Objective To establish a rapid test method for the determination of five sweeteners and two preservatives in the milk beverage by HPLC.Methods Trough a set of pretreatment such as dilution,protein precipitation,high-speed centrifugation,the samples were tested.With NUCLEODUR C18 RP-column separation,phosphate buffer(pH=5.0) and acetonitrile were used as the mobile phase,gradient elution analysis was conducted,the detective wavelength was 205nm.Results The separation of one sample was achieved within 15 min,five sweeteners and two preservatives could be well separated,the linear range was 5.00-100.0 mg/L,r≥0.9993,RSD was 1.5%-4.2%(n=6),the average recovery rate was 85.2%-108%.Conclusion This method is simple and quick with better reproducibility and selectivity,it is an effective way to determine sweeteners and preservatives in milk beverage.

Objective To evaluate application value of plasma tumor type M2 pyruvate kinase (TU M2-PK) in the treatment effect monitoring in breast cancer. Methods TU M2-PK was determined by ELISA in breast cancer patients (n = 63 ), benign breast disease patients (n = 22 ) and health controls (n = 40).The receiver operation characteristic (ROC) analysis was performed as compared with CA15-3 and CEA. Results ROC analysis showed the cut-off was set at 14. 1 U/ml for TU M2-PK ( sensitivity 46. 0% ;specificity 86. 0% ), and the diagnosis efficacy of TU M2-PK was higher than CA15-3 and CEA. The level of TU M2-PK was significantly higher in breast cancer patients (13. 3 U/ml) than that in health controls (7. 2 U/ml, U = 408. 5, P < 0. 05 ) and in benign breast disease patients ( 11.1 U/ml, U = 509.0,P < 0. 05 ). With the progression of breast carcinoma, the level of TU M2-PK as well as the positivity was increased. TU M2-PK concentration was higher in patients with lymph node metastasis (23. 3 U/ml ) than those without metastasis ( 10. 9 U/ml, U = 237. 0, P < 0. 01 ). The level of TU M2-PK correlated with therapy response. An elevated level of TU M2-PK was found preclinically in recurrent disease patients, and the levels decreased in the patients, which showed sensitive to chemotherapy. The TU M2-PK level was kept at baseline in patients with stable disease. Conclusion TU M2-PK is helpful in the diagnosis of breast cancer, and it is a valuable tumor marker for disease monitoring, therapy control and prognosis evaluation in breast cancer.

OBJECTIVE To analyze the application of antibacterials in liver transplanted patients of our hospital in order to improve the rational use of antibacterials in perioperative period of liver transplantation.METHODS According to the criteria of DDD and DUI recommended by WHO,a retrospective study of the application of antibacterials in 82 patients with liver transplantation who discharged hospital during from Jun 2003 to Jun 2005 was made.RESULTS Among the 82 patients,100.0% patients had received antibacterials and 48 patients received combined medication,in which 45.10% used two kinds and 13.41% used three kinds.The longest medication time was 49 days while the shortest was 10 days.Nineteen antibacterials′ DUI were all less than one except meropenem whose DUI was 1.03.CONCLUSIONS This study proved that patients with liver transplantation in our hospital received rational antibacterials.

[Summary] Limb numbness is a common symptom of diabetic peripheral neuropathy(DPN),which is similar to lumbar vertebra disease and spinal cord tumor compression syndrome. Diabetes mellitus (DM) combined with lumbar spondylosis and spinal cord tumor compression is rarely occurred and is difficult to distinguish from DPN . Here we reported a case of DM combined with spinal neurilemoma that was misdiagnosed as DPN for half a year.

Objective To investigate the impact of angiotension II (Ang II ) on the activation of nuclear factor-kappaB (NF-?B) signaling transduction pathway in cultured rat pancreatic stellate cells (PSCs). Methods Growth-arrested rat PSCs were incuhated for indicted time intervals with increasing concentrations of Ang II . The DNA binding activity of NF-?B was determined using electrophoretic mobility shift assay (EMSA). Western blot analysis was used to examine the degradation of inhibitory proteins of NF-?B (I?Bs). Expression of monocyte chemoattractant protein-1 (MCP-1) mRNA and protein were assessed by Northern blot and enzyme-linked immunosorbent assay (ELISA), respectively. Results Treatment of cells with Ang II led to a biphasic activation of NF-?B, which peaked at 15 min and 6 h later, and it was correlated with differential degradation of I?B? and I?B?. Ang II -induced NF-?B activation was greatly inhibited by antioxidants pyrrolidine dithiocarbamate ( PDTC), diphenylene iodonium. and N-acetylcysteine, suggesting the involvement of oxidative stress in this process. In PSCs, Ang II induced a dose-dependent increase in the expression of MCP-1 and this effect was markedly inhibited by blocking NF-?B activation with MG132 and PDTC, indicating that Ang II -induced MCP-1 gene expression was NF-?B-dependent. Conclusion The present results suggest that Ang II , through an oxidative stress-dependent mechanism, may activate a functional NF-?B signaling pathway in PSCs, through which it may participate in pancreatic inflammation and fibrosis.

