Objective To evaluate the efficacy of recombinant human interferon α-2b gel onflooding prevention in loop electrosurgical excision procedure (LEEP) during and after the operation.Method The 152 patients with cervical disease were randomly divided into three groups: group A(recombinant human interferon α-2b gel treatment group), group B (hypophysin treatment group)and groupC(control group). Results The amount of intraoperative bleeding and the amount of post-operative bleedingof the group A was (2.57±1.19) ml and (2.24±1.75) ml, the group B was (3.00±1.49) ml and (45.86±26.14) hal, the group C was (45.62±39.57) ml and (56.90±41.90) ml, respectively. There was no differencebetween group A and group B in the amount of intraoperative bleeding (P>0.05), but significantly less thangroup C (P<0.01). There was no difference between group B and group C in the amount of post-operativebleeding (P>0.05), but significantly much more than group A (P<0.01). Conclusion Recombinanthuman interferon α-2b gel is effective on reducing intraoperative and post-operative bleeding in LEEP.

Guided by the antioxidant activity, the EtOAc-soluble and n-butanol-soluble fractions of the 50% methanol extract of Lonicera japonica leaves were isolated and purified by various chromatorgraphic methods, and the structures were identified by spectral analysis and comparison to the data reported in literature. As a result, nine compounds were obtained and identified as 5-O-caffeoylquinicacid (1), chlorogenicacid (2), 4-O-caffeoylquinicacid (3), luteolin-7-O-[α-L-arabinopyranosyl-(1 --> 6)] -β-D-glucopyranoside (4), luteoloside (5), 3,4-di-O-caffeoylquinic acid (6), 3,5-di-O-caffeoylquinic acid (7), 4, 5-di-O-caffeoylquinic acid (8) and luteolin (9). The antioxidant activity of the nine compounds were determined by using DPPH free radical scavenging method, and ascorbic acid was used as a positive control. Their antioxidant activities from high to low were 5 > 9 > 2 > 8 > 7 > 6 > 1 > 3 > 4. Among them, luteoloside (5) had the strongest antioxidant activity with an IC50 of 0.018 18 g x L(-1), and luteolin (IC50 0.023 6 g x L(-1)) and chlorogenicacid (IC50 0.035 17 g x L(-1)) ranks No. 2 and 3. Furthermore, the antioxidant activity of luteoloside and luteolin were stronger than that of ascorbic acid (IC50 0.027 54 g x L(-1)). These results gave a basis for the further study and utilization of L. japonica leaves.
Antioxidants ; chemistry ; isolation & purification ; Drugs, Chinese Herbal ; chemistry ; isolation & purification ; Lonicera ; chemistry ; Mass Spectrometry ; Molecular Structure ; Plant Leaves ; chemistry

AAV-ITR gene expression mini vector is a double-strand or single-strand DNA that only contains inverted terminal repeats of adeno-associated virus, cis-elements and gene of interest and does not contain any other foreign DNA sequences. We prepared Bac-ITR-EGFP and Bac-inrep. Spodoptera frugiperda cells were infected with Bac-ITR-EGFP (P3) and Bac-inrep (P3). Up to 100 μg of AAV-ITR-EGFP gene expression mini vectors were extracted from 2 x 10(7) cells of Sf9 72 h after infection. The gel electrophoresis analysis shows that most forms of AAV-ITR-EGFP gene expression mini vector were monomer and dimer. The mini vector expression efficacy was examined in vitro with HEK 293T cells. The EGFP expression was observed at 24 h after transfection, and the positive ratio reached 65% at 48 h after transfection.
Animals ; Baculoviridae ; DNA, Single-Stranded ; Dependovirus ; Gene Expression ; Genetic Vectors ; HEK293 Cells ; Humans ; Sf9 Cells ; Terminal Repeat Sequences ; Transfection

Methods of ELISA, nonradioactive molecular hybridiz ation and RT-PCR were applied in the detection of rice grassy stunt virus (RGSV ). The detection sensitivity of indirect ELISA using antiserum against fusion p rotein GST-NC was 1 mg of infected leaves or 84 ng of purified virus. The metho d of dot hybridization using NC, a DIG-labelled DNA probe was 50 μg diseased l e aves, or 6 ng purified preparations. The detection endpoint of RT-PCR was 10 μg diseased leaves, or 2 ng purified virus preparation. Comparisons of sensitivit y and maneuverability were made among these methods.

