Isolation and Identification of Eleven Tertiary Alkaloidsin Corydalis decumbens

Eleven tertiarv alkaloids were isolated from Corvdalis decumbens rhizome Their structures were identified as protopine(Ⅶ),allocryptopine(Ⅷ),cryptopine (Ⅸ),muramine(ⅩⅦ);(+)-tetrahydropalmatine(Ⅲ),(+)-kikemanine (Ⅳ),(一)-scoulerine (ⅩⅧ),(十)-bulbo- capnine (ⅩⅨ),and(一)一capnoidine(Ⅰ),(一)一bicuculline(Ⅴ),(+) coriumidine(ⅩⅣ), respecti vely。 Among these,cryptopine,muramine,(+)-kikemanine,(-)-scoulerine,(一)capnoidine,(一)一bicuculline were isolated for the first time from this plant

Effect of New Immunosuppressant-Rapamycin on One-way Mixed Lymphocyte Culture

Objective: Observing human to mouse one-way mixed lymphocyte culture(xMLC) and the effect of new immunosuppressant-Rapamycin on XMLC. Methods: Mouse splenic lymphocyte were collected and treated by mitomycin as activating cell; Human Peripheral blood lymphocytes(hPBL)were separated and gathered as reacting cell; Mouse splenic lymphocyte and hPBL wee mixed to incubate for 1 week; The researchers designed control、RPM groups,and experiment(drugs)grup have different concentration. Results: HPBL in the experiment groups (mixed mouse lymphocyte)proliferated obviously,the amount of3H-TdR in corporation increased evidently(P<0 05,The mean percentage of CD 4, CD 8, LgG, LgM positive cells rose markedly. HPBL in the experiment groups less proliferated,the amount of 3H-TdR incorporation declined,RPM's ic50(50%inhibition concentration)approximately in 1.5 nmol/L; the mean percentage of CD 4, CD 8, IgG, IgM positive cells fell obviously. Conclusion: The human to mouse one-way MLC has obvious lymphocyte proliferation. New immunosuppressants-Rapamycin have powerful effete on XMLC.

Extracapsular invasion as a risk factor for disease recurrence in colorectal cancer

AIM: To evaluate the presence of extracapsular invasion (ECI) in positive nodes as a predictor of disease recurrence disease in colorectal cancer. METHODS: Two hundred and twenty-eight consecutive patients who underwent colorectal resection were identified for inclusion in this study, of which 46 had positive lymph nodes. Among 46 cases with stage Ⅲcolorectal cancer, 16 had ECI at positive nodes and 8 had disease recurrence. The clinical and pathological features of these cases were reviewed. RESULTS: In the univariate analysis, the number of positive lymph nodes and depth of tumor invasion were significantly associated with the presence of ECI at positive nodes. Multivariate analysis demonstrated that only ECI was a predictor of recurrence. The recurrence-free interval differed significantly among patients with ECI at positive nodes.CONCLUSION: Our results suggest that ECI at metastatic nodes can identify which cases are at high risk of short-term disease recurrence in colorectal cancer.

Distribution of Bacteria Injected in Body of Giant Black Shrimp, Penaeus Monodon

Distribution of injected Vibrio anguillarum in body of Penaeus monodon was studied with immunohistochemical method. Bacteria could be detected throughout the experiment in some individuals; however in lymphoid tissue, gill, heart and haemolymph of all vibrio injected shrimp, the bacteria could be observed only 5 min after injection. The bacteria density in haemolymph, haemolymph of the hepatopancreas and gills decreased with time. In the lymphoid organ and heart, the bacteria density was the highest 48 h after injection, then decreased. Nodules could be formed in the heart, lymphoid organ and injection site.

Clinical experiences with 20 cases of single-direction thoracoscopic lobectomy and systematic lymph node dissection for peripheral NSCLC

Objective: To evaluate the safety, efficacy, feasibility of single-direction thoracoscopic lobectomy for peripheral lung cancer. Methods: From December 2009 to March 2011, 20 patients with peripheral lung cancer were treated with single-direction thoracoscopic lobectomy and systemic lymph nodes dissection. Results: Surgeries were successfully performed. No significant complications occurred perioperatively. The average operation time was 193 min, the average blood loss was 234 ml, the average duration of drainage was 6 d, the postoperative hospital stay was 12 d, and the average number of lymph nodes dissected was 16. Conclusion: Single-direction thoracoscopic lobectomy is feasible and safe in the treatment of peripheral lung cancer and can simplify the surgical procedures.

