With the application of monoclonal antibody technology more and more widely, its production technology is becoming more and more perfect. Small molecule monoclonal antibody technology is becoming a hot research topic for people. The application of traditional Chinese medicine small molecule monoclonal antibody technology has been more and more widely, the technology for effective Chinese medicine component knockout provide strong technical support. The preparation of monoclonal antibodies and small molecule knockout technology are reviewed in this paper. The preparation of several steps, such as: in the process of preparation of antigen, hapten carrier coupling, coupling ratio determination and identification of artificial antigen and establishment of animal immunization and hybridoma cell lines of monoclonal antibody, the large-scale preparation; small molecule monoclonal antibody on Immune in affinity chromatography column method is discussed in detail. The author believes that this technology will make the traditional Chinese medicine research on a higher level, and improve the level of internationalization of Chinese medicine research.
Animals ; Antibodies, Monoclonal ; chemistry ; genetics ; immunology ; Humans ; Hybridomas ; metabolism ; Immunologic Techniques ; methods ; trends

Objective:To analyze and compare the volatile components in vinegar-processed schisandrae sphenantherae fructus and vinegar-processed schisandrae chinesis fructus.Methods:The volatile components in vinegar-processed schisandrae sphenantherae fruc-tus and vinegar-processed schisandrae chinesis fructus were extracted by headspace solid phase-microextraction (HS-SPME) and quali-tatively analyzed by GC-MS.Results:Totally 20 kinds of constituents were identified from vinegar-processed schisandra sphenanthera fructus, which accounted for 99.55%of the total volatile components , and totally 21 kinds of constituents were identified from vinegar-processed schisandra sphenanthera fructus, which accounted for 99.90% of the total volatile components .Conclusion: The type and content of volatile components in vinegar-processed schisandrae sphenantherae fructus and vinegar-processed schisandrae chinesis fructus are quite different , and the study can provide scientific basis for the two traditional Chinese medicinal materials .

Objective To prepare hybrid substrate and apply it to detect thiram with surface-enhanced Raman spectros-copy( SERS) which provides unique molecular vibration information .Methods The Au substrate was prepared by deposi-tion of gold film on the silver substrate that had a rough surface .The Au substrate was treated with amination as a linker with the silver sol before the hybrid substrate was formed .With PATP as a probe molecule ,the Raman intensity of PATP on the Au substrate and the hybrid substrate was compared ,respectively .Results and Conclusion PATP had stronger Raman intensity on hybrid substrate than on the Au , and the detection limit was 10 -9 mol/L.This method can be used for quanti-tative detection on the hybrid substrate by SERS .

Objective To study the application of super-selective external carotid artery embolization in head and neck diseases. Methods DSA and super-selective external carotid artery embolization were carried out in 41 cases of head and neck diseases including 12 cases of epistaxis,7 nasopharyngeal fibroangioma,1 traumatic arterial bleeding,14 vascular malformation,and 7 malignancies. Results No recurrence of nose bleeding after embolization of epistaxis was seen within 6 - 12-month follow up. The operative bleeding was reduced significantly by preoperative embolization in nasopharyngeal fibroangioma. No recurrence of bleeding was achieved after embolization of traumatic artery. Among the cases of vascular malformation,3 were proven to be significantly effcient,6 effcient,and 5 inefficent in the 6 - 12-month follow up. Among the 7 malignant cases,3 survived more than 2 years. Conclusion Super-selective external carotid artery embolization is safe and effective in the treatment of head and neck diseases. (J Intervent Radiol,2006,15: 330-332)

Background:Diagnosis of liver cirrhosis in early stage with early intervention may stabilize disease progression, avoiding or delaying the occurrence of decompensation. Seeking non-invasive serum biomarkers is becoming an important topic in the diagnosis and assessment of liver cirrhosis. Aims:To study the value of serum miR-192 and miR-29a as non-invasive biomarkers for the diagnosis of liver cirrhosis. Methods:Differentially expressed serum miRNAs between patients with liver cirrhosis and healthy controls were screened through online literature retrieval and then confirmed by real-time PCR. Serum levels of two confirmed miRNAs,miR-192 and miR-29a were analyzed in 120 patients with liver cirrhosis and 76 healthy volunteers by real-time PCR. A mathematical model of combined detection of miR-192 and miR-29a for diagnosis of liver cirrhosis was established by binary logistic regression. The diagnostic performance of various non-invasive serum indicators was evaluated by ROC curve analysis. Results:Compared with healthy controls,expression level of serum miR-192 in cirrhotic patients was significantly increased and that of serum miR-29a was significantly reduced(P ﹤ 0. 001). The diagnostic performance of risk score obtained from mathematical model of combined detection of serum miR-192 and miR-29a was superior to that of single miRNA detection or other non-invasive serum indicators,such as APRI,FIB-4 and ARR,the areas under ROC curve of the above mentioned indicators were 0. 968,0. 887,0. 933,0. 796,0. 793,and 0. 571,respectively. Serum levels of miR-192,miR-29a and the risk score of their combined detection were significantly correlated with the stage of liver cirrhosis according to the Child-Pugh classification( P ﹤ 0. 05). Conclusions:Serum miR-192,miR-29a and the risk score of their combined detection might be novel non-invasive biomarkers for the diagnosis and assessment of liver cirrhosis.

