Complex ecological and socioeconomic impacts on medicinal plant diversity.

Medicinal plant diversity (MPD) is an important component of plant diversity. Over-collection based on medicinal and economic value has the potential to damage the stability of the regional ecosystem. It is important to understand the current distribution of MPD and the factors influencing it. However, it is still unclear whether environmental and socioeconomic conditions have an impact on their distribution. We selected the Inner Mongolia as a representative study area which covers a wide area, accounting for 12.29% of China's national land area and 0.79% of the world's land area. At the same time, the region is a long-standing traditional medicinal area for Mongolians in China. Therefore, the region is significantly influenced by changes in environmental factors and socio-economic factors. We used 9-years field survey of the distribution of medicinal plants in Inner Mongolia for assessing the distribution of MPD as influenced by environmental and socioeconomic activities by combining spatial analyses, species distribution models, and generalized additive models. The results from the spatial analysis show that the western region of Inner Mongolia is the main cold spot area of the MPD, and the central-eastern and northeastern regions of Inner Mongolia are the main hot spot areas of the MPD. At the same time, the distribution of cold spots and hot spots of MPD is more obvious at large spatial scales, and with the refinement of spatial scales, the cold spots in scattered areas are gradually revealed, which is indicative for the conservation and development of MPD at different spatial scales. Under the future climate change of shared socioeconomic pathways (SSP), areas with high habitat suitability for medicinal plants remain mainly dominated by the Yellow River, Yin Mountains, and Greater Khingan Range. Notably, the SSP245 development pathway remains the most significant concern in either long- or short-term development. The nonlinear relationship between the driving factors of MPD at different spatial scales shows that temperature, precipitation and socioeconomic development do have complex effects on MPD. The presence of a certain temperature, altitude, and precipitation range has an optimal facilitation effect on MPD, rather than a single facilitation effect. This complex nonlinear correlation provides a reference for further studies on plant diversity and sustainable development and management. In this study, the spatial distribution of medicinal plant resources and the extent to which they are driven by ecological and socioeconomic factors were analyzed through a macroscopic approach. This provides a reference for larger-scale studies on the environmental and socioeconomic influences on the distribution of plant resources.

[Application of remote sensing technology in medicinal plant resources].

The sustainable use of medicinal plants is the foundation of the inheritance of traditional Chinese medicine(TCM) and the acquisition of information on medicinal plants is the basis for the development of TCM. The traditional methods of investigating medicinal plant resources are disadvantageous in strong subjectivity and poor timeliness, making it difficult to real-time monitor medicinal plant resources. In recent years, remote sensing technology has become an important means of obtaining information on medicinal plants. The application of this technology has made up for the shortcomings of traditional methods. The open-access remote sensing data with medium spatial resolution satellites provide an opportunity for extracting information on medicinal plant resources. This study firstly introduced the principles of remote sensing technology, summarized the satellites and the parameters commonly used in the field of medicinal plant resources, and compared the survey methods of remote sensing technology with traditional methods. Secondly, it reviewed the applications of remote sensing technology in the extraction of information on the cultivation of medicinal plants and the common methods for extracting the planting structure information of medicinal plants based on remote sensing technology. Thirdly, the applications of remote sensing technology in the investigation and monitoring of medicinal plants were further analyzed with the research objects divided into wild and cultivated medicinal plants according to the characteristics of the habitats. Finally, it pointed out the key unsolved technical problems in the remote sensing monitoring of medicinal plant resources, and proposed solutions for the intelligent information processing of medicinal plants based on remote sensing big data, which is expected to provide references for the development of remote sensing technology in derivative application in medicinal plant resources.

[Research of estimation methods on medicinal plant resources reserves].

