The dynamic of endophytic bacteria at different growth stage of tomato and use of these endophytic bacteria to control tomato bacterial wilt were studied. The results showed that endophytic bacteria could be found in the tomato seeds and their quantities reached the highest peak in the adult plants both in resistant and susceptible cultivars. The amount of endophytic bacteria in adult plants of resistant tomato cultivars was 2.43?10~5CFU/g FW in the root and 22.9?10~4 CFU/g FW in the stem, while the amount of endophytic bacteria in adult plants of susceptible tomato cultivars was 9.8?10~4CFU/g FW in the root and 13.4?10~4CFU/g FW in the stem respectively. Seventeen strains of endophytic bacteria from resistant cultivars and only seven strains from susceptible cultivars were found to be antagonistic to Ralstonia solanacearum. In addition, some strains of endophytic bacteria had the abilities of promoting tomato seed germination and controlling tomato bacterial wilt, among which, strain 5R and 3R had better control effect of 91.7% and 81.3% respectively.

Objective To establish a method of detecting hepatitis B virus x antigen (HBxAg) and antibody to HBxAg (anti-HBx) and to demonstrate its clinical significance of HBxAg and anti-HBx in sera from patients of chronic hepatitis B (CHB),liver cirrhosis (LC) and hepatocellular carcinoma (HCC). Methods Full length HBx gene was cloned into pET30a(+),a prokaryotic expression vector,named pET30a-X.It was transformed into Escherichia coli BL21 (DE3),followed the fusion protein of HBx-His was induced by IPTG.The purified fusion protein was used to immunize rabbit as an antigen to generate polyclonal antibody to HBx protein.The method of enzyme-linked immunosorbent assay (ELISA) was established by using purified fusion protein and generated antibody,which was used to detect HBxAg and anti-HBx in sera from patients of CHB,LC,HCC and normal healthy people.Results The positive rates of HBxAg/anti-HBx were 8.7%/10.4% for CHB,17.9%/40.6% for LC,and 9.8%/34.4% for HCC, respectively.In statistics,the positive rates of anti-HBx in LC and HCC were higher than that in CHB (P

AIMTo study the excretion of (-)-clausenamide in rats.

METHODSThe urine, feces and bile were collected at predetermined time points after (-)-clausenamide was orally administrated to 6 rats (30 mg x kg(-1)). The concentrations of (-)-clausenamide and its metabolite 6-OH-(-)-clausnamide were determined by HPLC-MS/MS method using glipzide as the internal reference, and the accumulative excretion amount of (-)-clausenamide and 6-OH-(-)-clausenamide was calculated in the urine, feces and bile, separately.

RESULTS(-)-Clausenamide was recovered mostly (44%) from feces in 112 hours, 7.1% was found from urine in 120 hours and 0.013% was detected from bile in 24 hours. The accumulative excretions of 6-OH-(-)-clausenamide were 0.92% , 0.46% and 0.0003% of the administered dose from feces, urine and bile, respectively.

CONCLUSIONThe major amount of (-)-clausenamide was recovered from feces after (-)-clausenamide was orally administrated to rats (30 mg kg(-1)).

Administration, Oral ; Animals ; Bile ; metabolism ; Chromatography, High Pressure Liquid ; Clausena ; chemistry ; Feces ; chemistry ; Female ; Lactams ; chemistry ; pharmacokinetics ; urine ; Lignans ; chemistry ; pharmacokinetics ; urine ; Male ; Mass Spectrometry ; Neuroprotective Agents ; administration & dosage ; pharmacokinetics ; urine ; Plant Leaves ; chemistry ; Plants, Medicinal ; chemistry ; Rats ; Rats, Sprague-Dawley ; Stereoisomerism

OBJECTIVETo evaluate the effects of the Chinese herbal formula Wuzi Yanzong Pill (, WYP) on the spermatogenesis and specific secretory functions of Sertoli cells in rat model and to investigate the underlying mechanism.

METHODSFive groups of male Sprague-Dawley rats including the control group, the model group, the low-dose WYP group, the medium-dose WYP group and the high-dose WYP group (5 in each group) were treated daily with vehicle, multiglycosides of Tripterygium wilfordii Hook f (GTW) either alone (20 mg/kg) or followed by WYP (0.5, 1.0, or 2.0 g/kg daily), respectively for 30 days. Serum levels of follicle-stimulating hormone (FSH), inhibin B (INHB) and testosterone (T) were evaluated using enzyme-linked immunosorbent assay. Androgen-binding protein (ABP) gene expression and transferrin (TF) protein expression in testis tissue specimens of all rats were determined using real-time reverse transcriptase polymerase chain reaction and Western blotting analysis, respectively. Histopathological alterations in the testis were determined using Johnsen's score.

