TA method for the content determination of methionine sulfoxide and methionine sulfone in compound amino acid injection (18AA-II) was established in order to investigate their level in 155 batches of this product, and to explore the reason for the generation of these two impurities.The determination was performed on an Agilent Poroshell 120 EC-C18 column with mobile phases of sodium acetate/tetrahydrofuran solution (A) and sodium acetate solution -acetonitrile-methanol (B, 200∶400∶400) (gradient elution) at the flow rate of 0.5 mL/min.The excitation wavelength and the emission wavelength of the fluorescence detector were 233 nm and 441 nm, respectively.The column temperature was 40 °C, and the injection volume was 8 μL.The contents of methionine sulfoxide and methionine sulfone from 155 batches of compound amino acid injection (18AA-II) was determined using this method, and the residual oxygen content was detected by headspace gas analyzer.The results showed that the linear range of methionine sulfoxide and methionine sulfone were 0.128 1-10.250 0 μg/mL (r = 0.999 9) and 0.261 0-10.440 0 μg/mL (r = 0.999 8), respectively.The limits of quantitation were 0.13 μg/mL and 0.26 μg/mL, respectively; the limits of detection were 0.04 μg/mL and 0.09 μg/mL, respectively.RSDs of precision, stability and repetitive test were all lower than 1.3%.The recoveries ranged 98.00%-100.79% (RSD = 1.15%, n = 9) and 98.19%-102.31% (RSD = 1.33%, n = 9).The content level of oxidized related substances from different manufacturers showed significant difference, showing relevance with the residual oxygen content to some extent, yet no significant correlation with the added amount of antioxygen (sodium pyrosulfite).The method is validated to be useful for the content control of methionine sulfoxide and methionine sulfone in compound amino acid injection (18AA-II).It is quite necessary to include the determination of oxidized related substance into the quality specification.Manufacturers should strengthen the control of remaining oxygen in their products.

Objective To identify the rale of NKX2-5 gene in cardiomyocyte differentiation and its mechanism.Methods P19 cells were divided into transfected and non-transfected groups.In the transfected group,P19 cells were with stable expression of NKX2-5 gene.The P19 cells were cultured in suspension for 4 days,and the formed aggregates were transferred to Petri dish for adherent culture.On days 4,8,12,and 16 of the adherent culture,the expressions of ct-saicomeric actin(α-SA)and cardiac troponin T(cTnT)were detected with double-labeling immunofluorescence and Western blot.The ultrastruetural changes were observed on day 16.Results In the transfected group,no expression of α-SA and cTnT was found on day 4,and the expression of these 2 proteins or co-expression existed on days 8,12,and 16.There were early cell junction and myofilament-like structure in the cytoplasm of some cells in the transfected group.In the non-transfected group,these 2 proteins were negative,and no differentiated cell was found.Conclusion Stable expression of NKX2-5 gene can induce cardiomyocyte differentiation from P19 cells,but the P19 cells with stable expression of JVKX2-5 gene is not suitable to be an in vitro model of cardiac development.

Field operations,work and war on plateau is an important part of future war. It is necessary to study the early treatment and evacuation of scull injury. Research and survey were made in 7 field hospitals and hundreds of records were summaried before treatment criterion in early phase was put forward for patients on plateau as well as the evacuation in different level and stage,which aims to actively save patients' lives within limited time and reduce the death rate and prevent deformity. References are provided for war,training,and construction in battlefield.

Objective To investigate a method for the purification of the N?terminal peptide fragment(NT)of the myocardial calcium channel Cav1.2,and characterize its interaction with calmodulin(CaM). Methods EscherichiacoliBL?21 cells were transformed with plasmid pGEX?6p?3/NT harboring the NT?GST fusion gene. The cells harboring pGEX?6p?3/NT were cultured and protein expression was induced with isopropyl?β?D?thiogalactoside(IPTG). Then,the GST?NT fusion protein was purified by using glutathione Sepharose 4B(GS?4B)beads. GST was cleaved off with the PreScission protease,and SDS?PAGE was performed to detect the purity and relative molecular weight of the purified peptide. Further, GST pull?down assay was performed to characterize the interaction of the NT peptide with CaM. Results SDS?PAGE analysis showed that the NT peptide was successfully purified,with high purity. Results of the GST pull?down assay showed that the NT peptide could interact with CaM. Conclusion This study establishes a method for the purification of the NT peptide and lays the foundation for further research on the interaction partners and biological functions of NT.

Objective To evaluate the clinical efficacy and safety of stereotactic radiotherapy (SRT) combined with bevacizumab for brain metastases in patients with lung adenocarcinoma.MethodsThe clinical data of 95 patients with brain metastases of lung adenocarcinoma were retrospectively analyzed, including 36 patients treated with SRT and bevacizumab (bevacizumab group) and 59 patients treated with SRT, corticosteroids, and mannitol (traditional drug group).The tumor response rate, peritumoral edema control rate, improvement in Karnofsky Performance Scale (KPS) score, and adverse reactions were analyzed.Results Compared with the traditional drug group, the bevacizumab group had significantly higher tumor response rate (P=0.033) and peritumoral edema control rate (P=0.000) at 1-4 weeks after treatment, as well as an obvious improvement in KPS score and reduced doses of corticosteroids and mannitol.In addition, the adverse reactions in the bevacizumab group were mild and controllable.Conclusions SRT combined with bevacizumab for brain metastases in patients with lung adenocarcinoma can achieve higher short-term tumor response rate and peritumoral edema control rate and improve patients' quality of life.

