer size, high encapsulation efficiency and improved pharmacokinetics, ICA-PG-liposomes might be developed as promising carriers for icariin injection., (© 2011 The Authors. JPP © 2011 Royal Pharmaceutical Society.)

Objectives: To study the long-term outcomes of inactivated bone reimplantation with preservation of the epiphysis in children with distal femoral osteosarcomas., Methods: Over 10 years, five children (mean age 9.2 years, one boy and four girls) with distal femoral osteosarcomas underwent inactivated bone reimplantation with preservation of the epiphysis following chemotherapy in our hospital. Three patients were type I on MRI classification (one with pathological fracture), and two type II. The therapeutic regime was two cycles of preoperative chemotherapy, surgery and six cycles of postoperative chemotherapy., Results: Five patients were followed up for 60-126 months (mean 82 months). No local tumor recurrences or metastases occurred. Three patients with fractures of inactivated bone were treated by open reduction, bone grafting and internal fixation; their fractures had united by 6 months after reoperation. The mean functional score of the affected limbs was 25.6 points (13-30 points)., Conclusions: Inactivated bone reimplantation with preservation of the epiphysis for distal femoral osteosarcomas in children optimizes recovery of limb function and preservation of limb length. The main measures for improving clinical outcomes include preoperative analysis of the lesion's boundaries and extent of tumor invasion, bone grafting between inactivated and host bone, and timely treatment of complications., (© 2012 Tianjin Hospital and Blackwell Publishing Asia Pty Ltd.)

By adopting organic substrate culture, and salt-sensitive cucumber variety 'Jinchun No. 2' was used as test material, this paper studied the effects of inoculating arbuscular mycorrhiza fungi (AMF) on the plant growth, fruit yield, and fruit quality of cucumber under salt stress. AMF-inoculation could effectively promote the plant growth and nutrient uptake, and improve the fruit yield and fruit nutrient quality, compared with ordinary cultivation. Under salt stress, the plant growth was inhibited, and the plant N, P, K, Cu, and Zn contents and K+/Na+ ratio, fruit yield, and fruit soluble protein, total sugar, vitamin C, and nitrate contents decreased, while inoculation with AMF could mitigate the inhibitory effect of salt stress on the plant growth, made the plant N, P, K, Cu, and Zn contents increased by 7.3%, 11.7%, 28.2%, 13.5%, and 9.9%, respectively, and made the plant K+/Na+ ratio, fruit yield, and fruit soluble protein, total sugar, and vitamin C contents have an obvious increase and the fruit nitrate content have a significant decrease. It was suggested that AMF could promote the plant growth and nutrient uptake of cucumber under salt stress, increase the plant salt-tolerance, and improve the fruit yield and its nutrient quality.

Diacylglycerol lipase α is the key enzyme in the formation of the most prevalent endocannabinoid, 2-arachidonoylglycerol in the brain. In this study we identified the catalytic triad of diacylglycerol lipase α, consisting of serine 472, aspartate 524 and histidine 650. A truncated version of diacylglycerol lipase α, spanning residues 1-687 retains complete catalytic activity suggesting that the C-terminal domain is not required for catalysis. We also report the discovery and the characterization of fluorogenic and chromogenic substrates for diacylglycerol lipase α. Assays performed with these substrates demonstrate equipotent inhibition of diacylglycerol lipase α by tetrahydrolipastatin and RHC-20867 as compared to reactions performed with the native diacylglycerol substrate. Thus, confirming the utility of assays using these substrates for identification and kinetic characterization of inhibitors from pharmaceutical collections., (Copyright © 2011 Elsevier Inc. All rights reserved.)

We report an efficient, high fidelity trypsin digestion method for peptide map analysis. This method minimizes artifacts caused by the sample preparation process, and we show its utility for the accurate determination of succinimide formation in a degraded monoclonal antibody product. A basic charge variant was detected by imaged capillary isoelectric focusing and was shown with reduced antigen binding and biological activity. Samples were reduced under denaturing conditions at pH 5.0, and digestion of the reduced protein with porcine trypsin was performed at pH 7.0 for 1 h. Following reversed phase high-performance liquid chromatography and online mass spectrometric analysis, succinimide formation was identified at Asp30 in the light chain. This result contrasts with the observation of only iso-Asp and Asp residues under conventional sample preparation conditions, which are therefore concluded to be artificially generated. The Asp30 residue is seen in the cocrystal structure model to participate in favorable charge interaction with an antigen molecule. Formation of succinimide and the resulting loss of negative charge are therefore hypothesized to be the degradation mechanism. After treatment of the degraded antibody sample to mildly alkaline pH conditions, we observed only Asp residue as the succinimide hydrolysis product and concurrent recovery of biological activity.