RESULTSThe toxicity of GTW towards Sertoli cell secretory functions and spermatogenesis was accompanied by increased serum FSH concentrations and decreased INHB and T concentrations. Upregulated ABP mRNA levels, and decreased TF protein expression and Johnsen's scores were detected in the model group compared with the control group P<0.05 or P<0.01). Oral high-dose WYP administrations to GTW-treated rats effectively alleviated all of the GTW-induced changes in specific secretory functions of Sertoli cells (ABP, INHB and TF). Furthermore, serum T level and Johnsen's score of the testis increased greatly compared with the model group (P<0.01).

CONCLUSIONWYP has the ability to improve the spermatogenesis, possibly through modulating the secretory proteins expression of Sertoli cells.

Androgen-Binding Protein ; genetics ; metabolism ; Animals ; Blotting, Western ; Drugs, Chinese Herbal ; pharmacology ; Follicle Stimulating Hormone ; blood ; Gene Expression Regulation ; drug effects ; Inhibins ; blood ; Male ; RNA, Messenger ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley ; Sertoli Cells ; drug effects ; secretion ; Spermatogenesis ; drug effects ; Tablets ; Testis ; cytology ; metabolism ; Testosterone ; blood ; Transferrin ; metabolism

AIMTo study the four diastereomers of leucogen and structure of the related substances.

METHODSLC-DAD, LC-MS/MS and LC- 1H NMR were used. LC was carried out with a Phenomnex Luna C18 (250 mm x 4.60 mm ID, 5 microm) column and a mobile phase of water-acetonitrile-glacial acetic acid (58:42: 0.3).

RESULTSThe structures of leucogen and its related substances were identified. Leucogen and the related substances were found to have four diastereomers in solution state separately. The stability and transformation of the four diastereomers were analyzed and 3R4S5R was found to be more stable than the others according to quantum calculations.

CONCLUSIONLeucogen have four diastereomers in solution state and it can transform from one diastereomer to the others, and the 3R4S5R is more stable than the others.

Chromatography, Liquid ; Magnetic Resonance Spectroscopy ; Molecular Structure ; Quantum Theory ; Solutions ; Spectrometry, Mass, Electrospray Ionization ; Stereoisomerism ; Thiazolidines ; analysis ; chemistry

OBJECTIVETo study the loss of heterozygosity (LOH) on chromosome 3p in thyroid tumors.

METHODSLOH at 11 microsatellite loci was analyzed in 74 cases of thyroid tumors (including 20 follicular adenomas, 24 follicular thyroid carcinomas and 30 papillary thyroid carcinomas) by polymerase chain reaction and silver stain.

RESULTSLOH on chromosome 3p was detected in 71% of follicular thyroid carcinoma (17/24), 30% of the papillary thyroid carcinoma (9/30) and 10% of the follicular adenoma (2/20) case. Two minimal common deleted regions (CDR) (3p26-pter and 3p14.2-3p22) involving significant sites of LOH has identified in follicular thyroid carcinoma. There was also one CDR (3p25. 2-26.1) in papillary thyroid carcinoma.

CONCLUSIONSLOH is more frequently identified in follicular thyroid carcinoma than in papillary thyroid carcinoma and follicular adenoma. The 3 CDR on chromosome 3p may harbor tumor suppressor genes involved in the pathogenesis of follicular thyroid carcinoma and papillary thyroid carcinoma.

Adenocarcinoma, Follicular ; genetics ; Adenoma ; genetics ; Adult ; Aged ; Carcinoma, Papillary ; genetics ; Chromosome Mapping ; Chromosomes ; Chromosomes, Human, Pair 3 ; genetics ; Female ; Genes, Tumor Suppressor ; physiology ; Heterozygote ; Humans ; Loss of Heterozygosity ; Male ; Microsatellite Repeats ; Middle Aged ; Thyroid Neoplasms ; genetics ; Young Adult