Objective To discuss the clinical effect of the ramified musculocutaneous flap pedicled with the descending branch of lateral circumflex femoral artery treatment of bones and arthrsis of extremity in- fections with soft tissue defects.Methods The muscle flap blooded with the muscular branch of lateral fem- oral muscle and musculocutaneous flap blooded with the musculocutaneous branch were designed,all of which were pedicled with the descending branch of lateral circumflex femoral arteryIn clinic24 cases of bones and arthrosis of extremity infections with soft tissue defects were treated with this kind of ramified musculocutaneous flap.Results Of the 24 cases23 cases were survived while 1 case was lost16 cases were healed at stageⅠ8 cases were healed at stageⅡSinus has formated in 3 casesone of which twicebut they were healed in one year with the treatment of debridmentsFour cases with osteomvelitis and bone defect were treated with bone grafting in the later 6～8 months after the wound has healedTwenty-two cases were followed-up for 6～20 monthsinfcetiou didn't recur.Conclusion This kind of ramified musculocutaneous flap has such ad- vantages as longer blood vessel pediclefilling the defects completely flexible application and stronger anti-in- fectionthat it may be an effective way in treating bones and arthrosises of extremity infections with soft tissue defects.

Acetabulum ; injuries ; Adult ; Bone Plates ; Dermatologic Surgical Procedures ; Female ; Follow-Up Studies ; Fractures, Bone ; diagnostic imaging ; physiopathology ; surgery ; Humans ; Male ; Middle Aged ; Tomography, X-Ray Computed ; Traction ; Young Adult

A Kinase Anchor Proteins ; metabolism ; Animals ; Cell Cycle Proteins ; metabolism ; Lipopolysaccharides ; pharmacology ; Liver ; metabolism ; pathology ; Protein Kinase C ; metabolism ; Rats ; Rats, Sprague-Dawley ; Signal Transduction

Objective To observe the brain activation of interictal epiletiform discharges(IEDs) and to localize the epileptogenic foci of epilepsy.Methods The electroencephalogram(EEG)and functional MRI data of 12 focal epileptic patients were acquired using a combination of EEG and functional MRI simultaneously.The IEDs onset time detected with EEG were set as the time parameters in an event- related paradigm of functional MRI analysis.The spatial and temporal characters of IEDs activation were analyzed in detail.In order to confirm the consistency of this method,all patients were scanned repeatedly and the results were correlated with clinical evaluation.Results Of the 12 patients,valid data from EEG- fMRI were obtained from 10 patients in a total of 18 sessions..Compared with the structural foci,the epileptic foci localization results of eleven sessions were good,five sessions were fairly good,and two sessions were poor.The results obtained from six patients in two separate sessions were concordant.respectively.Moreover,thalamic activation was detected in ten sessions,cerebellar activation was detected in all sessions,and the deactivation was found in the default mode loci in nine sessions. Conclusion The method of performing EEG and fMRI simultaneously can potentially be a useful tool in epilepsy research.

BACKGROUND: Local soft tissue lysis has been widely used in clinical rehabilitation treatment of chronic soft tissue injuries, and has achieved certain clinical outcomes. However, it is an invasive treatment, and there is little information about the pathological changes and prognosis of the local tissues post-treatment. OBJECTIVE: To observe the repair process and pathologic changes of the muscles after local soft tissue lysis with several needles. METHOD:Songjin-needle,Yinzhi-needle,Changyuan-needle,Xiaozhen-needle,Ren-needle and Pi-needle were inserted into the belly of rat gastrocnemius gastrocnemius with a depth of 0.5-1.0 cm, twice or thrice. Then the rats were killed at 3, 7, 14, 21 and 30 days after acupuncture. The corresponding parts of tissues were removed and fixed with paraformaldehyde, and stained with hematoxylin and eosin and immunohistochemistry to observe the histomorphological features and pathological changes under light microscope. The CellSens-Entry 1.15 software was used for image acquisition, and the number of fibroblast cells and microvessel density were calculated. The integral absorbance values of collagen type Ⅰ were calculated by Image Pro plus 6 software. RESULTS AND CONCLUSION: On day 3 after intervention, muscle fiber dissolution, necrosis and inflammatory cell infiltration were visible. On day 7, there was a reduction in inflammatory cells, fibroblast activation and proliferation, and granulation tissue hyperplasia. On day 14, there was local accumulation of collagen with substantial granulation tissues. On day 21, collagen fibers decreased and granulation tissues organized. On day 30, there was local scar formation. Immunohistochemical staining results showed that there was collagen type Ⅰ in the surgical area after intervention. The microvessel density, fibroblast count and integral absorbance value increased firstly and then decreased, which showed significant differences among groups (P < 0.01). These findings imply that the new types of needles for local soft tissue lysis do certain damage to the gastrocnemius muscle, and the inflammatory changes of the local tissue and granulation tissue organized will result in local scar formation after loosening therapies.