Taking the energy-saving solar greenhouses with the same infrastructure but different sunken depths (0, 0.5, 1.0, and 1.5 m) in Tai' an of Shandong Province as test objects, this paper analyzed the intercepted amount of direct solar radiation energy, and studied the diurnal variation and spatial distribution patterns of environmental factors in the greenhouses on winter solstice (December 20-24, 2009) and summer solstice (June 19-23, 2010). With the increase of sunken depth, the shadow areas in the greenhouses caused by sunken profiles increased gradually, the direct solar radiation energy into the interior of the greenhouses shifted from south to north, and the ratio of ground radiation to back wall radiation decreased gradually. Within the range of 0-1.0 m sunken depth, the air temperature and soil temperature in the greenhouses increased significantly with increasing sunken depth; but when the sunken depth was 1.5 m, the warming effect declined significantly, and the deviation of the lowest soil temperature increased. The deeper the sunken depth, the lower the light intensity and the higher the relative humidity in the greenhouses were. In considering of both lighting and heat preservation, the appropriate sunken depth of energy-saving sunlight greenhouses with a span of 10 m in Tai' an region should be less than 1.0 m.

Digitalis glycosides, a group of cardiotonic agents for heart failure, have been used for a long time, but may often trigger arrhythmias as a result of digitalis intoxication. It is of great significance in finding a method to reduce their toxicity and improve clinical curative effects simultaneously in the application of digitalis glycosides. It has been well documented that acupuncture has good therapeutic effects in improving cardiac function and inhibiting arrhythmia induced by myocardial ischemia. Acupuncture combined with chemotherapeutics can reduce the adverse effects of chemotherapeutics. Accordingly, acupuncture combined with digitalis is likely to reduce the side effects of digitalis by regulating intracellular Ca2+, improving the function of Ca(2+)-ATPase in the sarcoplasmic reticulum, increasing calcium sensitivity of cardiac troponin, etc. These considerations may provide a novel clue for treatment of heart failure by joint administration of acupuncture and drugs.

GDP-mannose pyrophosphorylase (GMPase: EC 2.7.7.22) plays a crucial role in the synthesis of L-ascorbate (AsA) and the consequent detoxification of reactive oxygen species (ROS). Herein, a GMPase (accession ID DQ449030) was identified and cloned from tomato. The full-length cDNA sequence of this gene contains 1,498 bp nucleotides encoding a putative protein with 361 amino acid residues of approximate molecular weight 43 kDa. Northern blot analysis revealed that the GMPase was expressed in all examined tomato tissues, but its expression level was up-regulated in tomato plants subjected to abnormal temperatures. We then overexpressed this tomato GMPase in tobacco plants and observed that the activity of GMPase and the content of AsA were significantly increased by two- to fourfold in the leaves of transgenic tobacco plants. The effect of this gene overexpression was superimposed by the treatments of high or low temperature in tobacco, since the activities of both chloroplastic SOD (superoxide dismutase EC 1.15.1.1), APX (ascorbate peroxidase EC 1.11.1.7) and the content of AsA in leaves were significantly higher in transgenic plants than those of WT, while the contents of H(2)O(2) and O(2)(-·) were reduced. Meanwhile, relative electric conductivity increased less in transgenic plants than that in WT, and the net photosynthetic rate (P(n)) and the maximal photochemical efficiency of PSII (F(v)/F(m)) of transgenic plants were notably higher than those of WT under temperature stresses. In conclusion, the overexpression of GMPase increased the content of AsA, thereby leading to the increase in tolerance to temperature stress in transgenic plants.

