Objective To optimize the conditions of process for extraction of volatile oil of Rhizoma Zedoariae (RZ) in Weishu Granules and preparation of its ? - cyclodextrin- inclusion complexes. Methods Orthogonal design was adopted. Extraction content of RZ volatile oil was used as the index for the optimizing of extraction process. The amount of inclusion complexes and efficient inclusion rate served as the indexes for the optimizing of inclusion process. Ultraviolet spectrophotometry was applied to detect the content of volatile oil. Results The optimal extraction conditions were: gringding the crude RZ into coarse grains, adding 12- fold water and distilling the mixture for 6 h; the optimal inclusion conditions were: ? - cyclodextrin and volatile oil were mixed in the proportion of 8 :1, 2- fold water was added into the mixture and the mixture was triturated for 60 min. The maximum UV spectrum absorbance of aethylis acetas liquor of valatile oil located at wavelength of 253nm. Conclusion Higher extraction rate of RZ volatile oil can be achieved by this technique; volatile oil is powdered and its stability increased after being included by ? - cyclodextrin.

Total mRNA was extracted from lymphocytes separated from the peripheral blood of allergic patients, and then variable region of heavy chain (VH) and variable region of light chain (VL) cDNA library were constructed by RT-PCR. Human scFv templates for rabbit reticulocyte lysate ribosome display were assembled by primers and linker peptide (Gly4Ser)3. mRNA bound in antibody-ribosome-mRNA complexes was recovered using in-situ single primer RT-PCR, and three rounds of anti-IgE scFv DNA were enriched. The target DNA fragments were double enzyme digested and ligated into plasmid pET22b (+), followed by transformation in E. coli Rosseta (DE3). Positive clones were screened using clone PCR, Dot blotting and antigen ELISA. The correct lengths of VH (400 bp) and VL (710 bp) PCR products were obtained. The expected 1,000 bp ribosome display templates were also observed in agarose gel electrophoresis. After three rounds of ribosome display target sequences were effectively enriched, leading to a library of 10(13) members. Antibodies with the highest ELISA value for IgE were generated in the strain pET-IgE-6. A human anti-IgE scFv library was successfully constructed as described herein. Ribosome display using single primer in-situ RT-PCR as the recovery procedure effectively enriched target sequences. Anti-IgE scFv with high affinity and specificity were identified. The prepared human anti-IgE scFv fragment might be self-developed to a lead drug for treating asthma. Our study provides an alternative method for rapid discovery of human antibodies of therapeutic importance.

Total mRNA was extracted from lymphocytes separated from the peripheral blood of allergic patients, and then variable region of heavy chain (VH) and variable region of light chain (VL) cDNA library were constructed by RT-PCR. Human scFv templates for rabbit reticulocyte lysate ribosome display were assembled by primers and linker peptide (Gly4Ser)3. mRNA bound in antibody-ribosome-mRNA complexes was recovered using in-situ single primer RT-PCR, and three rounds of anti-IgE scFv DNA were enriched. The target DNA fragments were double enzyme digested and ligated into plasmid pET22b (+), followed by transformation in E. coli Rosseta (DE3). Positive clones were screened using clone PCR, Dot blotting and antigen ELISA. The correct lengths of VH (400 bp) and VL (710 bp) PCR products were obtained. The expected 1,000 bp ribosome display templates were also observed in agarose gel electrophoresis. After three rounds of ribosome display target sequences were effectively enriched, leading to a library of 10(13) members. Antibodies with the highest ELISA value for IgE were generated in the strain pET-IgE-6. A human anti-IgE scFv library was successfully constructed as described herein. Ribosome display using single primer in-situ RT-PCR as the recovery procedure effectively enriched target sequences. Anti-IgE scFv with high affinity and specificity were identified. The prepared human anti-IgE scFv fragment might be self-developed to a lead drug for treating asthma. Our study provides an alternative method for rapid discovery of human antibodies of therapeutic importance.
Amino Acid Sequence ; Antibodies, Anti-Idiotypic ; genetics ; isolation & purification ; Antibody Affinity ; Asthma ; blood ; Base Sequence ; DNA, Complementary ; metabolism ; Escherichia coli ; metabolism ; Humans ; Immunoglobulin Heavy Chains ; genetics ; Immunoglobulin Light Chains ; genetics ; Immunoglobulin Variable Region ; genetics ; Lymphocytes ; chemistry ; Peptide Library ; RNA, Messenger ; isolation & purification ; Recombination, Genetic ; genetics ; Ribosomes ; chemistry ; genetics ; immunology ; Single-Chain Antibodies ; genetics ; isolation & purification ; Transformation, Genetic

