To establish a cell model in vitro that stable expressing HBV by integrating HBA1.3 DNA into cell chromosome. The HBV1.3 full-length DNA was obtained by digested pGEM-HBV1.3 plasmid with HindIII and then was linked with PU-21 vector digested by HindIII. This was resulted in generation of a recombined plasmid named PU21-HBV plasmid. The recombined plasmid was introduced into HepG2 cells by electroporation. The transfected cells were screened with G418. The insertion and expression of HBV were identified by X-gal staining, RT-PCR and Southern blot. The result of PU21-HBV plasmid sequence demonstrated that HBV1.3 DNA was linked correctly with PU-21 vector. A series of positive cell colonies were obtained with G418 screening followed transfecting PU21-HBV plasmid into HepG2 cells. The results of Southern blot and RT-PCR exhibit that HBV1.3 DNA had successfully integrated into the chromosomes of HepG2 cells and had functional HBV gene transcription. HBV1.3 DNA was inserted into HepG2 genome and could stable transcript HBV RNA. The stable HBV expression cell line was constructed successfully. There are LoxP sites in the trapping vector PU21. With the Cre enzyme, interesting genes could be excganged into the LoxP sites. Therefore, double stable expression of interesting gene and HBV cell lines could be generated. The cell lines will be useful for further research some target gene function on replication of HBV.

Under indoors simulating natural growing condition, the 4th-instar Mythimna separata larvae were fed by using poi- son leaf disk method. The effect of total ginsenosides on the protective enzymes (PPO, T-SOD, CAT and POD) of M. separata larvae was studied. The total ginsenosides could influence the protective enzymes of 4th-instar M. separata larvae significantly. After treated by total ginsenosides, the PPO activities increased firstly then decreased, and tended to equilibrium, and reached the maximum after 48 h. Furthermore, the total ginsenosides disturbed the dynamic balance of SOD, CAT and POD of M. separata larvae, and the yield of O2-* speeded. The results suggest that the total ginsenosides influence the protective enzymes of 4th-instar M. separata larvae, and disturb the original dynamic balance of protective enzymes. Consequently the insect suffers from the harm of O2-*.
Animals ; Enzymes ; metabolism ; Ginsenosides ; metabolism ; Larva ; metabolism ; Lepidoptera ; metabolism ; Oxygen ; metabolism

This paper is in order to study the anti-feeding and growth inhibition activity of toatal ginsenoside of ginseng stems and leaves against 4th-instar Mythimna separata larvae. Simulating natural growing condition indoors, on the base, To study the anti-feeding and growth inhibition activity of toatal ginsenoside against 4th-instar M. separata larvae by leaf disc test. The toatal ginsenoside appeared to be of significant antifeeding activity against 4th-instar M. separata larvae. The 4th-instar M. separata larvae fed on the leaves of Sorghum bicolor treated with 20, 10, 5 g · L(-1) toatal ginsenoside. At 8 h, non-selective anti-feeding rate were 88.67%, 64.40% and 47.36%, and selective anti-feeding rate were 62.49% , 44.29% and 34.19%; Compared with the photographic, The toatal ginsenoside conld make the development period had prolonged 13h in treated group. The toatal ginsenoside had significant inhibition effect on feeding and growth and development against 4th-instar M. separata larvae, and inhibition effect increases as the increase of concentration ginsenoside.
Animals ; Ginsenosides ; pharmacology ; Insecticides ; pharmacology ; Larva ; Moths ; growth & development ; Panax ; chemistry

OBJECTIVETo observe the therapeutic effect of acupuncture on gastrointestinal dysmotility in liver cirrhosis.

METHODSForty cases of gastrointestinal dysmotility in liver cirrhosis were randomized into an acupuncture group and a motilium group, 20 cases in each one. In the acupuncture group, on the basis of the conventional treatment, electroacupuncture was applied at Zusanli (ST 36), Sanyinjiao (SP 6) and Taichong (LR 3) for 30 min, once a day. In the motilium group, on the basis of the conventional treatment, motilium was taken orally 30 min before meals, 10 mg each time, three times a day. The treatment was required for 2 weeks in both groups. The changes in the digestive tract symptom score and liver function Child-Pugh score were observed and the efficacy was assessed.

RESULTSThe total effective rate of digestive tract symptoms was 85.0% (17/20) in the acupuncture group and 70.0% (14/20) in the motilium group. The score improvements in abdominal distention, belching and vomiting in the acupuncture group were superior to those in the motilium group (all P < 0.05). In the acupuncture group, the liver function Child-Pugh total score was 8.40 +/- 0.22 before treatment and reduced to 5.36 +/- 0.17 after treatment, in which the scores for ascites, serum bilirubin and albumin were all reduced (all P < 0.05) and the reducing range was increased in tendency with the improvements in digestive tract symptoms. In the motilium group, Child-Pugh score was not changed obviously as compared with that before treatment. CONCLUSION Acupuncture effectively alleviates digestive tract symptoms and improves liver function for the patients of liver cirrhosis, its efficacy on gastrointestinal dysmotility in liver cirrhosis is superior to motilium.