Objective: To study the curative effects of pirfenidone (PF) on pulmonary fibrosis induced by paraquat (PQ) in mice and to provide the theoretical basis for clinical treatment., Methods: Ninety adult healthy male ICR mice were randomly divided into six groups: control group, PQ group, 2 mg/kg Dexamethasone group, 25 mg/kg PF group, 50 mg/kg PF group and 100 mg/kg PF group, there were 15 mice in each group. The corresponding volume of normal saline was given to the each mouse in control group according to the weight, after 2 h 0.1% CMC was given to the each mouse of control group one time by intragastric administration, then the CMC was administrated at regular time until sacrifice. All mice for other 5 groups were exposed to 100 mg/kg PQ by intragastric administration. At 2 h after exposure to PQ, 0.02 ml/10 g dexamethasone and 25, 50, 100 mg/kg PF were given to mice for dexamethasone group and for 3 PF groups by intragastric administration each day for 49 days, respectively. The lung coefficient was calculated and pathological changes of lung tissue were observed by HE staining for each mouse. The hydroxyproline (HYP) level in lung tissue was measured for each mouse. The mRNA level of and the protein level of TGF-β(1) in lung tissue for each mouse were determined, and the protein level of TGF-β(1) in the bronchus-alveolus lavage fluid (BALF) of each mouse was detected., Results: The survival rates on the 3rd day in PQ group, 3 PF groups and dexamethasone group were 53.33%, 46.67%, 73.33%, 86.67% and 80%, respectively. The survival rates on the 3rd day in dexamethasone group, 50 mg/kg and 100 mg/kg PF groups were significantly higher than those of PQ group and 25 mg/kg PF group (P < 0.05). The lung coefficients of 3 PF groups were significantly lower than that of the PQ group (P < 0.05). The lung tissue HYP levels of dexamethasone group and 3 PF groups were 50.95 ± 11.65, 44.52 ± 9.48, 43.27 ± 6.01 and 40.82 ± 5.90 mg/g respectively, which were significantly lower than that (74.27 ± 3.68) of PQ group (P < 0.01). The TGF-β(1) protein levels of BALF in dexamethasone group, 50 and 100 mg/kg PF groups were 22.03 ± 7.27, 27.75 ± 5.84 and 21.31 ± 6.82 ng/ml respectively, which were significantly lower than that (52.52 ± 15.51) ng/ml of PQ group (P < 0.01) The expression level of TGF-β(1) mRNA in 100 mg/kg PF group decreased significantly, as compared with PQ group (P < 0.01)., Conclusion: PF could reduce the collagen deposition and pulmonary fibrosis induced by PQ in mice lungs.

Objective: To optimize the preparation technics of wumeitougu oral liquid (WTOL) by response surface methodology., Methods: Based on the single-factor tests, the times of WTOL extraction, alcohol precipitation concentration and pH value were selected as three factors for box-behnken central composite design. The response surface methodology was used to optimize the parameters of the preparation., Results: Under the condition of extraction time 1.5 h, extraction times 2.772, the relative density 1.12, alcohol precipitation concentration 68.704%, and pH value 5.0, he theory highest content of Asperosaponin VI was up to 549.908 mg/L. Considering the actual situation, the conditions were amended to three extract times, alcohol precipitation concentration 69%, pH value 5.0, and the content of Dipsacaceae VI saponin examined was 548.63 mg/L which was closed to the theoretical value., Conclusion: The optimized preparation technics of WTOL by response surface methodology is reasonable and feasible.