Objective To investigate the clinical characteristics and surgical treatment for primary presacral tumors.Methods The clinical data of 42 patients of primary presacral tumors from January 2013 to May 2015 were analysed retrospectively.Results Of the 42 patients,16 cases were asymptomatic while 26 patients had discomfort at the sacral or abdominal region,or difficulty in urinating or defecation.90％ of the cases were digital rectum examination (DRE) positive.Among the 42 patients 36 cases underwent surgical treatment,1 case underwent radiotherapy,5 cases refused surgical treatment.Among those receiving surgical resection,28 cases had trans-abdominal surgery and 4 cases had trans-sacral surgery,while 3 cases had trans-abdominal & trans-sacral surgery,1 case had trans-abdominal and perineal surgery.Tumors were completely resected in 31 cases,and palliatively resected in 5 cases.3 cases suffered from intra-operative presacral hemorrhage.1 case with delayed hemorrhage required surgical intervention.2 cases from incision infection recovered after wound disinfection and dressing.3 cases had postoperative hip or leg numbness;1 case with high fever was cured by intensive antibiotics treatment.Conclusion The low incidence of presacral tumors makes early detection difficult.A diagnosis can be obtained by a positive DRE combined with CT or MRI results.Resection is a therapy of choice after biopsies.

Corynebacterium glutamicum SA001 is a mutant with lactate dehydrogenase (ldhA) deletion. In order to increase metabolic flux from isocitrate to succinate, and to improve the production of succinate under anaerobic conditions,we transducted the gene aceA coding isocitrate lyase (ICL) from Escherichia coli K12 into Corynebacterium glutamicum SA001 (SA001/pXMJ19-aceA). After 12 h aerobic induction by adding 0.8 mmol/L of IPTG, the recombinant strain was transferred to anaerobic fermentation for 16 h. Succinate reached 14.84 g/L, with a productivity of 0.83 g/(L x h). Compared to C. glutamicum SA001, the activity of ICL of the recombinant strain was increased 5.8-fold, and the succinate productivity was increased 48%. Overexpression of isocitrate lyase will increase the metabolic flux of glyoxylate bypass flowing to succinate.
Corynebacterium glutamicum ; genetics ; metabolism ; Escherichia coli ; enzymology ; genetics ; Gene Deletion ; Industrial Microbiology ; Isocitrate Lyase ; biosynthesis ; genetics ; L-Lactate Dehydrogenase ; genetics ; Succinic Acid ; metabolism ; Transduction, Genetic

ObjectiveTo evaluate the feasibility of CT coronary angiography (CTCA) in patients with arrhythmia using 320-detector row CT.MethodsThirty-one patients with persistent atrial fibrillation and 8 patients with premature ventricular contraction were enrolled in this study.All patients underwent 320-detector row CTCA.CT image quality was evaluated with 4-point grading scale by two radiologists.Interobserver agreement was evaluated by Kappa statistics.The radiation dose was calculated.ResultsIn total510 coronary segments,496 (97.2％) segments met diagnostic standard.The mean effective dose was (12.7 ± 4.8) mSv in this study.There was a good agreement in image quality scoring between the two reviewers (Kappa =0.72 ).Conclusion 320-detector row CTCA is feasible in patients with atrial fibrillation and premature ventricular contraction.Arrhythmia may not be considered as a contraindication to CTCA.