The ecosystem characteristics of soil microorganism and the nutrient uptake of irrigated rice were investigated in a split-block experiment with different fertilization treatments, including control (no fertilizer application), PK, NK, NP, NPK fertilization, in the main block, and conventional rice and hybrid rice comparison, in the sub block. Average data of five treatments in five years indicated that the indigenous N supply (INS) capacity ranged from 32.72 to 93.21 kg/ha; that indigenous P supply (IPS)capacity ranged from 7.42 to 32.25 kg/ha; and that indigenous K supply (IKS) capacity ranged from 16.24 to 140.51 kg/ha, which showed that soil available nutrient pool depletion might occur very fast and that P, K deficiency has become a constraint to increasing yields of consecutive crops grown without fertilizer application. It was found that soil nutrient deficiency and unbalanced fertilization to rice crop had negative effect on the diversity of the microbial community and total microbial biomass in the soil.The long-term fertilizer experiment (LTFE) also showed that balanced application of N, P and K promoted microbial biomass growth and improvement of community composition. Unbalanced fertilization reduced microbial N and increased C/N ratio of the microbial biomass. Compared with inbred rice, hybrid rice behavior is characterized by physiological advantage in nutrient uptake and lower internal K use efficiency.

BACKGROUND:Since cannulated screw has been applied to femoral neck fracture, it is not uncommon that the screw wear penetrates or refunds. What factors affect the stability of cannulated screw for treatment of femoral neck fractures in adults remains unclear. ＜br> OBJECTIVE:To explore factors related to internal fixation failure by cannulated screws in treatment of adult femoral neck fracture and improve the stability of the adult femoral neck fracture by cannulated screws. ＜br> METHODS:A total of 92 adult patients of femoral neck fracture were treated by cannulated screws in our department between June 2007 and June 2011. Their data were retrospectively analyzed. According to clinical information and fol ow-ups, we selected factors such as age, gender, Garden type of fracture, preoperative skeletal traction, timing of surgery, Garden index, standards of pedicle screws, pedicle screw shapes, partial weight bearing time and postoperative complications, which may affect the success rate of cannulated screws for ＜br> treating femoral neck fracture. The selected factors were then grouped and assigned, after unrelated factors were excluded by one-way χ2 analysis, multiariable Logistic regression analysis was performed. ＜br> RESULTS AND CONCLUSION:The involved 92 patients were fol owed up for 18-72 months. According to Harris assessment criteria, hip function was excellent in 28 cases, good in 25 cases, fair in 17 cases, and poor in 22 cases at the final fol ow-up, the excellent and good rate was 58%. Radiographic results showed that, the patients were divided into two groups according to the presence of the displacement, GardenⅠ (n=22) and GardenⅡ (n=29) as a group, and Garden Ⅲ (n=25) and Garden Ⅳ (n=16) as the other group, the fixation failure rate was 12%and 39%, respectively. In normol and abnormal Garden Index groups, the fixation failure rate was 16%and 59%, respectively. In nail position standards and non-attainment standards groups, the fixation failure rate was 19%and 70%, respectively. In the complication and non-complication groups, the fixation failure rate was 14%and 55%, respectively. These factor groups showed significant differences (P＜0.05). Multiariable Logistic regression analysis showed that, Garden type of fracture, Garden index, standards of pedicle screws, and postoperative complications are the risk factors for internal fixation failure using cannulated screws in treatment of the adult femoral neck fracture.