Objective: To explore the relationship between the effect of electroacupuncture (EA) and EA of different layer tissues of the acupoint area and different acupoints in upregulating mean arterial pressure (MAP) and heart rate (HR) in hypotension plus bradycardia rats., Methods: A total of 200 SD rats were used in the present study. Bradycardia plus hypotension model was established by intravenous injection of 0.4% propranolol (0.4 mg/100 g, maintaining dosage 0.025 mg/100 g per minute). EA (2 Hz/15 Hz, 5 mA) was applied to (1) right "Daling" (PC7) and "Jiexi" (ST 41), "Ximen" (PC 4) and "Housanli" (ST 36), "Quze" (PC 3) and "Dubi" (ST 35) which have a similar tissue structure, and are located in the upper and lower limbs and different meridians, and non-acupoint [3 mm left-superior to the "Tianshu" (ST 25)], (2) skin, muscle layer and periosteum part of "Ximen" (PC4), (3) skin, muscle layer and periosteum of "Housanli" (ST36) for 15 min. The HR and MAP were recorded by using a multi-channel physiological signal sampling-processing system., Results: (1) In comparison with the model group, the percentages of the increased HR and MAP in the "Ximen" (PC4), "Quze" (PC3), "Housanli" (ST 36) and "Jiexi" (ST41) groups, PC 4-skin, PC 4-muscle, PC 4-periosteum, ST 36-skin, ST 36-muscle and ST 36 periosteum groups, and the increased HR in the "Dubi" (ST 35) group were upregulated significantly (P < 0.05, P < 0.01). The percentages of the increased HR and MAP were significantly higher in the "Quze" (PC3) and "Ximen" (PC4) groups than in the "Daling" (PC7) group (P < 0.01), and the increased HR evidently higher in the "Housanli" (ST36) and "Jiexi" (ST41) groups than in the "Dubi" (ST35) group (P < 0.01), suggesting different effects of EA stimulation of different acupoints in the same one meridian. No significant differences were found among the "Ximen" (PC4), "Quze" (PC3), "Housanli" (ST36) and "Jiexi" (ST 41) groups, and between the "Daling" (PC7) and model groups, and between the non-acupoint and model groups in the rising rates of both HR and MAP (P > 0.05). (2) Regarding the effects of EA of different tissue layers in "Ximen" (PC4) and "Housanli" (ST36) areas, the rising rates of HR were markedly higher in the PC 4-skin group than in the "Ximen" (PC4), PC4-muscle and PC 4-periosteum groups (P < 0.01), and considerably higher in the "Housanli" (ST36), ST 36-skin, and ST 36-muscle groups than in ST 36-periosteum group (P < 0.05). The rising rates of MAP were significantly higher in the PC 4-skin and PC 4-muscle groups than in the "Ximen" (PC4) and PC4-periosteum groups (P < 0.01), and considerably higher in the ST36-skin group than in the "Housanli" (ST36), ST36-muscle and ST36-periosteum groups (P < 0.01), suggesting different effects of EA stimulation of different tissue layers in the same one acupoint. No significant differences were found between the "Ximen" (PC 4) and PC 4-muscle groups, among the "Housanli" (ST36), ST36-skin and ST 36-muscle groups in the rising rates of HR, between the "Ximen" (PC 4) and PC4-periosteum groups, and among the "Housanli" (ST36), ST 36-muscle and ST36-periosteum groups in the rising rates of MAP (P > 0.05). (3) The effect of the PC 4-skin group was significantly superior to that of the ST 36-skin group in increasing HR (P < 0.01), and the effect of the PC 4-muscle group was obviously stronger than that of the ST 36-muscle group in raising MAP (P < 0.01), suggesting different therapeutic effects of EA stimulation of the similar tissue in different meridian-acupoints. The effects of the "Ximen"(PC4) and "Housanli" (ST36) groups in raising both HR and MAP, and that of the PC 4-muscle and ST 36-muscle groups in upregulating HR, and that of the PC 4-periosteum and ST 36-periosteum groups in raising MAP were comparable (P > 0.05)., Conclusion: EA of different acupoints of the same one meridian, the similar structure of different meridian acupoints and different tissue layers of the same one acupoint have their own relatively specific effects in upregulating HR and MAP in hypotension plus bradycardia rats.

Objective: To observe the influence of electroacupuncture (EA) of different acupoints on changes of mean arterial pressure (MAP), heart rate (HR) and heart rate variability (HRV) in colorectal distension (CRD) rats, so as to analyze the specificity of actions of acupoints in relieving visceral pain and regulating activities of the autonomic nerve system., Methods: Forty-five Wistar rats were randomized into control, Zusanli (ST 36), non-acupoint, Neiguan (PC 6) and Taichong (LR 3) groups (n = 9/group). Under anesthesia, CRD was given to the rats by using an aerostat for 5 min. EA (2 Hz/15 Hz, 2 mA) was applied to bilateral ST 36, non-acupoint (1.0 cm lateral to ST 36), PC 6 and LR 3 for 15 min, respectively. Electrocardiogram of the cervico-chest lead was recorded by using a bioelectric amplifier, and MAP recorded by using a pressure transducer and an amplifier. Low frequency and high frequency of HRV were analyzed by Chart 5.0., Results: Following CRD, the HR, MAP, LF and LF/HF levels increased significantly in all the 5 groups (P < 0.05), while HF had no apparent changes (P > 0.05). Compared with the control group, 5 min and 15 min after EA ,and 10 min after ceasing EA, MAP values of ST 36 and PC 6 groups were decreased obviously (P < 0.05). The HR values of ST 36 and PC 6 groups at 15 min after EA and 10 min after ceasing EA were decreased markedly in comparison with those of each of the other groups during CRD (P < 0.05). The LF levels of both ST 36 and PC 6 groups at 5 min after EA, and those of ST 36, PC 6 and LR 3 at 15 min after EA were significantly lower than those of control group (P < 0.05). LF/HF levels of ST 36 and PC 6 at 10 min after ceasing EA were obviously lower than those of control group (P < 0.05). No significant differences were found among ST 36, non-acupoint, PC 6 and LR 3 groups in HR, LF and LF/HF (P > 0.05)., Conclusion: EA of ST 36 and PC 6 can suppress CRD-induced increase of MAP, HR and LF/HF, suggesting beneficial effects of EA in relieving visceral pain and mediating autonomic nerve system. The aforementioned effects of EA of LR 3 and non-acupoint are not obvious.

