A simple and effective method for the duplex PCR detection was developed by using sequences of exogenous gene(RDV MP-)and endogenous gene(Zein)as templates for PCR amplification.The results of routine PCR amplification for RDV MP-gene in transgenic maize suggested that RDV MP-gene can stably inheritate in transgenic plants and their progenies;The duplex PCR detection of all negative and part positive samples that obtained by routine PCR amplification confirmed that above negative results were exact,also showed that the quality of extracted DNA can meet the need of PCR amplification.The error ratio of negative samples was 1.4%.The method used in this study was simple and credible and can be used to detect transgenic plants and their products.

Aged ; Antineoplastic Combined Chemotherapy Protocols ; therapeutic use ; Bleomycin ; administration & dosage ; analogs & derivatives ; Carcinoma, Squamous Cell ; drug therapy ; Cisplatin ; administration & dosage ; Combined Modality Therapy ; Drugs, Chinese Herbal ; therapeutic use ; Esophageal Neoplasms ; drug therapy ; Female ; Humans ; Male ; Middle Aged ; Mitomycin ; administration & dosage ; Phytotherapy

A rapid, sensitive and convenient LC-MS/MS method was developed for the determination of γ-aminobutyric acid (GABA) in human plasma. d2-γ-Aminobutyric acid (d2-GABA) was synthesized as internal standard (IS). After extraction from human plasma by protein precipitation with acetonitrile, all analytes were separated on a Luna HILIC column (100 mm x 3.0 mm, 3 μm) using an isocratic mobile phase of water: acetonitrile: formic acid (20 : 80 : 0.12) with a flow rate of 0.5 mL x min(-1). Acquisition of mass spectrometric data was performed in multiple reaction monitoring mode (MRM) in positive electrospray ionization using the transitions of m/z 104 --> 69 for GABA and m/z 106 --> 71 for d2-GABA. The method was linear in the concentration range of 5.00 to 1 000 ng x mL(-1). The intra- and inter-day precisions were within 9.9%, and accuracy ranged from 99.1% to 104%, within the acceptable limit across all concentrations. The method was successfully applied to a pharmacokinetic study of GABA tablets in healthy Chinese volunteers.
Chromatography, Liquid ; Humans ; Tandem Mass Spectrometry ; gamma-Aminobutyric Acid ; blood

Objective To express the plasminogen activator(Pla) of Yersinia pestis and one of its gene fragments,and to detect their immunological reactivity.Methods The pla gene and its specific gene fragment pla-c were amplified by PCR using the EV76 strain as a template.PCR products were then ligated with the plasmid pET32a (+).The recombinant plasmids pET32a (+)-pla and pET32a (+)-pla-c were subsequently trausformed into E.coli BL21 (DE3).The expressed products were purified by HIS affinity chromatography,and their immunological reactivity was detected by Western blotting.Results The recombinant Pla(52.8 × 103) was expressed as inclusion bodies,and the recombinant Pla-c protein (24.0 × 103) was expressed in the soluble form.These two recombinant proteins reacted with anti-Yersinia pestis EV76 rabbit sera.Conclusions The recombinant Pla and its specific fragments have displayed immunological reactivity,and can be served as an alternative diagnosis method for Yersinia pestis.

Acupuncture Therapy ; Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Periarthritis ; physiopathology ; therapy ; Shoulder Joint ; injuries ; physiopathology ; Young Adult

Adolescent ; Adult ; Female ; Humans ; Male ; Meridians ; Middle Aged ; Moxibustion ; Spondylitis, Ankylosing ; therapy ; Young Adult