BACKGROUND: The study of isolation, purification, culture, cell labeling, inducing factors, effects of gene transfection on cytobiology, cell carrier construction, and time window for back transplantation of cell compound pertaining to marrow stromal cells (MSCs) is still in its infancy. OBJECTIVE: To search for an in vitro culture method that can be simply and effectively obtained. DESIGN, TIME AND SETTING: The present cytological in vitro experiment was performed at the Beijing Institute of Genome, Chinese Academy of Sciences between June 2006 and July 2007. MATERIALS: Eight specific pathogen-free New Zealand rabbits, aged 6 weeks, were provided by the Laboratory Animal Center, Institute of Genetics and Development, Chinese Academy of Sciences. METHODS: Under sterile condition, 1 mL rabbit bone marrow was taken and diluted with D-Hanks solution. Following centrifugation and subsequent supernatant removal, bone marrow was re-suspended using dulbecco's modified eagle's medium (DMEM) for single cell suspension. Next, single cell suspension was dropped onto the liquid surface of equal-volume lymphocyte separation medium (density: 1.077). Subsequent to centrifugation, cloudlike mononuclear cell layer was collected and re-suspended with DMEM containing 20% fetal bovine serum. The cells were inoculated at lxl0/cm2 and purified by adherent method. When 70%-80% of flask bottom was covered, cell digestion and passage was performed.MAIN OUTCOME MEASURES: Cell growth was observed with an operating microscope. Surviving cells were counted by Trypan blue viability test. Cell identification was performed by hematoxylin-eosin staining. Through the use of 3-(4, 5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide assay, cell viability was detected to observe the cellular resuscitation of the cultured cells following cryopreservation. RESULTS: Twenty-four hours after inoculation, cells began to adhere to the wall, exhibiting short shuttle- or triangle-shaped appearance with different sizes of cellular processes. Three days later, adherent cells began to divide, and cell clusters could be found in some areas; One week later, most of cells exhibited scattered fibroblast-like growth; After passage, cells were evenly distributed with long shuttle-shaped appearance and arranged orderly. Following 3 passages, there wound be 5×106-1×107 adherent cells in 1 mL MSC suspension. Approximately 90% of MSCs survived and identified as mononuclear cells. Cells vigorously grew at days 1-6 after inoculation and reached a peak level at day 8, followed by a viability decline. After 56 days of resuscitation, frozen cells displayed a good and rapid growth. CONCLUSION: Highly purified MSCs can be acquired by gradient centrifugation of lymphocyte separation medium. Enough seeded cells can be obtained by in vitro culture and the cellular viability does not change after frozen preservation and resuscitation.

BACKGROUND: SOX-9 plays an important role in occurrence and development of cartilaginous tissues,enhances agglutination of mesenchymal cells,has structural domains of transcriptional activation,and can directly regulate the transcription,OBJECTIVE: To construct pDC316-SOX-9 plasmid for transfection of rabbit bone marrow stromal cells (BMSCs) using SOX-9 gene,and to investigate the effects of SOX-9 gene on growth characteristics of BMSCs and product expression. DESIGN,TIME AND SETTING: The cell gene engineering in vitro experiment was performe,d at the Beijing Institute of Genome, Chinese Academy of Scienees from June 2006 to January 2007.MATERIALS: Eight SPF New Zealand rabbits aged 6 weeks were used for culture of BMSCs.METHODS: pDC316-SOX-9 plasmid was used for transfection of rabbit BMSCs by liposome mediated method,Cell transfection included a SOX-9 plasmid transfection group,a blank plasmid group and a blank control group (only treatment of liposome). MAIN OUTCOME MEASURES: Cell morphology; growth activity; The SOX-9 protein expression in rabbit BMSCs were detected by immunohistochemistry,hematoxylin-eosin staining,reserve transcriptase-polymcrase chain reaction (RT-PCR) and enzyme-labeled immunosorbent assay (ELISA). RESULTS: Some cell colonies were detected at 4 days after pDC316-SOX-9 plasmid transfection.Spindle-shaped cells were collected after clone amplification.Cells in the blank control group gradually died over time.Cell activities in the SOX-9 plasmid transfeetion group and the blank plasmid group significantly prolonged,reached a peak at 2 weeks of transfection,and then gradually decreased.At 6 days,BMSCs were yellow in the SOX-9 plasmid transfection group following immunohistochemistry,expressing SOX-9 protein.Hematoxyliu-eosin staining showed many dikaryocytes,rich cell proliferation,similar to chondroblasts.No SOX-9 protein expression and unproductive cell proliferation in the blank plasmid group.SOX-9 mRNA was detected in the SOX-9 plasmid transfection group,but SOX-9 mRNA was not detected in the blank plasmid group and blank control group.SOX-9 levels were significantly higher in the SOX-9 plasmid transfection group than in the blank plasmid group and blank control group at 24,48 and 72 hours,1,2 weeks (P＜ 0.01).CONCLUSION: Rabbit BMSCs were successfully transfected with pDC316-SOX-9 plasmid to enhance cell growth activity and to persistently stably secrete SOX-9 protein,resulting in the differentiation of BMSCs into cartilages.