This study aims at trying to establish a novel method of sterility test for injections based on biothermodynamics, in order to overcome the deficiencies of routine sterility tests such as long detecting cycle, low sensitivity and prone to misjudgments. A biothermodynamics method was adopted to rapidly detect the microorganism contamination of injections by monitoring the heat metabolism during the growth of microbe. The growth rate equal to or greater than zero and the heat power difference of P(i) and P(0) with three folds higher than the noise of baseline were chosen as indexes to study the heat change rule of microbe. In this way, the effectiveness of the new method to detect strains required by conventional sterility test or in injection samples was also investigated. Results showed that the Gram-positive bacteria, Gram-negative bacteria and fungi demanded by sterility testing methodology could be detected by biothermodynamics method within 10 hours, with the sensitivity lower than 100 CFU x mL(-1). Meanwhile, this method was successfully applied to the sterility test of Compound Yinchen injection (FFYC), Shuanghuanglian powder injection (SHL) and Compound Triamcinolone injection (TAND) which were sterilized with different degrees. Therefore, the biothermodynamics method, with advantages of fast detection and high sensitivity, could be a complementary solution for conventional sterility tests.
Anti-Inflammatory Agents ; administration & dosage ; chemistry ; Drug Contamination ; Drugs, Chinese Herbal ; administration & dosage ; chemistry ; Fungi ; isolation & purification ; Gram-Negative Bacteria ; isolation & purification ; Gram-Positive Bacteria ; isolation & purification ; Hot Temperature ; Injections ; Microbiological Techniques ; methods ; Sensitivity and Specificity ; Sterilization ; Triamcinolone ; administration & dosage ; chemistry

OBJECTIVETo study the clinical characteristics of acute myocardial infarction (AMI) among younger adults and to explore the possible mechanisms of early myocardial infarction, combined with the newly discovered risk factors of coronary heart disease.

METHODSData on comparative analysis to the exposure rates of the risk factors and inducing factors of non-CAD patients with two groups of AMI patients including younger adults group (< or =40 years old) and aged adults group (> or =50 years old). Coronary angiography was applied.

RESULTSThere were differences noticed between the frequencies of risk factors of the two AMI groups. In younger adults group the exposure rates of smoking, hyperlipidemia, positive family history, C-reactive protein (CRP) and fibrinogen were markedly higher, while in elderly group the exposure rates of hypertension, smoking, hyperlipidemia, diabetes, CRP, fibrinogen and homocysteine (HCY) were markedly higher (P < 0.05). Although the clustering status of risk factors of the younger adult group was not higher than that of the elderly group. There were obvious inducing factors before the patients were attacked by AMI and the inducing factors inclined to cluster, which had obvious dose-reaction relationships with the occurrence of AMI in young people.

CONCLUSIONEarly AMI of younger adults might relate to the clustering status of inducing factors. The coexistence of several kinds of inducing factors was resulted in the occurrence of AMI of the atherosclerosis (As) and non-As patients by means of myocardial ischemia accumulation effect.

Adult ; Age Factors ; Aged ; Atherosclerosis ; epidemiology ; China ; epidemiology ; Coronary Angiography ; Humans ; Middle Aged ; Myocardial Infarction ; epidemiology ; pathology ; Myocardial Ischemia ; epidemiology ; Risk Factors

Objective To analyze the correlation between clinical mental personality,coping style,social support and depression,anxiety for patients with chronic hepatitis B,and their treatment strategies.Methods A total of 66 patients with chronic hepatitis B (CHB group) and 72 normal people (control group) were investigated by the questionnaire,such as the demographic history questionnaire,Eysenck Personality Questionnaire (EPQ),Trait Coping Style Questionnaire (TCSQ),Perceived Social Support Scale (PSSS),Hamilton Depression Scale (HAMD),Hamilton Anxiety Scale (HAMD) were assessed.Results The HAMD score and HAMA score of hepatitis B group were significantly higher than that of the control group.Pearson correlation analysis showed that depressive symptoms was negatively correlated with positive coping,family support,extroversion,and was positively correlated with neuroticism.Multiple linear regression analysis showed family support,active coping,neuroticism and psychosis were all included in the regression equation and can predict depressive symptoms (3 values,0.002,0.003,0.004,0.022,respectively).Neuroticism,positive responded to the two independent variables (β values,0.003,0.011,respectively) into the regression equation,and can predict anxiety symptoms.Conclusion The patients with chronic hepatitis B were more prone to suffer from depression and anxiety than normal people,depressive symptoms are closely related to the neuroticism and neuroticism.The anxiety symptoms are related to neuroticism.The neurotic personality traits,lack of positive coping strategies and out-of-home support may lead to depression and anxiety.Depression and anxiety symptoms can be avoided by improving the patient's personality characteristics,and actively responding and enhancing the social support system outside the home soas to reduce depression,anxiety symptoms in patients with hepatitis B.

