To determine the number average molecular mass of chitosan-oligosaccharide by u-sing the acetylacetone's method based on the color-producing reaction with amino terminal . Via the tests on the repetitiveness, accuracy and the stability, supposed this method was reliable. At the same time, the contrastive results of HPLC and the traditional K3Fe(CN)6 method indicate that the acetylacetone method was more suitable for the determination of chitosan-oligosaccharide molecular mass.

Angiogenesis involves the occurrence and development of serious diseases such as cardiovascular and cerebrovascular diseases and malignant tumors. Promoting or inhibiting angiogenesis is considered to be a key factor in controlling the progression of the disease. Chinese materia medica (CMM) has become a research hotspot because of its good clinical therapeutic effect and significant intervention on angiogenesis. The zebrafish model as an ideal animal model also provides a good experimental basis for the study of angiogenesis. CMM monomers, extracts, prescriptions and Chinese patent medicine, and other forms of intervention were selected as the research object. Based on zebrafish animal model experiments, we summarize domestic and foreign relevant research and review the research progress of the effect of CMM on angiogenesis in zebrafish, aiming to provide reference for further development and application of zebrafish as a model and further study of CMM intervention in angiogenesis.

The present condition in the acupoint term translation was analyzed and its existent problems in this area were discussed in this paper. The authors suggested that in translating the terms of acupoints, the translation on the meaning of the acupoints should be added, in this way, it can not only keep the integrity in acupoint translation, but also make the inheritance of the Chinese precious culture of Traditional Chinese Medicine further available.
Acupuncture Points ; Animals ; Culture ; Humans ; Medicine, Chinese Traditional ; Translating

OBJECTIVE Metastasis-associated in colon cancer-1 (MACC1) is an oncogene that has been newly identified. It promotes tumor proliferation and invasion via the MET pathway. Our study investigated the effects of Saikosaponin-b(SS-b) on the proliferation and apoptosis of HepG2 cells and its regulation on MACC1/c-Met/Akt signaling pathway. METHODS HepG2 cells were treated with SS-b (10-800 g·L-1) for 48 h in vitro. The CCK-8 assay was used to assess cell proliferation, and cell apoptosis was determined by Hoechst33258 staining, AnnexinⅤ/PI staining and caspase 3 assay. RT-PCR was used to examine the expression of MACC1, c- MET and hepatocyte growth factor (HGF) mRNA. MACC1 protein was detected by Western blot and immunohistochemistry. The protein expressions of p-c-MET, c-MET, p-AKT, AKT, p-BAD, BAD were measured by Western blot. RESULTS SS-b inhibited the growth of HepG2 cells in dose-dependent way and induced cell apoptosis significantly. HepG2 cells showed karyopyknosis, fragmentation and fluorescence highlight in SS-b treatment group. FCM results showed that apoptosis rate of HepG2 cells increased with SS- b concentration. The immunofluores?cence results showed that the MACC1 expression decreased significantly in HepG2 cells treated with SS-b. The expression levels of MACC1, c-MET and HGF mRNA in HepG2 cells were significantly inhibited by SS-b. SS-b also significantly decreased the protein expressions of MACC1, p-c-MET and p-AKT while increased the expression of p-BAD and caspase 3 in HepG2 cells(P<0.05). CONCLUSION SS-b inhibited the proliferation and induced the apoptosis of HepG2 cells by targeting the MACC1/c-Met/Akt signaling pathway.

OBJECTIVE Angiogenesis therapy has attracted interest as a potential treatment for hepatocellular carcinoma (HCC). In this study, we investigated the anti-proliferative activities and anti-angiogenesis effects of saikosaponins (SS)-b on hepatocellular carcinoma (HCC) and its regulation on VEGF/ERK/HIF-1α signal pathway. METHODS H22 hepatoma-bearing mice model and HepG-2 cells were used to study the anti-tumor and anti-angiogenesis effects of SS-b in vivo and in vitro. Pathological change of tumor tissue was observed by HE staining, the microvascular changes were detected by immunohistochemical method. The effects of SS-b on angiogenesis were examined by using the chick embryo chorioallantoic membrane (CAM) model. The effects of SS- b on proliferation, migration and invasion were investigated by MTT assay, scratch wound healing assay and transwell assay inhuman umbilical vein endothelial cell (HUVEC) and HepG2 cells in vitro. Vascular endothelial growth factor (VEGF), matrix metalloproteinase-2/9(MMP-2/9), hypoxia-inducible factor-1α (HIF-1α) expression and the phosphorylation of extracellular regulated kinase(ERK) were analyzed using RT-PCR and Western-blot. RESULTS SS-b effectively inhibited the tumor growth of H22 mice in vivo. The inhibitory rate of tumor was 49.1%, 50.7%, 66.1% in SS-b 5, 10 and 20 mg·kg-1 group respectively. HE staining results showed that SS-b induced tumor necrosis and nuclear dissolution in H22 mice. Moreover, SS-b also reduced the number of microvessels of tumor tissue in H22 mice significantly and suppressed the angiogenesis of CAM induced by b-FGF. SS-b had an obvious inhibitory effect on cell proliferation, migration and invasion of HUVEC cells and HepG-2 cells. These effects were associated with down-regulation of the expression of MMP2/9 and suppression of VEGF/ERK/HIF-1α signaling in H22 mice and Hep-G2 cells. CONCLUSION Our findings showed that SS-b exerts anti-tumor effects by inhibit?ing tumor angiogenesis via regulating VEGF/ERK/HIF-1α signal pathway in vivo and in vitro.