The medicinal plant resource reserve refers to the natural resources of medicinal plants in a certain time and a certain region within the scope of the volume. In recent years, with the demand of medicinal plant resources surging and the change of the environment and human intervention factors, the medicinal plant resources reserve had accelerated pace of change. It is the prerequisite and basis for the development and utilization of medical plants that how to quickly and accurately attain reserve of some medicinal plants resources, the selection of suitable and accurate estimating method is reliable basis and can guarantee medicinal plant reserve survey, and also is one of the key reserve investigation of success. This paper systematically summarized the estimation method of medicinal plants in recent 30 years, and discussed the basic principle, the estimation model of development and evolution, advantages and disadvantages and applicability, and it aimed to improve the accuracy about reserves survey of medicinal plant resources, and provide scientific and reliable support data to medicinal plants resources for sustainable development and utilization of resources.

Autoantibodies in Chinese patients with chronic hepatitis B: prevalence and clinical associations.

AIM: To investigate the prevalence of autoantibodies and their associations with clinical features in Chinese patients with chronic hepatitis B (CHB). METHODS: A total of 325 Chinese patients with CHB were enrolled in this retrospective, hospital-based study. Patients with chronic hepatitis C (CHC), autoimmune hepatitis (AIH), or primary biliary cirrhosis (PBC) were included, with healthy donors acting as controls. A panel of autoantibodies that serologically define AIH and PBC was tested by indirect immunofluorescence assay and line immunoassay. The AIH-related autoantibody profile included homogeneous anti-nuclear antibodies (ANA-H), smooth-muscle antibodies, anti-liver kidney microsome type 1, anti-liver cytosolic antigen type 1, and anti-soluble liver antigen/liver pancreas; the PBC-related antibodies were characterized by ANA-nuclear dots/membranous rim-like, anti-mitochondrial antibodies-M2 (AMA-M2), anti-BPO (recombinant antigen targeted by AMA-M2), anti-Sp100, anti-promyelocytic leukemia protein (anti-PML), and anti-gp210. The dichotomization of clustering was used to unequivocally designate the AIH or PBC profiles for each case. Anti-Ro52 antibodies were also tested. RESULTS: The prevalence of any autoantibody in CHB amounted to 58.2%, which was similar to the 66.2% prevalence in CHC, significantly higher than the 6.7% in the healthy controls (P < 0.001), and lower than the 100% found in AIH and PBC (P = 0.004 and P < 0.001, respectively). There were more anti-PML and anti-gp210 antibodies among the CHB patients than the CHC patients (11.1% vs 0%, P = 0.003; 12.6% vs 0%, P < 0.001, respectively). The prevalence and titer of AMA, anti-BPO, anti-PML, and anti-gp210 were higher in PBC than in those with CHB. Among the CHB patients, the prevalence of ANA, especially ANA-H, was significantly lower in patients with compensated and decompensated cirrhosis compared with patients without cirrhosis. Thirty-eight cases of hepatocellular carcinoma (HCC) in CHB showed a significant difference compared with non-HCC patients in the prevalence of anti-PML (0% vs 12.5%, P = 0.013). Dichotomization of the autoantibodies revealed that the PBC profile was more prevalent in patients with CHB than in those with CHC, and that it was strongly correlated with both compensated and decompensated cirrhosis. In contrast, the prevalence of the AIH profile was significantly higher in non-cirrhosis patients with CHB than in those with compensated cirrhosis (18.5% vs 8.2%, P = 0.039). Moreover, the AIH profile was also closely associated with hepatitis B e-antigen positivity. CONCLUSION: ANA-H could be an indicator of early-stage CHB. Dichotomizing the autoantibody profiles revealed that the PBC profile is strongly associated with cirrhosis in CHB.

[Cloning and expressing of tissue inhibitor of metalloproteinases I gene fragment and preparation of monoclonal antibodies against the recombinant protein].

OBJECTIVE: To prepare the monoclonal antibody (mAb) against tissue inhibitor of metalloproteinases I (TIMP-I) fusion protein. METHODS: TIMP-I gene was amplified from fibrotic human liver tissue by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-TIMP-I and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot were used to detect specificity of mAbs. RESULTS: The prokaryotic plasmid expressing the recombinant protein was constructed, and the TIMP-I recombinant protein was expressed and purified. Four hybridoma cell lines that secreted anti-TIMP-I mAbs were obtained. 3 of 4 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with TIMP-I protein. CONCLUSION: The TIMP-I recombinant protein is highly purified and has strong antigenicity. The anti- TIMP-I mAbs were prepared successfully.