Objective To analyze the correlation between clinical mental personality,coping style,social support and depression,anxiety for patients with chronic hepatitis B,and their treatment strategies.Methods A total of 66 patients with chronic hepatitis B (CHB group) and 72 normal people (control group) were investigated by the questionnaire,such as the demographic history questionnaire,Eysenck Personality Questionnaire (EPQ),Trait Coping Style Questionnaire (TCSQ),Perceived Social Support Scale (PSSS),Hamilton Depression Scale (HAMD),Hamilton Anxiety Scale (HAMD) were assessed.Results The HAMD score and HAMA score of hepatitis B group were significantly higher than that of the control group.Pearson correlation analysis showed that depressive symptoms was negatively correlated with positive coping,family support,extroversion,and was positively correlated with neuroticism.Multiple linear regression analysis showed family support,active coping,neuroticism and psychosis were all included in the regression equation and can predict depressive symptoms (3 values,0.002,0.003,0.004,0.022,respectively).Neuroticism,positive responded to the two independent variables (β values,0.003,0.011,respectively) into the regression equation,and can predict anxiety symptoms.Conclusion The patients with chronic hepatitis B were more prone to suffer from depression and anxiety than normal people,depressive symptoms are closely related to the neuroticism and neuroticism.The anxiety symptoms are related to neuroticism.The neurotic personality traits,lack of positive coping strategies and out-of-home support may lead to depression and anxiety.Depression and anxiety symptoms can be avoided by improving the patient's personality characteristics,and actively responding and enhancing the social support system outside the home soas to reduce depression,anxiety symptoms in patients with hepatitis B.

OBJECTIVETo investigate the correlation of radiation pneumonitis (RP) with standardized uptake value (SUV) for fluorodeoxyglucose (FDG) positron emission tomography and computed tomography (PET-CT) in lung cancer patients treated with radiation therapy.

METHODSFourty patients with unresectable non-small cell lung cancer (NSCLC) received FDG PET-CT before and after radiotherapy. The average SUV of the lung tissue irradiated with a dose of < or = 5 Gy, 5.1 approximately 15 Gy, 15.1 approximately 35 Gy, 35.1 approximately 60 Gy, >60 Gy were measured. The correlation between SUV and RP was analyzed by comparing the SUV in the patients with RP and without. The SUV ratio of the irradiated lung tissue to that of the non-irradiated lung tissue (L/B) was also calculated.

RESULTSOf the 40 patients, 8 developed RP, including 6 cases of grade 2 and 2 cases of grade 3. The SUV of irradiated lung tissues with a dose of 35.1 approximately 60 Gy was significantly correlated with RP. When SUV > or =1, the RP incidence rate was 41.7% versus 20.0% in the whole group, with a statistically significant difference. (chi2 = 3.96, P < 0.05). The sensitivity and specificity of SUV in predicting RP was 62.5% and 78.1%, respectively. When the value of L/B > or = 2.5, the RP incidence rate was 40.7% in this group versus 20.0% in the whole group, with a statistical significance (chi(2) = 4.92, P < 0.05). If taking L/B > or = 2.5 as a threshold value, the sensitivity and specificity in predicting RP was 72.7% and 90.9%, respectively. No statistically significant difference was found in predicting radiation pneumonitis between SUV > or =1 and L/B > or = 2.5 (chi2 = 0.002, P > 0.05).

CONCLUSIONThe standardized uptake value (SUV) and the SUV ratio of the irradiated lung tissue to that of the non-irradiated lung tissue (L/B) for FDG PET-CT are positively correlated with radiation pneumonitis, and clinicians may use it to predict the occurrence of radiation pneumonitis.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Non-Small-Cell Lung ; radiotherapy ; Female ; Fluorodeoxyglucose F18 ; pharmacokinetics ; Follow-Up Studies ; Humans ; Lung Neoplasms ; radiotherapy ; Male ; Middle Aged ; Positron-Emission Tomography ; Radiation Dosage ; Radiation Pneumonitis ; diagnosis ; etiology ; metabolism ; Radiopharmaceuticals ; pharmacokinetics ; Radiotherapy, Conformal ; adverse effects ; Tomography, X-Ray Computed

BACKGROUNDWhen performing superior vena cava isolation, the major concerns are inadvertent ablation on sinus node and right phrenic nerve. However, little is known about the spatial relationship of electrical connections between superior vena cava and right atrium with the sinus node and phrenic nerve locations among individual patients.