This study was designed to compare the curative effect, prognosis and safety of different preconditioning regimens for patients who received autologous hematopoietic stem cell transplantation (AHSCT) for malignant lymphoma (ML). The clinical data of 100 ML patients (Sep 1992 to Aug 2010 in 307 Hospital) were retrospectively analyzed, and were divided into two groups by different preconditioning regimens: the high-dose chemotherapy preconditioning group and high-dose chemotherapy/radiotherapy preconditioning group. The overall survival (OS) rate, progress free survival (PFS) rate and adverse effect were analyzed. The results showed that until Feb 2011, the median follow-up was 33.5 months. All patients were engrafted and their hematopoiesis was reconstituted. The median time of WBC recovery up to > 1.0×1.0(9)/L in high-dose chemotherapy preconditioning group and high-dose chemotherapy/radiotherapy preconditioning group were (6.0 ± 0.4) d and (8.2 ± 0.4) d, platelet up to > 20.0×1.0(9)/L in two groups were (7.1 ± 0.8) d and (11.4 ± 2.5) d (P < 0.05). The 3-year OS rate of the two groups were 67.3% and 68.9%. 5-year OS rates of two groups were 62.8% and 60.6%, 10-year OS rates of two groups were 57.6% and 56.2% respectively; 3-year PFS of two group were 63.6% and 63.2%, 5-year of two group were 59.4% and 58.3%, 10-year of two group were 50.8% and 55.3% respectively (P > 0.05). Meanwhile, the incidence of fever, infection, bleeding, secondary cancer between two groups was not significant different (P > 0.05). It is concluded that the hematopoietic reconstitution of high-dose chemotherapy/radiotherapy preconditioning group is later than that of high-dose chemotherapy preconditioning group. However, there is no significant difference in curative effect and prognosis between the two groups.
Adolescent ; Adult ; Aged ; Child ; Combined Modality Therapy ; Female ; Hematopoietic Stem Cell Transplantation ; Humans ; Lymphoma ; surgery ; therapy ; Male ; Middle Aged ; Prognosis ; Retrospective Studies ; Transplantation Conditioning ; methods ; Transplantation, Autologous ; Young Adult

This study was aimed to investigate the differences of therapeutic efficiencies, side effects and recovery rates of immune function in refractory lymphoma patients treated with autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT), autologous bone marrow transplantation (ABMT) and combination of APBHSCT with ABMT (APBHSCT + ABMT) by retrospective analysis, and to evaluate the merits and demerits of 3 kinds of transplantation for treatment of refractory lymphoma. 68 patients with malignant lymphoma were treated with autologous hematopoietic stem cells transplantation. Out of 68 patients 10 cases were treated with autologous bone marrow transplantation (ABMT), 46 cases were treated with autologous peripheral blood hematopoietic stem cell transplantation (APBHSCT), and 12 cases were treated with autologous peripheral blood hematopoietic stem cells transplantation combined with autologous bone marrow transplantation (APBHSCT + ABMT). The results indicated that the therapeutic response rates and survival rates at 1, 3, 5 years for each transplant regimen were 90% and 75%, 57.1%, 33.3%; 86.4% and 74.4%, 54.2%, 38.1%; 83.3% and 72.7%, 55.6%, 40%. The times of ANC > or = 0.5 x 10(9)/L were 13, 11 and 9 days, times of platelet >/= 20 x 10(9)/L were 17, 14 and 10 days. The recovery rates of T cell subtypes in patients received ABMT, APBHSCT and APBHSCT + ABMT on 3 months, 6 months, 1 year were (0%, 33.3%, 60%), (10.8%, 32%, 73.9%), (27.3%, 55.6%, 85.7%) respectively. In conclusion, the efficacy and side effects of APBHSCT + ABMT as compared with ABMT and APBHSCT are roughly the same, but ABMT + APBHSCT can result in more rapid hematopoietic reconstitution and less restrictions with contributes to widen choice of transplant regimen for patients with alder age and impaired hematopoietic functions.
Adolescent ; Adult ; Bone Marrow Transplantation ; Female ; Humans ; Lymphoma ; surgery ; Male ; Middle Aged ; Peripheral Blood Stem Cell Transplantation ; Retrospective Studies ; Transplantation, Autologous ; Young Adult