BACKGROUND:There are few reports concerning effects of warming the yang and benefiting the marrow for the knee osteoarthritis on the expression of matrix metal oproteinase.
OBJECTIVE:To observe the effect of warming the yang and benefiting the marrow on the expression of matrix metal oproteinase in rabbit models of knee osteoarthritis.
METHODS:Of 96 healthy adult New Zealand rabbits, 72 rabbits were randomly selected for making rabbit models of knee osteoarthritis using plaster external fixation. After success model establishment, the rabbits were randomly assigned to three groups. Model group was left intact. Chinese medicine group received daily intragastric administration of drug extract 24 mL/kg. Drug control group was daily intragastrical y administered Puli Capsule (glucosamine hydrochloride) 24 mg/kg, once a day, until the eighth week of success model induction. An additional 24 New Zealand rabbits served as blank controls.
RESULTS AND CONCLUSION:Using quantitative PCR, matrix metal oproteinase-1, matrix metal oproteinase-3 and matrix metal oproteinase-13 expression was significantly higher in the model group than that in the other three groups. Matrix metal oproteinase-1, matrix metal oproteinase-3 and matrix metal oproteinase-13 expression was significantly lower in the Chinese medicine group and drug control group than that in the model group. These results indicated that warming the yang and benefiting the marrow for knee osteoarthritis in rabbits could effectively inhibit the expression of matrix metal oproteinase in rabbits.

Objective: To identify the functional domain of ?-Synuclein in affecting mitochondrial function and how the function to be impaired,especially,the mitochondrial membrane potential and the release of Cytochrome c.Methods: Harvest of ?-Syn-N and ?-Syn-△N by PCR,then subcloned into the pCMV-Myc mammalian expression vector.The recombinant plasmids were transfected into HEK293T cells by Lipofectamine 2000.After detecting the protein expression by Western blot,the functional domain was detected by co-immunoprecipitation.The mitochondrial membrane potential through flow cytometry and immunofluorescence,at the same time,the release of Cytochrome c through flow cytometry to detect.Results: The recombinant plasmids were constructed successfully.CO-IP has proved that N-terminal may be the functional domain of ?-Synuclein in affecting mitochondria.Over-expression of N-terminal could depolarize the mitochondrial membrane potential and induce the Cytochrome c releasing in MN9D cells.Conclusion: N-terminal may be the functional domain of ?-synuclein and over-expression of N-terminal could decrease mitochondrial activity.