[Establishment of a purificatory method for alpha-fetoprotein variant by affinity adsorption].

OBJECTIVE: To establish a purificatory method of alpha-fetoprotein variant (AFP-L3) based on microspincolumn with lens culinaris agglutinin (LCA). METHODS: LCA was isolated by ammonium sulfate precipitation method from lens culinaris. AFP-L3 affinity adsorption microspincolumns which were made from LCA coupled with activated Sepharose 4B were prepared. By adding into the centrifuge column, serum was absorbed and eluted to purify AFP-L3. The results of purified AFP-L3 detection of 10 cases AFP positive sera by electro-chemiluminescence immunoassay were compared with traditional crossed affinity immunoelectrophoresis. RESULTS: 8 of 10 cases AFP-L3 concentration were greater than 5 ng/ml in purified sera. Six cases show positive reaction in affinity immune cross electrophoresis experiment. CONCLUSION: Successfully established purification method of AFP-L3 by affinity absorption based on microspincolumn. The method was more conducive to clinical laboratory applications due to its high sensitive and easy operation.

[The investigantion of comparative experiments for different clinical laboratory instruments].

OBJECTIVE: To estimate the consistency of two VITROS 3600 chemiluminescent analyzers according to the requirement of ISO15189. METHODS: Verification tests were made for precision and accuracy of anti-HCV in two instruments. While 40 serum samples including Anti-HCV negative (10 cases) , positive (10 cases) , and weakly positive (20 cases). and the test results were statistical analised. RESULTS: Two instruments negative and positive control samples intra-batch precision and coefficients of variation were 5% , 4% and 7. 14% , 7. 23% , inter-batch precision and coefficients of variation were 9. 47% , 7. 7% and 8.04%, 7. 6%, are less than requirement CV (15%) by ISO15189. The accuracy of two instrument were 100% , The test results of the control samples showed no significant difference (P < 0. 05). The correlation analysis of the test results of clinical samples R2 =0. 9984, with good consistency. CONCLUSION: Test results of two Vitros 3600 has good consistency and comparability.

[Establishment of a detection method for hepatitis B virus large surface protein (HBV-lP)].

OBJECTIVE: To establish enzyme-linked immunosorbent assay (ELISA) for detection of hepatitis B virus large surface protein(HBV-LP) in serum. METHODS: A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of HBV-LP as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as sensitivity, specificity, stability and so on. RESULTS: The detection limit was 5 ng/ml. Interassay and intra-assay RSD were both less than 10%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%. CONCLUSION: Established ELISA for determination of serum HBV-LP has high sensitivity and repeatability. Enzyme-linked immunosorbent assay;

[Microplate chemiluminescence enzyme immunoassay for the quantitative analysis of tissue inhibitor of metalloproteinases I].

OBJECTIVE: To establish microplate chemiluminescence enzyme immunoassay (CLEIA) for quantitative analysis of tissue inhibitor of metalloproteinases I (TIMP I) in human serum. METHODS: A sandwich reaction was preformed with horseradish peroxidase(HRP) labeled monoclonal antibody of TIMP I as the catalytic enzyme and the H2O2-luminol as the luminescence reagent. Several physical and chemical parameters were studied and optimized such as immunoreaction conditions, the dilution ratio of TIMP I-HRP, luminescence reaction time and so on. In order to evaluate the method, recovery test, heat stabilization test and comparison test were carried out. RESULTS: The linear range was 0. 2-12 ng/ml with r = 0.996. The detection limit was 0.12 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 100.6%, 96.5% and 106.5%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0. 998 and RSD lower than 6%. The detected results with CLEIA closely corresponded to those with imported ELISA in 60 patients sera with liver fibrosis. CONCLUSION: Established CLEIA for quantity determination of serum TIMP I has high accuracy, sensitivity and repeatability.

[Cloning and expressing of golgi protein73 gene fragment and preparation of monoclonal antibodies against the recombinant protein].