This paper explored the curative effect of combined modality therapy and extended field radiotherapy for early-stage Hodgkin's Lymphoma. 104 cases of early-stage Hodgkin's Lymphoma from Jan 1987 to Dec 2010 in PLA Hospital 307 were retrospectively analyzed, including 76 cases in combined modality therapy group and 28 cases in extended field radiotherapy group, and the long-term efficacy and toxicity of two therapy modalities were evaluated. The results showed that the median survival time of 104 cases was 85.42 months, the complete remission rates of combined modality therapy and extended field radiotherapy groups were 72.4 and 71.4 respectively (P = 0.924); the overall response rates of combined modality therapy and extended field radiotherapy groups were 97.4 and 96.4 respectively (P = 0.779); the 5-year overall survival (OS) rates in the 2 groups were 89.5 and 89.1 respectively, and the 8-year OS rates of the 2 groups were 81.3 and 70.6. No statistical difference was found in above-mentioned 2 groups. Moreover, the 5-year progression free survival (PFS) rates of these 2 groups were 84.2 and 69.0 (P = 0.04), and 8-year PFS rates of these 2 groups were 80.0 and 55.5 (P = 0.04) respectively, the 5-year relapse rates of these 2 groups were 28.1 and 45.6 (P = 0.023) respectively. It is concluded that the combined modality therapy can raise the PFS rate and reduce the relapse rate as compared with extended field radiotherapy for early-stage Hodgkin's Lymphoma, but there is no difference in the overall survival rate between the 2 groups.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antineoplastic Combined Chemotherapy Protocols ; Child ; Combined Modality Therapy ; Female ; Hodgkin Disease ; drug therapy ; radiotherapy ; Humans ; Male ; Middle Aged ; Retrospective Studies ; Survival Rate ; Treatment Outcome ; Young Adult

OBJECTIVETo study the effect of co-blockage of vascular endothelial growth factor (VEGF) and its receptor (KDR) on growth of bladder carcinoma T24 cells and nude mice xenograft.

METHODST24 cell line co-transfected with VEGF siRNA and sKDR expression plasmids was developed and its proliferation was assayed by MTT and apoptosis by FCM. The nude mice model bearing bladder carcinoma xenograft was established. The tumor cell VEGF expression, stroma microvessel density (MVD) and tumor cell topoisomerase II alpha (Topo II alpha) expression were detected by immunohistochemistry. Cell apoptosis was estimated by TUNEL assay.

RESULTSMTT assay showed that cell proliferation in VEGF siRNA, sKDR and combination groups was 56.3% +/- 8.3%, 42.6% +/- 13.8% and 32.5% +/- 4.3%, respectively, significantly lower than that in the scramble control (97.3% +/- 11.6%, P < 0.0001). FCM showed there were sub-diploid apoptotic peaks before G1 phase in VEGF siRNA, sKDR and combination groups, and apoptosis ratio was 5.1% +/- 0.9%, 4.2% +/- 0.5% and 8.8% +/- 0.7%, respectively, all of which were higher than that in the scramble control (0.9% +/- 0.4%, P < 0.05), and the combination group had even more higher apoptosis than the two singlely treated groups (P < 0.01). In vivo test showed that tumor growth was inhibited in VEGF siRNA, sKDR and combination groups, and from day 16 the tumor volume in combination group was significantly smaller than that in scramble control (P < 0.05), and from day 28 the tumor almost lost the ability to further growth. Immunohistochemistry revealed VEGF expression in combination group was 54.37 +/- 5.28, significantly lower than that in the scramble control (141.66 +/- 8.59, P < 0.0001). MVD number was only 8.22 +/- 3.79, much less compared with that in the scramble control (61.76 +/- 5.28, P < 0.0001) or sKDR group (19.46 +/- 4.16, P = 0.0089). Tumor cell proliferation index in the combination group (1.5% +/- 0.7%) was significantly decreased compared with that in the scramble control (11.8% +/- 5.2%, P < 0.0001), and apoptosis index (67.2% +/- 8.5%) was much higher than that in the scramble control (8.7% +/- 2.7%, P < 0.0001), VEGF siRNA group (54.3% +/- 4.8%, P = 0.0492) or sKDR group (52.3% +/- 6.4%, P = 0.0293).

CONCLUSIONVEGF siRNA or sKDR alone can inhibit tumor cell proliferation and induce cell apoptosis, but co-blockage of VEGF and KDR by their combination shows more significant therapeutic efficacy.

Animals ; Apoptosis ; Cell Line, Tumor ; Cell Proliferation ; Gene Expression Regulation, Neoplastic ; Humans ; Male ; Mice ; Mice, Inbred BALB C ; Mice, Nude ; Neoplasm Transplantation ; Neovascularization, Pathologic ; prevention & control ; Plasmids ; RNA Interference ; RNA, Small Interfering ; genetics ; Transfection ; Tumor Burden ; Urinary Bladder Neoplasms ; metabolism ; pathology ; Vascular Endothelial Growth Factor A ; genetics ; metabolism ; Vascular Endothelial Growth Factor Receptor-2 ; genetics ; metabolism ; Xenograft Model Antitumor Assays