Generation and selection of immunized Fab phage display library against human B cell lymphoma

The approval of using monoclonal antibodies as a targeted therapy in the management of patients with B cell lymphoma has led to new treatment options for this group of patients. Production ofmonoclonal antibodies by the traditional hybridoma technology is costly, and the resulting murine antibodies often have the disadvantage of triggering human anti-mouse antibody （HAMA） response. Therefore recombinant Fab antibodies generated by the phage display technology can be a suitable alternative in managing B cell lymphoma. In this study, we extracted total RNA from spleen cells of BALB/c mice immunized with human B lymphoma cells, and used RT-PCR to amplify cDNAs coding for the κ light chains and Fd fragments of heavy chains. After appropriate restriction digests, these cDNA fragments were successively inserted into the phagemid vector pComb3H-SS to construct an immunized Fab phage display library. The diversity of the constructed library was approximately 1.94× 10^7. Following five rounds of biopanning, soluble Fab antibodies were produced from positive clones identified by ELISA. From eight positive clones, FabC06, FabC21, FabC43 and FabC59 were selected for sequence analysis. At the level of amino acid sequences, the variable heavy domains （VH） and variable light domains （VL） were found to share 88-92% and 89-94% homology with sequences coded by the corresponding murine germline genes respectively. Furthermore, reactivity with membrane proteins of the B cell lymphoma was demonstrated by immunohistochemistry and western blotting. These immunized Fab antibodies may provide a valuable tool for further study of B cell lymphoma and could also contribute to the improvement of disease therapy.

Prevision of Delay in Brazilian Residential Unit Construction Contracts

The aim of this paper is to verify the existence of a clause in contracts of promise of property sale which defines the consequences of delay in the delivery of property, and to discover whether owners demanded any compensation for delays occurred. The number of buildings delivered after the scheduled deadline in two cities between December 2009 and January 2011 was found （n = 542）, and a sample of 82 residential units was taken. In order to obtain the data, a semi-structured questionnaire was given to the homeowners. It was observed that only 35% （n = 29） of the contracts included a penalty clause for delay. However, in 19 of the 29 contracts the clause regarding delay came with other clauses that allowed the company to delay the delivery of the building for up to six months. This clause is considered null in Brazilian legislation. As for compensation, only one （1.82%） owner demanded that the company provide a rental property during the period of delay. It could be concluded that contractors are not observing Brazilian legislation regarding the delay in the delivery of residential units. In addition, conformism is perceived in the attitude of the homeowners.

CORRELATION BETWEEN THE NUMBER OF MITOTIC FIGURES AND THE PERCENTAGE OF Ki67－POSITIVE CELLS IN NON－HODGKIN’S LYMPHOMAS

Proliferative activity in 106 cases of non-Hodgkin’s lymphoma was estimated using the monoclonal antibody Ki67 and by counting the number of mitotic figures. The percentage of Ki67-positive cells was compared with the median number of mitotic figures per square millimeter. Both Ki67 positivity and the number of mitotic figures were found to be greater in high grade than in low grade lymphomas, although there was an overlap between the two grades of malignancy. A close correlation was found between the number of mitotic figures and the percentage of Ki67-positive cells not only in all lymphoma typo taken together (r_s= 0. 834 . P<0. 001). but also in B-cell lymphoma (r_s= 0. 8 1 P< 0. 001) and T-cell lymphoma (r_s = 0. 764. P<0. 00 1) taken seprately. Thus, both methods are useful for the estimation of proliferative activity. but each method has its advantages and disadvantages.

Monoclonal antibodies as therapeutic agents in oncology and antibody gene therapy

Antibodies as therapeutic agents are mostly used in oncology, as illustrated by their applications in lymphoma, breast cancer or colorectal cancer. This review provides a brief historical sketch of the development of monoclonal antibodies for cancer treatment and summarizes the most significant clinical data for the best-established reagents to date. It also discusses strategies to improve the anti-tumor efficacy of antibody therapy, including antibody gene therapy and exploitation of bone marrow derived primary mesenchymal stem cells as the antibody gene transporter.