OBJECTIVE: To clone and express human Golgi glycoprotein73 protein, and prepare the monoclonal antibody (mAb) against the protein. METHODS: GP73 gene was amplified from HepG2 cells by RT-PCR, then ligated with pQE31 to form recombinant plasmid pQE-GP73 and transformed into E. coli BL21. The protein induced by IPTG was purified by 6 x His-tag and used to immunize the BALB/c mice. The specific monoclonal antibodies (mAbs) were prepared by the cell fusion technique. Western Blot was used to detect specificity of mAbs. RESULTS: The prokaryotic plasmid expressing the recombinant protein was constructed, and the GP73 recombinant protein was expressed and purified. Five hybridoma cell lines that secreted anti-GP73 mAbs were obtained. 2 of 5 mAbs were the IgG1 subtype. Western Blot indicated the mAbs showed specific combination with GP73 protein. CONCLUSION: The GP73 recombinant protein is highly purified and has strong antigenicity. The anti-GP73 mAbs were prepared successfully.

[Establishment of a quantitative detection method for Golgi protein73].

OBJECTIVE: To establish enzyme-linked immunosorbent assay (ELISA) for quantitative detection of Golgi protein73 (GP73) in serum. METHODS: A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of GP73 as the catalytic enzyme. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on. RESULTS: The linear range was 25-500 ng/ml. The detection limit was 18.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 95.3%, 92.6% and 103.7%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.98 and RSD lower than 10%. CONCLUSION: Established ELISA for quantity determination of serum GP73 has high accuracy, sensitivity and repeatability.

[Quantitative detection of serum procollagen III N-terminal peptide chemiluminescence enzyme immunoassay].

OBJECTIVE: To establish chemiluminescence enzyme immunoassay (CLEIA) for quantitative detection of procollagen III N-terminal peptide (P III NP) in serum. METHODS: A sandwich reaction was preformed with horseradish peroxidase labeled monoclonal antibody of P III NP as the catalytic enzyme and the luminol as the luminescence reagent. Several reactions liquid's concentration and reaction conditions were optimized. The method was evaluated in all aspects such as linear range, sensitivity, specificity, stability and so on. The CLEIA was compared with imported ELISA kits, by detecting clinical serum. RESULTS: The linear range was 0.8-85 ng/ml. The detection limit was 0.5 ng/ml. Inter-assay and intra-assay RSD were both less than 10%. The recoveries of three different spiked concentration samples were 96.2%, 91.2% and 101.1%. After stored at 4 degrees C and 37 degrees C for 3, 5, 7 days, the analysis showed correlation coefficient higher than 0.99 and RSD lower than 6%. The detected results of clinical sera with CLEIA closely corresponded to those with imported ELISA. CONCLUSION: Established CLEIA for quantity determination of serum P III NP has high accuracy, sensitivity and repeatability.

[Establishment of confirmatory test for suspicious hepatitis B surface antigen positive samples].

OBJECTIVE: Establish a confirmatory test based on ELISA, and use to verify the authenticity of HBsAg weak positive samples, pick and get rid of the false result, and avoid the mistake diagnosis. METHOD: The particles (reagent A) coated by streptavidin and biotinylated HBsAb (reagent B) were mixed in different proportions, then neutralized with serum whose the COI of HBsAg > 20 by ELISA in order to identify the activity of HBsAb in confirmatory reagent. 30 pieces of HBsAg weak positive serum neutralized with the confirmatory reagent, the serum were considered to be positive if rate of decline of HBsAg COI > 50%. The results were compared to Roche confirmatory Kit. RESULT: Confirmatory reagent was able to neutralized with HBsAg. 24 of 30 pieces of HBsAg weak positive samples were judged to be positive, while 6 poeces were negative. The ELISA comfirm method is fully consistent with Roche confirmatory Kit. CONCLUSION: The ELISA confirmatory test for suspicious HBsAg positive samples is a simple, accurate and low cost initial validation method, After further clinical trials, should be widely applied.

[Comparison of two different methods to detect HIV antibodies].