Data mining technology has become a powerful tool in traditional Chinese medicine (TCM) research. In this paper, based on the principle and basic requirements of data mining, the mining methods and procedures were described. And then the application of data mining technology in Chinese medicine pair research was classified and summarized, such as the compatibility characters, characteristic pairs, dosage-effect relationship and property compatibility, which provide the direction and data base for modern research of Chinese medicine pair.
Cluster Analysis ; Data Mining ; methods ; Drug Interactions ; Drug Prescriptions ; Medicine, Chinese Traditional ; methods

Objective To compare the sub-cellular proteome of isoniazid ( INH)-resistant Mycobacterium tuberculosis (MTB) with that of sensitive strains for identifying of unique proteins of these strains and discussing their preliminary application in clinical diagnosis. MethodsProteins of cell wall and membrane of 5 INH-resistant strains and 5 INH-sensitive strains were extracted by density gradient centrifugation.The extracts were subsequently analyzed using weak cation exchange (WCX) liquid chromatography ( LC )followed reverse phase (RP) liquid chromatography to compare the sub-cellular protein patterns. A total of 1280 fractions were collected and identified by matrix assisted laser desorption ionization-time of flight-tandem mass spectrometry (MALDI-TOF MS/MS). The Database for Annotation, Visualization and Integrated Discovery (DAVID) was used for cell component and biological process analysis. Normalized Spectral Abundance Factors (NASF) was used for semi-quantity of protein expression. 5 proteins significantly up-regulated in INH-resistant strains and 2 proteins significantly up-regulated in INH-sensitive strains were selected for ELISA analysis with autologous sera respectively. Results A total of 347 proteins were identified. Cell component analysis showed that 58％ proteins were cells well or membrane proteins. Biological process analysis showed that 31％ proteins involved in carboxylic/monocarboxylic acid biosynthetic and metabolic process, 26％ and 15％ proteins involved in organic acid or fatty acid biosynthetic and metabolic process,while 28％ proteins involved in lipid biosynthetic , metabolic, transport and localization process. O-succinylbenzoate synthase, monooxygenase, hypothetical protein Rv2255c, nicotinate-nucleotide--dimethylbenzimidazole phosphoribosyltransferase and membrane phosphatidate cytidylyltransferase cdsA were up-regulated in INH-resistant strains and fractions contained these proteins could elicit specific antibody response with autologous sera. The A450 was higher than that with INH-sensitive sera. The differences between the INH-resistant sera and the INH-sensitive sera were significant ( t = 0.028, 0.044, 0.066, 0.064, 0.083, all P＜0.01 ). Chain A of Rv2002 Gene Product and Chain A of Crystal Structure Of Rv2632c were up-regulated in INH-sensitive strains and fractions contained these proteins could elicit specific antibody response with autologous sera. The A450 was higher than that with INH-resistant sera. The differences between the INH-sensitive sera and the INH-resistant sera were significant (t=0.053, 0.073, both P＜0.05). ConclusionThe combination of density gradient centrifugation and 2D-LC MS/MS technology is useful in enrichment and identification of differential expressed proteins between INH-resistant and INH-sensitive strains at sub-cellular level. It is useful in finding antigens associated with INH-resistant MTB infection, which may prove useful for further study in the mechanism of INH resistant, as well as interaction between MTB and host.