Loss of CD20 expression in relapsed diffuse large B cell lymphoma after rituximab therapy:a case report and review of the literature

Nowadays, resistance to rituximab has become a major issue in clinical practice. And loss of CD20 may contribute to it. Here we presented a case of loss of CD20 expression in relapsed diffuse large B cell lymphoma treated with rituximab and discuss the incidence, mechanism, influence factors, specific markers, prognosis and treatment of this disease. These results suggested that a post-relapse biopsy after rituximab treatment shguld be performed. CD79a and Pax-5 should be used as the first-line B lineage-specific markers for these patients. Though mechanisms of CD20 decrement are not fully elucidated, the down-regulation of CD20 mRNA is the most probable hypothesis. Recently various new agents are developed, but the prognosis is still poor. Further studies for new treatments are needed.

Investigating the role of^(99m)Tc-TRODAT-1 SPECT imaging in idiopathic Parkinson's disease

Objective:To investigate the role of ^(99m)Tc-TRODAT-1 SPECT in diagnosis and assessing severity of idiopathicParkinson's disease(PD).Methods:Thirty-eight patients with primary,tentative diagnosis of PD and eighteen age-matchednormal controls were studied with ^(99m)Tc-TRODAT-1 SPECT imaging.The regions of interests(ROIs)were drawn manually oncerebellum(CB),occipital cortex(OC)and three transverse plane slice-views of striatums,the semiquantitative BG(back-ground)/[(OC+CB)/2]were then calculated.Results:A lower uptake of ^(99m)Tc-TRODAT-1 in striatums were displayed inthirty-six out of thirty-eight PD patients by visual inspection,compared to controls.In twenty-four PD cases with HYS(Hoehn andYahr scale)stage Ⅰ,a greater loss of DAT uptake was found in striatum and its subregions contralateral striatum to the affectedlimbs than in the same regions of the controls,although the striatal uptake was bilaterally reduced.Using Spearman correlationanalysis showed that the reduction of the uptake ratios significantly correlated with the UPDRS in striatum and all its subregions inthe PD group(P<0.05),a similar change was also found in the putamen by using the rating scale of Hoehn and Yahr (P<0.05).However,analysis of variance(ANOVA)did not show any relationship between the decreasing uptake of ^(99m)Tc-TRODAT-1 andincreasing severity of PD patients,although the specific uptake of ^(99m)Tc-TRODAT-1 was continuously decreased in the striatumby visual inspection with the progress of PD from HYS stage Ⅰ to Ⅲ.Conclusion:^(99m)Tc-TRODAT-1 SPECT imaging may serve asa useful method for improving the correct diagnosis of PD.In assessing the role of ^(99m)Tc-TRODAT-1 SPECT in disease severity ofPD,UPDRS can offer a comprehensive index,although the Hoehn and Yahr assessment may be available in part.