Objective to investigate the value of diffusion-weighted echo-planar MR imaging in the diagnosis of head and neck lesions.Methods Fifty-seven patients with 85 head and neck lesions were enrolled in the study,including 22 patients with 22 malignant tumors,13 patients with 13 benign tumors, 13 patients with 17 cystic and liquefactive lesions(including 8 patients with 12 cystic lesions,4 patients with 4 tumor necrosis,1 patients with 1 abcess)and 33 lymph nodes.The lesions were all confirmed by operation and clinical follow up.Echo-planar difffusion-weighted imaging (DWI)was performed with different b values (0,500,and 1,000s?mm~(-2)),and the apparent diffusion coefficients (ADCs)were measured.Results Malignant and benign tumors had different characteristics in DWI with different b values.With the increase of b value,the signal intensity of tumor/spinal cord ratio decreased quickly in DWI in benign tumors,while the signal intensity of tumor/spinal cord ratio remained similar in DWI in malignant tumors.The mean ADC value of'malignant tumors[(0.78?0.24)?10~(-3)mm~2? s~(-1)] was significantly lower than that of benign tumors [(1.48?+0.20)?10~(-3)mm~2?s~(-1)] (t = 8.9,P

Objective Renal ischemia-reperfusion may cause myocardial injury and dexmedetomidine ( DEX)is a new alpha-2 adrenergic agonist which has the effects of antisympathia , sedation and analgesia.The article was to observe the effects of different doses of dexmedetomidine on myocardial injury induced by renal ischemia -reperfusion(I/R) in Rats. Methods 40 male Wistar rats were randomized into 5 groups(n=8 each)using a random number table: sham operation group(isolation of bilateral renal pedicles without ligation), I/R group (3 hours′reperfusion 120 minutes after the right nephrectomy and the ligation of left renal artery ), DEX low dose group, DEX middle dose group and DEX high dose group (DEX 25, 50, 100 μg/kg were respectively injected intraperitone-ally in rats of the three groups plus 3 hours′reperfusion after 120 minutes′ischemia ) .Blood samples were collected at 3 hours′reper-fusion to determine serum creatinine(Cr) and blood urea nitrogen(BUN) concentrations.Kidney and myocardial specimens were ex-tracted for microscopic examination and IL-10,TNF-αcontent were detected by ELISA. Results In sham operation group, renal structure was normal.In I/R group, a great amount of erythrocyte infiltration and interstitial infiltrating inflammatory cells were found in glomerulus and a lot of exfoliative cells were found in renal tubules .In DEX low dose group , erythrocyte infiltration and interstitial infiltrating inflammatory cells were found in glomerulus and a few exfoliative cells were found in renal tubules .In DEX middle dose group, erythrocyte infiltration and interstitial infiltrating inflammatory cells were found in partial glomerulus and a few exfoliative cells were found in renal tubules .In DEX high dose group , erythrocyte infiltration and interstitial infiltrating inflammatory cells in partial glomerulus were found and rare exfoliative cells were found in renal tubules .In sham operation group , cardiomyocytes were arranged in perfect order and normal structure , and chromatins and cytoplasms were in uniform distribution .In I/R group, edema and spongiform were obvious in cardiomyocytes , and focal coagulative necrosis was observed along with a great amount of infiltrating inflammatory cells and rare flaky bleeding .In DEX low dose group , edema and spongiform were found in cardiomyocytes , and focal coagulative necrosis was observed along with a great amount of infiltrating inflammatory cells and rare flaky bleeding .In DEX middle dose group , edema was found in cardiomyocytes , and mini focal coagulative necrosis was observed along with a small amount of infiltrating inflammatory cells.In DEX high dose group , edema was found in cardiomyocytes along with a small amount of infiltrating inflammatory cells .Com-pared with sham operation group , Cr, BUN concentrations in serum and IL-10,TNF-αcontents in kidney tissue and myocardium signif-icantly increased in I/R group, low dose group, middle dose group and high dose group(P<0.05).Compared with I/R group, IL-10 contents kidney tissue and myocardium significantly increased in small dose group (P<0.05).Cr ([167.11 ±18.81, 135.46 ± 9.80, 114.29 ±12.50, 100.15 ±8.81]μmol/L), BUN ([13.42 ±1.25, 11.73 ±1.15, 10.27 ±0.78, 9.28 ±0.52] mmol/L) concentrations in serum and TNF-αcontents in kidney tissue ([578.45 ±30.59, 530.76 ±20.59, 482.23 ±27.12, 423.14 ± 21.16]ng/L) and myocardium ([565.00 ±37.66, 517.82 ±36.89, 469.99 ±32.43, 407.41 ±23.77] ng/L) significantly de-creased in a dose-dependent manner in low , middle and high groups (P<0.05).Microscopic examination showed that the pathological changes of both kidney tissue and myocardium were significantly attenuated in low , middle and high dose group . Conclusion Dexmedetomidine can allenuate myocardial tissue injury induced by renal ischemia -reperfusion in a dose-dependent manner in rats and its mechanism may be involved with the inhibition of inflammatory factors .