Objective To evaluate the clinical significance of serum levels of ProGRP, TPS and NSE in diagnosis and therapy monitoring in small cell lung cancer patients. Methods The levels of serum ProGRP, TPS and NSE in 51 SCLC patients (SCLC group), 60 benign pulmonary disease patients (benign disease group ) and 60 healthy people (healthy group ) were determined using chemiluminescent immunoassay, ELISA and electrochemiluminescent immunoassay respectively. Blood samples were collected and detected prior to therapy, before the second course of chemotherapy and the third course of chemotherapy consecutively in all the 51 SCLC patients. Results The serum ProGRP, TPS and NSE concentrations prior to chemotherapy in limited stage SCLC (LSCLC) were 136. 9(22.8-631.7)ng/L, 78. 2(56.4-114.6) U/L and 28.1(20.9-46.1)μg/L, respectively; And in extensive stage SCLC patients (ESCLC) were 1 106.6(41.2-2161.1) ng/L, 230. 9( 143.5-259.0) U/L and 81.1 (34.3-140.0)μg/L, respectively. The serum concentrations of the 3 markers in benign disease group were 19. 7 ( 9. 5-29. 1 )ng/L, 48. 7 ( 17.9-95.4) U/L and 12. 1(1.2-13.9) μg/L; and in healthy group were 20.3(10.7-30.6) ng/L, 50.3(19.5-70.7) U/L and 11.7 (1.1-13.4)μg/L, respectively. The Kruskal-Wallis test showed significantly statistical difference in different groups of the 3 tumor markers, Chi-Square were 51. 368,36. 532 and 81. 645( P ＜0. 01 ). Significant statistically differences showed when the concentrations of the 3 marks of the 2 control group were compared with that of the LSCLC group ( U =491, 827, 609 and 476, 831, 585,respectively, P ＜ 0. 05 ). Differences were also statistically significant when the 2 control group compared with that of the ESCLC group ( U = 314,532,456 and 302,553,430, respectively, P ＜ 0. 01 ). The AUC of ProGRP was 0.832 +0.029(95% CI:0.774-0.890). When cutoff value of ProGRP set as 37.7 ng/L, the diagnostic sensitivity, specificity, positive predictive value, negative predictive value and Youden's index were 71% (36/51), 97% (116/120), 90% (36/40), 89% ( 116/131 ) and 67%, respectively; show good detection performance. The sensitivity increased to 92%, 86%, 92% and 88%, when combination detection of ProGRP + TPS + NSE, ProGRP + TPS, ProGRP + NSE and TPS + NSE were used, and the specificities were 77%, 77% , 92% and 77% accordingly. The Fridman test showed significantly statistical difference in the 3 tumor markers at different stages of treatment, x2 were 49. 120, 10. 614 and 44. 392, P ＜0. 01. After the first chemotherapy course, all the tumor marker levels except TPS decreased significantly in comparison with the pretreatment concentrations. However, only ProGRP levels showed a progressive drop during the two consecutive courses of therapy, and the median concentrations were 68.0 ( 18. 6-158.4 ) and 21.0( 14. 9-63.5) ng/L (compared to the level before therapy,Z=-4. 889 and -5. 594, P ＜0. 01 ). The median of serum TPS increased slightly to 105.2 (54. 1-181.2 ) U/L after the first chemotherapy course (Z=-1.248, P＞0.05), and decreased significantly to 79.0(48.7-155.3) U/L after the second chemotherapy course (Z=-2.484, P＜0. 05 ). As to the NSE, the median concentration decreased to 11.8(8.0-16.0)μg/L after the first chemotherapy course ( Z= - 5. 568, P ＜ 0. 01 ). However, the median was 10. 6(9.0-12.7)μg/L, which showed no significant decrease after the second chemotherapy course (Z=-1.851, P＞0.05).Forty-six SCLC patients evaluated as clinical remission ( 3 CR and 43 PR) after the second chemotherapy course, among them there were 38 patients (83%) with normal serum ProGRP, TPS and NSE level ( 19 patients) or with only 1 abnormal tumor level ( 19 patients). There were only 2 patients with all abnormal serum ProGRP, TPS and NSE level, and both patients were evaluated as clinical PD. Two patients with 2 abnormal tumors results were classified as SD, the only 1 patient without therapy evaluation also had 2 abnormal tumor marker results. Conclusions The serum ProGRP, TPS and NSE are valuable tumor markers for diagnosis and treat monitoring of SCLC, particularly the ProGRP + NSE shows the highest clinical value. Combing detection of the 3 tumor markers are valuable for therapy monitoring and prognosis in SCLC patients.

Limulus Factor C, a serine protease zymogen from the amoebocytes of the limulus, has high affinity for endotoxin. When Factor C is activated by endotoxin, it hydrolyses artificial tripeptide substrate and measurable products are released, so it can be used as an alternative reagent for endotoxin analysis. Factor C gene of Tachypleus tridentatus was obtained through RT-PCR and the recombinant protein was expressed by Bac-to-Bac/BmNPV baculovirus expression system in silkworm larvae. The activity of Factor C was detected with diluted serum of silkworm larvae, and the sensitivity of endotoxin detected was 0.2 EU/mL when the serum was diluted at 1:500. The silkworm larvae expressed limulus Factor C could be used to develop a new low-cost endotoxin test reagent.
Animals ; Arthropod Proteins ; biosynthesis ; Baculoviridae ; Bombyx ; metabolism ; Enzyme Precursors ; biosynthesis ; Genetic Vectors ; Larva ; metabolism ; Recombinant Proteins ; biosynthesis ; Serine Endopeptidases ; biosynthesis