Acupuncture Therapy ; Adult ; Aged ; Domperidone ; therapeutic use ; Female ; Gastrointestinal Motility ; drug effects ; Humans ; Liver Cirrhosis ; drug therapy ; physiopathology ; therapy ; Male ; Middle Aged ; Treatment Outcome ; Young Adult

OBJECTIVETo screen the potential protein biomarkers in minimal residual disease (MRD) of the acute promyelocytic leukemia (APL) by comparison of differentially expressed serum protein between APL patients at diagnosis and after complete remission (CR) and healthy controls, and to establish and verify a diagnostic model.

METHODSSerum proteins from 36 cases of primary APL, 29 cases of APL during complete remission and 32 healthy controls were purified by magnetic beads and then analyzed by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS). The spectra were analyzed statistically using FlexAnalysis(TM) and ClinProt(TM) software.

RESULTSTwo prediction model of primary APL/healthy control, primary APL/APL CR were developed. Thirty four statistically significant peptide peaks were obtained with the m/z value ranging from 1000 to 10 000 (P < 0.001) in primary APL/healthy control model. Seven statistically significant peptide peaks were obtained in primary APL/APL CR model (P < 0.001). Comparison of the protein profiles between the two models, three peptides with m/z 4642, 7764 and 9289 were considered as the protein biomarker of APL MRD. A diagnostic pattern for APL CR using m/z 4642 and 9289 was established. Blind validation yielded correct classification of 6 out of 8 cases.

CONCLUSIONSThe MALDI-TOF MS analysis of APL patients serum protein can be used as a promising dynamic method for MRD detection and the two peptides with m/z 4642 and 9289 may be better biomarkers.

Adolescent ; Adult ; Aged ; Case-Control Studies ; Child ; Humans ; Leukemia, Promyelocytic, Acute ; classification ; diagnosis ; Male ; Middle Aged ; Neoplasm, Residual ; classification ; diagnosis ; Prognosis ; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ; methods ; Young Adult

Bronchoalveolar lavage is an important animal experimental technique in the study of respiratory system and its pathological changes. It can acquire a variety of biochemical factors, inflammatory mediators and immune cells from the respiratory tract and lungs, and provides an important evaluation index and reference for animal experiment. Bronchoalveolar lavage is an effective and reliable method for the diagnosis of respiratory diseases. It has been gradually standardized and widely used in clinical practice at present, however, there is no set of standard for bronchoalveolar lavage in rats and mice. The results of bronchoalveolar lavage fluid are affected by many factors, such as the lavage fluid, suction pressure, the amount of lavage and recovery, and the retention time of lavage fluid in the lungs. Successful and efficient acquisition of lavage specimens is the key to the study and evaluation of respiratory diseases. This paper summarizes the current lavage methods commonly used by domestic and foreign researchers, and provides a reference for further research in the this field.

OBJECTIVE To explore the mechanisms of the volatiles of Wendan granule for the treatment of senile dementia,network pharmacology method integrating absorption,distribution,metab-olism, and excretion (ADME) screening, target fishing, network constructing, pathway analyzing, and correlated diseases prediction was applied. METHODS Twelve small molecular compounds of WDG were selected as the objects from 74 volatiles with the relative abundances above 2%,and their ADME parameters were collected from Traditional Chinese Medicine Systems Pharmacology platform (TCMSP), and then the corresponding targets, genes, pathways and diseases were predicted according to the data provided by TCMSP,DrugBank,Uniport and the Database for Annotation,Visualization and Integrated Discovery(DAVID).The related pathways and correlation analysis were explored by the Kyoto Encyclo-pedia and Genomes (KEGG) database. Finally, the networks of compound-target, target-pathway and pathway-disease of WDG were constructed by Cytoscape software. RESULTS Twelve compounds interacted with 49 targets, of which top three targets were Gamma-aminobutyric acid receptor subunit alpha-1 (GABRA1), Prostaglandin G/H synthase 2 (PGHS-2) and Sodium-dependent noradrenaline transporter.Interestingly,these targets were highly associated with depression,insomnia and Alzheimer′s disease that mainly corresponded to mental and emotional illnesses. CONCLUSION The integrated network pharmacology method provides precise probe to illuminate the molecular mechanisms of volatiles of WDG for relieving senile dementia related syndromes,which will also facilitate the application of traditional Chinese medicine in modern medicine,as well as follow-up studies such as upgrading the quality stan-dard of clinical medicine and novel drug development.