BACKGROUND:In vitro experiments have demonstrated that the biodegradable mesh-like microporous baloon made of macromolecular materials has obvious advantage of anti-leakage, which is capable of maintaining calcium homeostasis, has no inhibitory effects on cel growth and on microscopic interdigitation formation between new bone and bone cement. OBJECTIVE:To evaluate the therapeutic effects of biodegradable mesh-like microporous baloon with calcium bone cement on vertebral fractures based on animal experiments. METHODS:The fracture model was established in 48 New Zealand rabbits, in which a bone dril was introduced after successful puncture at sites near left low extremity of the femur. These rabbit models were randomized into two groups: experimental group with calcium phosphate bone cement and biodegradable mesh-like microporous baloon and control group only with calcium phosphate bone cement. Clinical parameters such as blood cel count, biochemistry, and CT/X ray were examined at 1, 3 and 6 months after implantation of the baloon and bone cement. After that, the specimens were fixed for pathological analysis. RESULTS AND CONCLUSION:The operation was performed under general anesthesia with no eventful infusion of bone cement. The expansion of baloon was satisfactory without definite extravasation of bone cement in the experimental group. In the control group, cement diffusion was found with pulmonary embolism occurring in three New Zealand rabbits. No statistical significance for blood cel counts and biochemistry was found between pre- and postoperation or between two groups. The materials in the two groups had favorable biocompatibility with injured bones without obvious immunological response. In the experimental group, the baloon wal was thinned and partial bone tissues grew into the cement at 1 month; at 3 months, a large amount of bone tissues grew into the cement and cement volume diminished; at 6 months, the baloon disappeared and only a smal amount of cement left in the bone tissues. In the control group, it was difficult to determine when the cement degraded. The biodegradable mesh-like microporous baloon combined with calcium bone cement is superior to bone cement alone in the management of vertebral fractures.

Objective To investigate the safety and effectiveness of Solitaire FR stent mechanical thrombectomy combined with 5F Navien catheter aspiration technique for the treatment of acute middle cerebral artery occlusion.Methods From February 2016 to May 2016,the case data of 11 consecutive patients with acute middle cerebral artery occlusion treated with Solitaire FR stent mechanical thrombectomy combined with 5F Navien catheter aspiration technique at the Department of Neurosurgery,Xuanwu Hospital,Capital Medical University were enrolled retrospectively.The age,gender,National Institutes of Health Stroke Scale (NIHSS) score on admission,thrombolysis in cerebral infarction (TICI) grade,operation-related complications,NIHSS score at 24 h after operation,and modified Rankin Scale (mRS) score at day 90 after operation were documented.The relevant data differences before and after treatment were analyzed with the SPSS software.Results Among the 11 patients with acute middle cerebral artery occlusion,7 were men and 4 were women,the age ranged from 44 to 78 years,and the mean age was 66 ± 11 years.The preoperative and postoperative NIHSS scores were 18 ± 3 and 8 ±4 respectively.There was significant difference before and after treatment (t =5.327,P ＜ 0.01).The recanalization of occluded middle cerebral artery achieved successfully.The follow-up at day 90 after operation,the good prognosis of the patients (mRS 0-2) was in 7 cases.No complications associated with severe M1 segment catheter operation occurred.Conclusion The preliminary experience showed,Solitaire FR stent mechanical thrombectomy combined with 5F Navien catheter aspiration technique for the treatment of acute middle cerebral artery occlusion was a safe and effective endovascular mechanical thrombectomy method.