Objective To investigate technique and clinical effect of total en bloc spondylectomy for thoracic and lumbar chondrosarcoma.Methods From January 2010 to March 2012,6 patients with thoracic or lumbar chondrosarcoma underwent total en bloc spondylectomy.There were 4 males and 2 females,aged from 25 to 54 years (average,38 years).The tumor ranged from T3 to L3; 1 located in T3 and T4,1 in T7,1in T11,1 in L1,1 in L2 and 1 in L3.According to Tomita surgical classification system,there was 1 case of type 2,1 case of type 4,3 cases of type 5 and 1 case of type 6.One patient underwent tumor resection through single posterior approach,while the other 5 patients underwent anterior dissection and posterior resection of tumor.All spines were reconstructed by posterior fixation with pedicle screws and anterior interbody fusion with titanium mesh cages or artificial vertebrae.Results The average amount of blood loss was 3200 ml (range,2100 to 6300 ml).The duration of operation ranged from 3.5 to 12 hours (average,5.5hours).Two patients obtained wide resection,3 obtained marginal resection,and 1 had intralesional margin.The complications included 2 cases of cerebrospinal leak,1 case of pleural effusion and 1 case of pulmonary infection.There was no wound infection and death during peroperative period.All patients were followed up for 6 to 32 months (average,19 months).The neurological function improved from preoperative Frankel C to postoperative Frankel E in 2 cases.All patients obtained bone union 6 to 12 months (average,8 months) after operation.At final follow-up,all patients could walk without aid,and there was no recurrence.Conclusion The total en bloc spondylectomy is an effective method for thoracolumbar chondrosarcoma,which could provide a satisfied tumor control and neurological function improvement.

Aim To study the mechanism of anti-tumor effect of PHⅡ-7 to K562 and K562/A02 cells.Methods The effects of individual and combined doxorubicin on K562 and K562/A02 cells were observed by MTT assay.The coefficient of drug interaction was used to analyse the synergistic effect of PHⅡ-7,obtaining the RNA from the cells stimulated by PHⅡ-7 with different doses to analyse the MDR1 gene expression level.Finally,the cumulation of doxorubicin was observed in K562 and K562/A02 cells after being coped with PHⅡ-7.Results PH Ⅱ-7 had anti-tumor effect with IC50 of (1.37?0.37) ?mol?L-1;(1.48?0.34) ?mol?L-1 for K562 and K562/A02,respectively.It could potentiate the anti-tumor effect of dororubicin with CDI of 0.22 and 0.09 for K562 and K562/A02,respectively.PHⅡ-7 could synergistically inhibit the proliferation of K562 and K562/A02 cells.The decrease of MDR1 expression level depended on the increase of dose of PHⅡ-7 acting on cells.PHⅡ-7 could also develop the cumulation of doxorubicin in cells.Conclusion PHⅡ-7 is not only a Cytotoxinic drug but also can synergistically inhibit the proliferation of K562 and K562/A02 cells with the decrease of MDR1 expression level,especially in K562/A02 cells.

Taxa-4(5),11(12)-diene is the precursor for paclitaxel biosynthesis. The diterpenoid paclitaxel (marketed as Taxol), a plant secondary metabolite isolated from yew, is an effective drug widely used in the treatment of numerous cancers. However, further application of taxol has been restricted due to its low yield in plants and the difficulties in extraction. To increase the intact isoprene flux, we constructed the fusion gene plasmid pBgGGTS and individual cassette plasmid pBgGGgTS to enhance the expression levels of geranylgeranyl diphosphate synthase gene (ggpps) and a taxadiene synthase gene (ts) in Coprinopsis cinerea. These two plasmids were separately transformed into C. cinerea LT2 strain, resulting in several putative transformants. Putative transformants were determined by PCR technique, indicating that 5 out of 13 putative transformants transformed by pBgGGTS and 6 out of 13 putative transformants transformed by pBgGGgTS, respectively. Additionally, the Southern blotting analysis of these 10 transformants confirmed that both ggpps and ts gene were stably integrated into the genome of C. cinerea. Crude extracts from each of the transformants were analyzed. There is no difference in the mycelium extracts among the wild-type LT2 and two types of transformants. However, analysis of culture filtrates indicated that an additional GC peak was found at the retention time of 16.762 min which was absent in the wild type control. The mass fragmentation pattern of this peak had the same diagnostic ions with taxa-4(5),11(12)-diene. According to peak area, the amounts of taxa-4(5),11(12)-diene in each fermented broth were 44 ng/L (transformed with pBgGGgTS) and 30 ng/L (transformed with pBgGGTS), respectively. In conclusion, co-expression of the ggpps and ts gene could increase the taxadiene production in C. cinerea.
Agaricales ; metabolism ; Alkenes ; metabolism ; Diterpenes ; metabolism ; Farnesyltranstransferase ; genetics ; metabolism ; Genetic Engineering ; Isomerases ; genetics ; metabolism ; Paclitaxel ; Plasmids