An amino acid sequence variant is defined as an unintended amino acid sequence change and contributes to product heterogeneity. Recombinant monoclonal antibodies (MAbs) are primarily expressed from Chinese Hamster Ovary (CHO) cells using stably transfected production cell lines. Selections and amplifications with reagents such as methotrexate (MTX) are often required to achieve high producing stable cell lines. Since MTX is often used to generate high producing cell lines, we investigated the genomic mutation rates of the hypoxanthine-guanine phosphoribosyltransferase (HGPRT or HPRT) gene using a 6-thioguanine (6-TG) assay under various concentrations of MTX selection in CHO cells. Our results show that the 6-TG resistance increased as the MTX concentration increased during stable cell line development. We also investigated low levels of sequence variants observed in two stable cell lines expressing different MAbs. Our data show that the replacement of serine at position 167 by arginine (S167R) in the light chain of antibody A (MAb-A) was due to a genomic nucleotide sequence change whereas the replacement of serine at position 63 by asparagine (S63N) in the heavy chain of antibody B (MAb-B) was likely due to translational misincorporation. This mistranslation is codon specific since S63N mistranslation is not detectable when the S63 AGC codon is changed to a TCC or TCT codon. Our results demonstrate that both a genomic nucleotide change and translational misincorporation can lead to low levels of sequence variants and mistranslation of serine to asparagine can be eliminated by substituting the TCC or TCT codon for the S63 AGC codon without impacting antibody productivity., (2010 Wiley Periodicals, Inc.)

Objective: To study the long-term outcomes and complications of giant cell tumors around the knee treated with en bloc resection and reconstruction with prosthesis., Methods: From January 1991 to March 2005, 19 patients (11 men, 8 women, average age 35.4 years) were treated in our hospital with en bloc resection and reconstruction with domestic prosthesis (15 hinge knee and 4 rotating-hinge knee). The distal femur was involved in 12 and the proximal tibia in 7 cases. Nine tumors were primary and 10 recurrent. All cases were Campanacci grade III. The affected limb functions were evaluated by the Musculoskeletal Tumor Society scoring system., Results: All patients underwent operation successfully with no complications. The mean follow-up time was 128.9 months (60 to 216 months). Apart from one patient who underwent amputation because of wound infection two years after reoperation, the range of knee motion of 18 patients was 30°-110°. The mean functional score of the affected limb was 22.7 (15 to 27 points). The length of the lower extremities was equal in nine cases; the affected limb was 2-9 cm shorter in the other ten cases. Prosthesis fracture and loosening developed in one, prosthesis aseptic loosening in three, and delayed deep infection and prosthesis loosening in two cases. The prosthesis loosening rate was 31.6%. One patient developed a proximal femur fracture., Conclusion: En bloc resection and reconstruction with prosthesis is a feasible method for treating giant cell tumor of bone around the knee. Complications related to the prosthesis, mainly prosthesis loosening and limb shortening, increase gradually with longer survival time., (© 2010 Tianjin Hospital and Blackwell Publishing Asia Pty Ltd.)