METHODSWe studied 87 patients (male/female 60/27, mean age of (51 +/- 9) years) with atrial fibrillation. Before superior vena cava isolation, the sinus node site was defined by right atrium activation mapping during sinus rhythm and the right phrenic nerve site was localized via pacing manoeuvre. Superior vena cava was isolated by ablation at the electrical connection under the guidance of circular mapping catheter. The sites of sinus node, phrenic nerve and electrical connections were noted. Continuous variables were compared using Student's t test. A P value < 0.05 was considered statistically significant.

RESULTSRight atrium activation mapping revealed that the sinus node located at the anterior lateral segment of superior vena cava-right atrium junction in all patients. In 82 patients with detectable diaphragmatic stimulations, the phrenic nerve sites were predominantly at the lateral segment (70/82) with anterior lateral and anterior segments for a few patients. A total of 165 electrical connections were located among all 87 patients, and this averaged 1.8 +/- 0.6 (1-3) per patient. The anterior septum (72 patients (43.6%)), the anterior wall (40 (24.2%)), and the posterior septum (35 (35.4%)) of superior vena cava-right atrium junction were the electrical connection regular sites. Superior vena cava was isolated in all patients. Two patients developed sinus bradycardia, with 3 mild superior vena cava stenosis and 2 phrenic nerve palsy.

CONCLUSIONSThe sinus node, phrenic nerve and electrical connection sites were distributed along the superior vena cava-right atrium junctions at expected locations for most patients. The electrical connections were separated from the sinus node and phrenic nerve sites. With the activation mapping of right atrium and pacing along superior vena cava-right atrium junctions, the sinus node and phrenic nerve were localized and superior vena cava isolated in most patients.

Adolescent ; Adult ; Aged ; Aged, 80 and over ; Atrial Fibrillation ; pathology ; surgery ; Catheter Ablation ; methods ; Echocardiography ; Electrophysiology ; Female ; Heart Atria ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Phrenic Nerve ; pathology ; surgery ; Prospective Studies ; Sinoatrial Node ; pathology ; surgery ; Vena Cava, Superior ; pathology ; Young Adult

OBJECTIVETo investigate the expression of KIAA0101 protein in gastric carcinoma cells, and to explore the effects of its down-regulation on the cell proliferation, cell cycle and invasion.

METHODSWestern blot was used to detect KIAA0101 protein expression in three gastric carcinoma cell lines including MKN-28, SGC-7901 and MKN-45. KIAA0101 siRNA and control siRNA were utilized to transfect MKN-45 cells, respectively. CCK-8 was used to analyze the changes of cell proliferation, and flow cytometry to examine the changes of cell cycle distribution. Finally, Boyden chamber was used to detect the ability of cell invasion.

RESULTSRelative level of KIAA0101 protein in MKN-45 cells was significantly higher than those in MKN-28 and SGC-7901 cells, and there was significant difference among the three cell lines (P < 0.05). The result of CCK-8 study demonstrated that, compared with untreated group and control siRNA group, the proliferation of MKN-45 cells in KIAA0101 siRNA group was significantly inhibited (P < 0.05). Additionally, the result of cell cycle analysis revealed that the percentage of cell number in G(0)/G(1) phase in KIAA0101 siRNA group [(61.47 ± 0.89)%] was significantly higher than those in untreated group [(47.43 ± 0.85)%] and control siRNA group [(48.43 ± 0.73)%; F = 271.653, P = 0.000]. Further, Boyden chamber assay showed that the cell numbers migrated to Matrigel in KIAA0101 siRNA group (61.51 ± 4.76) were significantly lower than those in untreated group (138.74 ± 10.16) and control siRNA group (132.93 ± 11.25; F = 65.949, P = 0.000).

CONCLUSIONSDown-regulation of KIAA0101 expression leads to an inhibition of cell proliferation, cell cycle and cell invasion. It may provide a novel target for the treatment of patients with gastric carcinoma.

Carrier Proteins ; genetics ; metabolism ; Cell Cycle ; Cell Line, Tumor ; Cell Movement ; Cell Proliferation ; Down-Regulation ; Gene Expression Regulation, Neoplastic ; Humans ; Neoplasm Invasiveness ; RNA Interference ; RNA, Small Interfering ; genetics ; Stomach Neoplasms ; metabolism ; pathology ; Transfection