Effects of low dose pre-irradiation on the toxicity of cyclophosphamide

Objective:The aim of the study was to investigate the effects as well as the possible mechanisms of low dose γ-ray pre-irradiation on hepatic damage,DNA damage of peripheral lymphocytes and genetic material damage caused by high dosage of cyclophosphamide(CTX).Methods:Kunming strain male mice were randomly divided into five groups:control group,sham-irradiated group,low dose irradiation group(LDR group),cyclophosphamide chemotherapy group(CTX group) and low dose irradiation combined with chemotherapy group(LDR + CTX group).Having being raised for one week,all the mice were implanted subcutaneously with S180 cells in the left inguen(control group excluded).On days 8 and 11,mice of LDR and LDR + CTX groups were given 75 mGy whole-body γ-irradiation,30 h later mice of CTX and LDR + CTX groups were injected i.p.3.0 mg cyclophosphamide.All the mice were sacrificed on day 13.DNA damage of the peripheral lymphocytes was analyzed using single cell gel electrophoresis(SCGE);ALT activity,total protein(TP) and albumin(ALB) of the plasma were analyzed using automatic biochemistry analyzer;MDA content,SOD and GSH-PX activity of the hepatic homogenate were analyzed using chromometry;genetic material damage was analyzed using micronucleus frequency(MNF) of polychromatoerythrocytes(PCE) in bone marrow.Results:1.Differences of MDA contents,SOD and GSH-PX activity of hepatic homogenate between 5 groups had notable statistical significance(P < 0.01);in control group MDA content was the lowest,SOD and GSH-PX activity were the highest,while in CTX group MDA content was the highest,SOD and GSH-PX activity were the lowest;compared with CTX group MDA content decreased significantly(P < 0.01) and SOD and GSH-PX activity increased significantly(P < 0.05) in LDR + CTX group.2.Differences of ALT activity of plasma between 5 groups had no statistical significance(F = 1.262,P > 0.05).Differences of TP and ALB of plasma between 5 groups had statistical significance(F = 12.879 and 6.336 respectively,P < 0.01);TP and ALB in control group were higher than those of other groups and compared with sham-irradiated group,TP and ALB in LDR group elevated significantly(P < 0.05).3.Differences of DNA damage of peripheral lymphocytes had notable statistical significance(F = 6.383,P < 0.01);DNA damage in control group was the lightest,while DNA damage in CTX group was the severest;compared with CTX group,DNA damage in LDR + CTX group was much lighter(P < 0.05).4.MNF of PCE between 5 groups had remarkable significance(F = 179.652,P < 0.01);compared with control group and sham-irradiated group,MNF in CTX group increased significantly(P < 0.01);compared with CTX group,MNF in LDR + CTX group had a tendency of decline,which had no statistical significance(P > 0.05).Conclusion:1.CTX can damage the hepatic tissue through oxidative stress;75 mGy γ-irradiation before CTX chemotherapy can induce activities of anti-oxidative enzymes,promote elimination of free radicals,so as to alleviate the damaging effects of oxidative stress to hepatic tissue caused by high-dose chemotherapy.2.A 75 mGy γ-irradiation before CTX chemotherapy has no obvious effect on ALT activity of plasma,but may have protective effect on the protein synthesis function of liver.3.High-dose CTX chemotherapy can cause DNA damage of peripheral lymphocytes;75 mGy γ-irradiation before chemotherapy may have certain protective effect on DNA damage.4.CTX has potent mutagenic effect,can cause significant increase of MNF of PCE;75 mGy γ-ray pre-irradiation did not show obvious protection against genetic toxicity of high-dose CTX chemotherapy.

Identification of Pseudomonas syringae pv. lachrymans in Barbados by rep-PCR

Plant disease diagnosis in Barbados is based mainly on traditional biochemical and microbiological cultural techniques with infrequent use of molecular diagnostic techniques. The main objective of this study was to provide a simple, rapid and accurate PCR strategy for identifying plant diseases in Barbados. We used rep-PCR a molecular diagnostic technique which can provide a barcode profile for many bacteria and some pathogenic fungi. Pseudomonas syringe pv. lachrymans was detected in both seed and leaf tissue from an infected field and commercial seed samples of cucumber in Barbados. Pseudomonas syringae pv. lachrymans reference strains were identified by rep-PCR and compared to isolates from field and commercial varieties of cucumber. Leaf and seed isolates had similar molecular profiles to different reference strains of P. syringae pv. lachryrnans and displayed different rep-PCR profiles when compared to each other. Isolates from seed also showed more variation than leaf samples. While this represents the first scientific report of the pathogen on the island, the data suggest that the presence of P. syringae pv. lachrymans in Barbados may be due to both external and internal contamination through the use of imported seed or spread by endemic epiphytes respectively.