OBJECTIVE: Evaluated the chemiluminescence and enzyme-linked immunosorbent assay (ELISA) to detect HIV antibodies, and compared the results, to provide a reference for the selection and clinical application of HIV screening. METHODS: 3000 cases of our hospital patients were measured by enzyme-linked immunosorbent assay and chemiluminescence immunoassay, using comfirmming experimental results as gold standards. Comparing sensitivity, specificity and other Indicators. RESULTS: In the diagnosis of HIV infection, enzyme-linked immunosorbent assay and chemiluminescence immunoassay had no significant difference. The positive rate of enzyme-linked immunosorbent assay was 0.93%, while the sensitivity and specificity were 89.66%, 99.93%, the positive rate of chemiluminescence immunoassay was 1.03%, while the sensitivity and specificity were 100%, 99.93%, respectively. CONCLUSION: Both methods are suitable for screening of HIV, having high specificity, and chemiluminescence has greater sensitivity than ELISA.

[Prokaryotic expression, purification of human papillomavirus type 16 E1 protein and preparation of it's antiserum].

AIM: To express and purify the human papillomavirus type 16 E1 protein in E.coli and prepare the antibody against HPV-16 E1. METHODS: HPV-16 E1 gene was amplified by PCR and cloned into prokaryotic expression vector pMAL-p2x, and the recombinant plasmid was transformed into E.coil BL-21. We optimize the soluble expression condition of fusion protein by induction with different IPTG concentration and different temperature. The expressed fusion protein was purified by mahose affinity column Chromatography. To prepare the anti-serum, New Zealand white rabbits were immunized with purified HPV-16 E1 protein via hypodermic and volar. Western blot and ELISA analyzed the serum's specificity against HPV-16 E1 and serum titers. RESULTS: Restriction endonuclease analysis and DNA sequencing showed HPV-16 E1 was cloned into the plasmid pMAL-p2x. Based on the optimization experiments, it concluded that the best soluble expression conditions for the HPV-16 E1 fusion protein involved addition of IPTG to a final concentration of 0.5 mmol/L and then further incubation at 28°C. The purity of the HPV-16 E1 fusion protein was over 95.7% after purification. ELISA and Western blotting showed the titers of the anti-serum were above 1:640 000, and the anti-serum can specifically bind with HPV-16 E1 protein. CONCLUSION: We have ingathered the HPV-16 E1 fusion protein by expressing in E.coli and purifying, and the antibody against HPV-16 E1 was prepared with the fusion protein immunizing New Zealand white rabbits. This work will provide an antigen and detection antibody for further study on the HPV-16 E1 function.

[Establishment of confirmatory test for HBsAb in serum of coexistence of hepatitis B surface antigen and antibodies to HBsAg].

OBJECTIVE: Establish a kind of confirmation method based on ELISA, and use to verify authenticity of HBsAb + in HBsAg + HBsAb + serum, pick and get rid of the false masculine gender the result, and avoid the mistake diagnosis. METHOD: Collect 60 pieces of serum whose thick degree of HBsAg at 1000 COI above tested by ECLIA as confirm serum, mixed the confirm serum of different dilution with HBsAb positive serum to screen and verify best thick degree of HBsAg. Collected 40 pieces of HBsAg + HBsAb + serum, ELISA tested the descend rate of HBsAb COI after neutralized with confirm serum in order to confirm authenticity of HBsAb + in pieces of HBsAg + HBsAb + serum. RESULT: When thick degree of HBsAg is 2000 COI, the performance of neutralization to HBsAb is best. The ELISA confirmatory test is fully consistent with the ECLIA method with true positive of 37 pieces of HBsAg + HBsAb + serum while false-positive of 3 pieces of serum. CONCLUSION: The ELISA confirm method is a simple, accurate and low cost initial validation method.

[Evaluation of the Helicobacter pylori stool antigen test].