Objective: To investigate the functional specificity of acupoint by means of acupoint injection of Dobutamine Hydrochloride (DH)., Methods: Male SD rats were randomized into normal control (NC), EA-Neiguan (PC6) + acupoint injection (AI, EA-PC 6 + AI), EA-Zusanli (ST 36) + AI (EA-ST 36 + AI), and EA-intramuscular injection site (IMIS) + intramuscular injection (EA-IMIS+ IMI) groups,with 8 rats being in each group. EA (2 Hz/15 Hz, 3 mA) was applied to bilateral "Neiguan" (PC 6), "Zusanli "(ST 36) and muscular-injection site for 10 min. DH (a beta1-adrenoceptor agonist) solution (100 microg/kg) was injected intramuscularly into the gluteus maximus after termination of EA intervention. The left ventricular systolic pressure (LVSP), heart rate (HR) and + dp/dt max were recorded before and 2, 5, 15 and 30 min after injection of DH., Results: The values of LVSP, HR and + dp/dt max in EA-PC 6 + AI group were significantly higher than those in NC, EA-ST 36 + AI, and EA-IMIS + IMI groups (P < 0.01) 2 and 5 minutes after EA plus acupoint injection of DH. No significant differences were found between EA-ST 36 + AI and EA-IMIS+ IMI groups in LVSP, HR and +dp/dt max (P > 0.05). After injection of DH for 30 minutes, the values of LVSP, HR and + dp/dt max in EA-PC 6 + AI group were still significantly higher than those in NC group (P < 0.01)., Conclusion: EA-PC 6+ AI DH is significantly superior to EA-ST 36 + AI DH and EA-IMIS+ IMI DH in producing a stronger cardiac excitatory effect and a longer post-effect, showing a relative specificity of the acupoint in upregulating cardiac functional activities.

Specificity of acupoints is one of the basic theories of the acupuncture and moxibustion sciences and is a very important basis to guide the clinical treatment of acupuncture and moxibustion. However, the scientific foundation of specificity of acupoints is still unclear, which has greatly restricted to the clinical effect improvement of acupuncture and moxibustion and has influenced to academic status of acupuncture and moxibustion sciences both in China and abroad. In this paper, the recent related researches are reviewed and then the key effects and mechanism that the tissue structure of acupoints can decide and affect specificity of acupoints-organs effects are analyzed and explored. It is held that acupuncture at the different tissue structure of the different depth in the different or the same acupoints will cause changes of varying qualities or degrees of visceral function because of stimulation of the different receptors in the different tissues followed by excitation of varying afferent nerve fibers.

Acupuncture combined with drugs has been widely used in the treatment of various clinical conditions and can produce a stronger or more beneficial curative effect than either separate acupuncture or chemical drug does. However, the mechanisms underlying the additive or synergetic effects of acupuncture plus drugs are not well understood so far. Based on careful review and analysis of the literature accumulated, the authors proposed several aspects possibly involving the additive or synergetic effects produced by acupuncture plus drugs. For instance, acupuncture stimulation at acupoints may influence the drug concentration in blood and/or in target organ and the bioavailability of the drug via modulating the drug absorption, distribution and metabolism in the body. In addition, the curative effect of drug may also be strengthened by acupuncture stimulation via altering the signaling pathways of the targeted specific receptors and via potentiating the resultant responsiveness or sensitivity of the cells in the target organ in responding to the drugs administered. The ideas mentioned above may raise the novel clues for further studying the mechanisms underlying acupuncture-produced increase of the curative effects of medicines.

Deoxycytidine kinase (dCK) phosphorylates deoxycytidine, deoxyguanosine, and deoxyadenosine and plays an important role in the salvage pathway of nucleoside metabolism. dCK is also required for the phosphorylation of several antiviral and anticancer nucleoside drugs, with resistance to these agents often being associated with a loss or decrease in dCK activity. Data also indicate a role for dCK in immune function, and dCK inhibitors may provide treatment for immune disorders. To identify novel dCK inhibitors, the authors evaluated 2 existing biochemical assays, adapted both to high-throughput screening, and identified several series of hits. They also compared the potency of the hits between purified recombinant and endogenous enzyme. Meanwhile, they also developed a novel cell-based assay that rests on the rescue of cells from dCK-dependent cytotoxic agents such as AraC. A large number of compounds were tested using the 3 assays, and a strong correlation in potency was observed between the biochemical assay using endogenous enzyme and the cell-based assay. The hits identified in these screens have proved to be good starting points for the synthesis of much more potent tool compounds to further investigate the physiological functions of dCK and potentially lead to the development of therapeutic agents.

A series of potent piperidine-linked cytosine derivatives were prepared as inhibitors of deoxycytidine kinase (dCK). Compound 9h was discovered to be a potent inhibitor of dCK and shows a good combination of cellular potency and pharmacokinetic parameters. Compound 9h blocks the incorporation of radiolabeled cytosine into mouse T-cells in vitro, as well as in vivo in mice following a T-cell challenge.

A series of deoxycytidine kinase inhibitors was simultaneously optimized for potency and PK properties. A co-crystal structure then allowed merging this series with a high throughput screening hit to afford a highly potent, selective and orally bioavailable inhibitor, compound 10. This compound showed dose dependent inhibition of deoxycytidine kinase in vivo.