OBJECTIVETo develop quality of life questionnaire of Chinese medicine for postoperative patients with colorectal cancer (QLQ-CMPPCC), thus comprehensively and objectively evaluating the clinical efficacy of Chinese medicine and pharmacy in treating postoperative patients with colorectal cancer (CC).

METHODSThe theoretical structure model of the questionnaire was addressed in combined with basic theories of Chinese medicine according to the principle of WHO quality of life (QOL). The primary questionnaire was developed using methods of structuralization policy making after we extensively retrieve various universal and specific questionnaires for CC cancer patients at home and abroad. The 205 CC patients were tested by questionnaire. The items were screened using experts grading method, item selection analysis, dispersion trends of standard deviation, t-test, correlation coefficient method, factor analysis,and Cronbach's alpha.

RESULTSThe QLQ-CMPPCC was developed containing four domains of physical, psychological, independence, and social functions, involving 20 aspects and 54 items. Of them, non-fistula patients answered 43 items and fistula patients answered 46 items. One item covered the general QOL evaluation.

CONCLUSIONSQLQ-CMPPCC showed Chinese medical features. It comprehensively reflected the connotation of QOL for postoperative CC patients. It could be taken as a tool for evaluating Chinese medical efficacy for postoperative CC patients.

Colorectal Neoplasms ; surgery ; Humans ; Medicine, Chinese Traditional ; methods ; Postoperative Period ; Quality of Life ; Surveys and Questionnaires ; Treatment Outcome

OBJECTIVETo evaluate the disadvantage of nonstandard operation for thyroid papillary cancer, and the value of re-operation.

METHODSEight hundred and ninety three thyroid papillary cancer patients (332 patients who received nonstandard operation received re-operation and 561 patients who received first standard operation in Zhejiang Cancer Hospital from January 1990 to January 2000) were retrospectively reviewed.

RESULTSPathological results confirmed that there were 53.9% cases with residual cancer in re-operative specimen. The parathyroid was preserved in 74.1% cases in re-operation and 93.0% cases in standard operation (P < 0.01). The recurrent laryngeal nerve was injured in 3.3% cases in reoperation and 1.2% cases in standard operation (P < 0.05). The positive neck lymph node was 39.2% and 37.4% (P > 0.05), the 5-year local recurrence rate was 7.5% and 3.7% (P < 0.01), the total 5-year and 10-year cum-survival rates were 90.2%, 84.4% in patients with re-operation and 94.0%, 92.5% in patients with standard operation. The 10-year cum-survival rate were 67. 8% in patients with recurrence and 92. 9% in patients with non-recurrence in primary site respectively. Statistical analysis showed that the cancer recurrence influences the survival rate significantly (P < 0.01).

CONCLUSIONSIn consideration of the higher residual tumor rate in patients who received nonstandard operations, the re-operations were necessary. But the re-operation could also leads to higher recurrence rate and more functional injuries. The standardized primary operation should therefore be formulated and advocated.

Adolescent ; Adult ; Aged ; Carcinoma, Papillary ; surgery ; Child ; Female ; Humans ; Male ; Middle Aged ; Reoperation ; Thyroid Neoplasms ; surgery ; Thyroidectomy ; methods ; Young Adult

OBJECTIVEThis study aims to reveal the effect of total ginsenoside on the protein content and digestive enzyme activities of 4th-instar Mythimna separata larvae, including alpha-amylase and cellulose, and explore the ecological function of total ginsenoside.

METHODWhile simulating natural growing condition indoors, 4th-instar M. separata larvae were fed by poison leaf disk method. The protein content was tested by Lowry Protein Assay Kit method, the activity of alpha-amylase was measured by dinitrosalicylic acid test, and the activity of cellulase was determined by the filter paper method.

RESULTThe total ginsenoside could reduce the content of protein of 4th-instar M. separata larvae significantly, and the activity of digestive enzyme, including alpha-amylase and cellulase. The protein content, alpha-amylase and cellulase activity of treatments were obviously lower than that of the control. Inhibition ratio of alpha-amylase and cellulase activity was positively correlated with total ginsenoside concentration: i. e. 20 g x L(-1) > 10 g x L(-1) > 5 g x L(-1).

CONCLUSIONThe results suggest that the inhibition effect of total ginsenoside on protein content and digestive enzymes may be one of the causes to antifeedant and dysplasia of M. separata larvae.

Animals ; Digestion ; Ginsenosides ; pharmacology ; Insect Proteins ; metabolism ; Larva ; drug effects ; enzymology ; growth & development ; Moths ; drug effects ; enzymology ; growth & development