Detection of D2-40 monoclonal antibody-labeled lymphatic vessel invasion in esophageal squamous cell carcinoma and its clinicopathologic significance

Objective： This study aims to investigate the clinicopathologic significance of lymphatic vessel invasion （LVI） labeled by D2-40 monoclonal antibody in esophageal squamous cell carcinoma （ESCC）. Methods： Immunohistochemical assay was used to detect the expression of D2-40 and LVI in 107 ESCC patients. Then, the correlation between the clinicopathologic feature and the overall survival time of the patients was analyzed. Results： The lymph node metastasis rates were 70% and 21% in the LVI-positive and LVI-negative groups, respectively. The nodal metastasis rate was higher in the LVI-positive group than in the LVI-negative group. Multivariate regression analysis showed that LVI was related to nodal metastasis （P〈0.001）. The median survival time of the patients was 26 and 43 months in the LVI-positive and LVI-negative groups, respectively. Mthough univariate regression analysis showed significant difference between the two groups （P=0.014）, multivariate regression analysis revealed that LVI was not an independent prognostic factor for overall survival in the ESCC patients （P=0.062）. Lymphatic node metastasis （P=0.031）, clinical stage （P=0.019）, and residual tumor （P=0.026） were the independent prognostic factors. Conclusion： LVI labeled by D2-40 monoclonal antibody is a risk factor predictive of lymph node metastasis in ESCC patients.

Bandwidth-enhanced dual-polarized antenna with improved broadband integrated balun and distributed parasitic element

A bandwidth-enhanced dual-polarized antenna is proposed for 2/3/4/5G applications,which is composed of distributed parasitic elements(DPEs),a main radiator,two improved broadband integrated baluns and a reflector.First,a novel tooth-shape shorted slot line in the improved broadband integrated balun is analyzed to adjust the input impedance of the antenna.Then,DPEs with 2×2 circular plates loading over the main radiator are proposed to improve broadband impedance matching and radiation pattern.By utilizing impedance compensation of the tooth-shaped shorted slot line and the electromagnetic induction of the DPEs,the antenna achieves an enhanced impedance bandwidth and a stable radiation pattern.To verify these ideas,the bandwidth-enhanced dual-polarized antenna was fabricated and measured.The experimental results indicate that the proposed antenna achieves an operating bandwidth of 72.2%(1.69 to 3.60 GHz)with a return loss(RL)less than-15 dB and a port-to-port isolation(ISO)larger than 30 dB.The antenna obtains a half-power beamwidth(HPBW)within(66±5)°and a gain within(9.0±0.6)dBi in the 2/3/4G bands,and an HPBW within(61.5±2.5)°and a gain within(9.8±0.3)dBi in the 5G band.Across the whole band,the cross-polarization discrimination(XPD)and the front-to-back ratio are both larger than 20 dB.

The immunophenotypic changes and clinical effectiveness after treatment of cancer patients with infusion of human peripheral blood lymphocytes stimulated by anti-CD28 and anti-CD80 monoclonal antibodies in combination with radiotherapy and chemotherapy

To investigate the changes on the immunopbenotypes and the clinical effects of treatment of the late cancer patients with infusion of human peripheral blood lymphocytes stimulated by anti-CD28 and anti-CD80 monoclonal antibodies in combination with radiotherapy and chemotherapy, 42 patients with late cancers were collected for study, among which 22 patients were treated with infusion of stimulated lymphocytes in combination with radiotherapy and chemotherapy. The immunological treatment procedure was given twice per week, and one course of treatment consisted of 8 times of giving infusion of lymphocytes. Another 20 patients were selected for control group, in which only radiotherapy and chemotherapy were given without lymphocyte infusions. Flow cytometry was used to examine the immunophenotypes and the clinical symptoms were observed before and after treatments. It was found that the numbers of the CD3^ ＋ , CD4^＋ cells increased, while those of the CD8 ^＋ cells decreased, with an increase of CD4/CD8 radios, but no significant difference existed in case of 22 patients treated with lymphocyte infusion as well as with radiotherapy and chemotherapy. Fifteen patients out of these 22 cases （68.18%）, the immunophenotypes changed obviously with increased numbers of CD3^ ＋ , CD4^ ＋ cells in comparison with those before treatment, and the number of CD95^ ＋ cells was increased after treatment. The PS value in this group of patients decreased after treatment. In comparison with 20 cases in the control group, the immunophenotypes showed no differences before and after treatment. While the PS value decreased obviously. Seven out of the 22 cases （31.83 % ） treated with lymphocyte infusions as well as with radiotherapy and chemotherapy illustrated no major changes in their i mmunophenotypes, compared with the situation before treatment, but the PS value also decreased. In case of treatment with lymphocyte infusions in combination with radiotherapy and chemotherapy, the alteration of phenotypes was reversely correlated with the changes of clinical grades. Although there were 7 cases showing no major alterations of the immunological phenotypes, but their correlation was still evident. In the control group, neither alteration of immunophenotypes nor changes in clinical grades was found. It is concluded that immunotherapy in combination with radiotherapy and chemotherapy can relieve the side effects induced by radiotherapy and chemotherapy and also enhance the therapeutic efforts.