OBJECTIVE: To evaluate the clinical value of an enzyme-linked immunosorbent assay by the Helicobacter pylori stool antigen (HpSA) test for the detection of H. pylori infection. METHODS: 328 patients were measured upper gastrointestinal endoscopic examination which as gold standards and HpSA test in the meantime, comparing accuracy, sensitivity, specificity and other Indicators. RESULTS: In the diagnosis of Hp infection, HpSA test had no significant difference comparing with gold standards and had a P value of over 0.5. The sensitivity of HpSA test was 94.6%, while the specificity, veracity, expected positive value, and expected negative value were 96.9%, 96.3%, 89.7% and 98.4%, respectively. CONCLUSION: The H. pylori stool antigen test is a simple, non-invasive method for accurate diagnosis of H. pylori infection.

[Evaluation on clinical value of serum CA-125 level in hepatitis cirrhosis].

OBJECTIVE: To determine the association between elevated levels of serum cancer antigen (CA) 125 and hepatitis cirrhosis in different stage, and also to explore the clinical application value of serum CA-125. METHODS: During June to December in 2008, 200 cases with hepatitis cirrhosis were random selected in our hospital. CA-125 levels were measured by electrochemiluminescence immunization assay in sera collected from these cases which were termed with Child-Paugh classification and analyzed by SAS. RESULTS: Serum CA-125 levels were correlated closely with ascites, primary peritonitis and liver function Child-Paugh classification, but no associated with primary carcinoma of liver and other liver function index,such as ALT, AST, ALB, TBIL and PT. CONCLUSION: The levels of serum CA-125 in hepatitis cirrhosis patients were osculating correlating with lesion of liver and ascite degree, could serve as a sensitive and conventional laboratory index for liver lesion degree and monitoring ascite generation. It was necessary to further study on the association with serum CA-125 level with primary hepatic carcinoma.

[Evaluation of the ELISA and the enhanced chemiluminescence immunoassay in use to determine the serum markers for liver fibrosis].

OBJECTIVE: To evaluate the detection method of ELISA and Enhanced Chemiluminescence Immunoassay (ECLIA) in use to determine serum hyaluronate acid (HA), laminin (LN), type IV collagen (IV-C) and type III procollagen (PC III). METHODS: 253 patients with chronic hepatitis B were determined the four liver fibrosis serum markers with both the ECLIA and ELISA, and then compared with pathology results separately. RESULTS: Both the detection results of ELISA and ECLIA can reflect that the patient's liver fibrosis from hepatitis to liver cirrhosis aggravated gradually. Compared with ELISA, the results of ECLIA and pathology have a better correlation. CONCLUSIONS: The detection of four liver fibrosis serum markers by ECLIA could indicate the better the response of the state of live fibrosis.

[The studies on early warning of Lens culinaris-reactive alpha-Fetoprotein (AFP-L3) detecting in primary hepatocellular carcinoma].

OBJECTIVE: To explore the significance of Lens culinaris-reactive alpha-Fetoprotein (AFP-L3) detection in primary hepatocellular carcinoma. METHODS: AFP-L3 was isolated by using microspin column coupled with lens culinaris agglutinin (LCA), AFP and AFP-L3 were detected with chemiluminescent immunoassay, the proportion of AFP L3 levels were calculated, and the relationship between the elevated AFP-L3 (%) levels and benign and malignant liver disease was analyzed. RESULTS: There were significant differences in positive rate between the patients of HCC, suspected HCC and other liver disease (81.80%, 73.68%, 11.80%, respectively, P < 0.05). Among the undetermined HCC (suspected HCC, liver disease) patients, 12 out of 21 cases of AFP-13 positive were diagnosed to be HCC within 6 months, and 6 of them were diagnosed to be the single small HCC at the early stage through B-Ultrasonic Diagnosis or CT. Among 62 cases of AFP-L3 negative, 3 cases were diagnosed to be HCC within 6 months and the risk of occurrence of HCC for AFP-L3 positive increased 11.9 times. CONCLUSION: AFP-L3 has no correlation with AFP value, and it can be used as an independent HCC diagnosis factor. The detection of AFP-L3 has a significant implication for the identification of benign or malignant liver disease and the early stage predictive diagnosis of HCC while AFP increases.