Correlativity of lymph node micrometastases detection in follicular carcinoma of thyroid in different risk groups

Objective:The aim of this study was to explore the relationship between follicular carcinoma of thyroid in different risk groups and lymph node micrometastases.Methods:The keratin-19 of negative neck lymph nodes in 83 cases in routine pathological examination,was detected by SP immunohistochemistry using keratin-19 monoclonal antibody to confirm lymph node micrometastases.All of cases are divided into high risk group,middle risk group and low risk group according to factors related prognosis,the relationship between lymph node micrometastases and different risk groups and follow-up visit documents were analyzed.Results:Fifty-eight neck lymph nodes in 16 cases of 83 cases(19.3%) showed positive lymph node micrometastases,and incidence of lymph node micrometastases was 4/39 in low risk group,5/32 in middle risk group and 7/12 in high risk group,respectively.it showed remarkable difference during 3 groups(P < 0.001).Nine patients in 16 cases with positive lymph node micrometastases showed local recurrence and distant metastases,6 patients in 67 cases with negative lymph node micrometastases showed same result(P < 0.001).Conclusion:Lymph node micrometastases in follicular thyroid carcinoma closely correlated to factors related to prognosis.The detection of lymph node micrometastases can directly assistant postoperative treatment and prognosis evaluation to some extent for follicular thyroid carcinoma.

Stimulation of staphylococcal enterotoxin A combined with PML-RARα peptide on the specifical T-cells against NB4 cell line

Objective： To investigate the effects of staphyococcal enterotoxin A （SEA） on the cytotoxicity of T cells stimulated by PML-RARa peptide in vitro. Methods： Peripheral blood mononuclear cells （MNC） from healthy donor were obtained by density gradient centrifugation on Ficoll-Hypaque, MNC were cultured with PML-RARa peptide and SEA for 20 days. After induction, the cytotoxicity of T cells induced against NB4 and K562 cell lines were examined by Cell Counting Kit-8 （CCK-8）. The CD4 and CD8 surface markers on the harvested CD3^＋ T cells were detected by flow cytometry （FCM）. Results： The cytotoxicity of T cells induced by PML-RARa peptide with SEA was higher than that of T cells induced only by PML-RARa peptide against NB4 cells. The FCM assay showed that the ratio of CD4^＋/CD8^＋ T cells were gradually decreased in both groups of PML-RARα peptide whether with SEA or not at the intervals of day 5,10 and 20 day after induction, but the most significantly decreased by PML-RARe peptide with SEA. Conclusion： The specific cytotoxicity of T cells induced by PML-RARa peptide against NB4 cells could be enhanced with superantigen SEA.

Listeria monocytogenes in Live Mytillis galloprovincialis Collected from Butrinti Lagoon Located in South Part of Albania

In 2009, 201 samples of live bivalve mussels （Mytillus galloprovincialis） were analyzed for presence of Listeria monocytogenes. The samples were collected from three sampling points （BM l-south, BM l-north and BMl-west） in Butrinti lagoon located in the south of Albania. The examination confirmed incidence of this pathogen with average value 4.5%. Based on the results, high incidence was reported in winter with 6.7% of analyzed samples. The incidence of L. monocytogenes was also reported respectively in spring of 2009 with 4.35% and in autumn with 2.3% of controlled samples. There is no big difference in values of incidence of L. monocytogenes between summer and autumn. The northern part of the lagoon was more indicated by contamination of L. monocytogenes. Examination of 67 samples of M. galloprovincialis, collected in the northern part of this lagoon, showed five cases or about 7.5% of them to be confirmed as positive. Referring to the study results, periodic controls of the sea products, particularly of live bivalve mollusks, for presence ofL. monocytogenes remains a necessary objective of food inspection in Albania.