LRP16 is a novel gene which was found in our laboratory by using methylation-sensitive restriction landmark genomic scanning (RLGS) technique. In order to clone the full-length cDNA of this leukemia relapse associated gene, the method of rapid amplification of cDNA end (RACE) was employed. By optimizing some procedures of RACE method, the 5'- and 3'-untranslated region of LRP16 cDNA was successfully sequenced. Then, the full length of LRP16 cDNA and open reading frame (ORF) was constructed and was registered in GenBank. The above-mentioned procedure demonstrated RACE technique is a rapid and sensitive method for cloning unknown gene. Especially, it is very useful to cloning the 5'- and 3'-untranslated region of a novel gene.

A fermentation system composed of four airlift suspended-bed bioreactors in series and with a total working volume of 4800 mL was established. Continuous ethanol fermentation using self-flocculating yeast SPSC01, a fusant from Saccharomyces cerevisiae and Schizosaccharomyces pombe, and two-stage enzymatic hydrolyte of dry milling corn powder, was continuously run for 120 days. All of the backset distillage collected after distilling the final beer was used to mix the corn powder and no any other wastes except the solid residue of corn powder was discharged from the fermentation system, which guaranteed the distillage to be recycled at its maximum. The experimental results revealed that both ethanol and residual sugar in the final beer could be maintained relatively stable with their average levels of 93.6 and 7.9 g/L, respectively when the fermentation system was operated at the dilution rate of 0.05 h(-1). Parameter oscillations reported previously were also observed for the first and second bioreactors, but were effectively attenuated thereafter, which indicated that high yeast cell concentrations resulted from the self-immobilization of this special self-flocculating strain contributed to damp these oscillations. The monitoring of residual nitrogen and phosphor indicated that the accumulations of these nutritional elements occurred and the amount of these inorganic salts supplemented in the substrate should be decreased properly.
Bioreactors ; microbiology ; Ethanol ; metabolism ; Fermentation ; Flocculation ; Immobilization ; Saccharomyces cerevisiae ; metabolism ; Schizosaccharomyces ; metabolism ; Zea mays ; metabolism

Objective To construct small RNA deletion and overexpression strains with a length of less than 100 nt in Yersinia pestis.Methods Deletion mutants of the target sRNAs were constructed by increasing the length of homologous regions.Meanwhile, the high copy plasmid pBAD/HisA was modified into an inducible transcriptional vector as an sRNA-overexpression plasmid by using QuikChange lightning site-directed mutagenesis kit .The presence , size, and transcription-al initiation sites of the indicated sRNA were predicted by transcriptome sequencing , primer extension , and previous stud-ies.The full-length DNA fragments of target sRNAs were transformed into the transcriptional vector .The overexpressing strains of sRNAs were identified by Northern Blot .Results and Conclusion Four sRNAs deletion mutants of sR01, sR02, sR03 and HmsA and three sRNAs overexpression mutants MicF , HmsA and CpxQ were successfully constructed .A method of construction of sRNA deficient and overexpressing strains of Y.pestis has been quickly and efficiently established by λ-Red homologous recombination technology and QuikChange ? lightning site-directed mutagenesis kit.

OBJECTIVESTo investigate the oncologic outcomes of surveillance, retroperitoneal lymph node dissection (RPLND) and primary chemotherapy in patients with clinical stage Ia nonseminomatous germ cell testicular tumors (CS Ia NSGCT) and to analyze risk factors for relapse.

METHODSPatients with CS Ia NSGCT were retrospectively reviewed. Totally 72 patients were enrolled and grouped according to three different treatment after orchiectomy, among them 33 cases in surveillance group, 24 cases in RPLND group and 15 cases in primary chemotherapy group. Disease progressive free survival and disease specific survival were compared using Kaplan-Meier analysis. Cox regression analysis was used to confirm variables those were associated with disease progression.

RESULTSAll 72 patients were followed-up at mean 62 months (12 - 175 months), 6 patients had evidence of relapse. Both the 5-year disease specific survival and 5-year overall survival rate were 100%. For surveillance, chemotherapy and RPLND, cumulative 5-year PFS rates were 84.0%, 93.3% and 100%, respectively. Relapse rate was higher in surveillance group than in RPLND group (17.8% vs. 0, χ² = 3.99, P = 0.04). Patients with the history of cryptorchidism also have higher relapse rate than without (37.5% vs. 4.7%, χ² = 10.02, P = 0.01). In the surveillance cohort, relapse rates were significantly higher in patients with a predominant component of embryonal carcinoma (3/6 vs. 7.4%, χ² = 6.93, P = 0.04) and for those over 13 years of age (23.1% vs. 5.3%, χ² = 4.33, P = 0.04). On multivariate analysis, treatment mode of patients (OR = 0.08, 95% CI: 0.06-0.36, P = 0.03) and patients with a history of cryptorchidism (OR = 25.3, 95% CI: 6.57-78.42, P = 0.04) were independent predictors of relapse.

CONCLUSIONSSurveillance, RPLND and adjuvant chemotherapy could be reliable strategies in compliant stage Ia nonseminoma patients and achieve satisfactory overall survival. Relapse rate is relatively higher for patients with surveillance. Those who are older or have a history of cryptorchidism experience a higher risk of relapse.

Adolescent ; Adult ; Aged ; Chemotherapy, Adjuvant ; Child ; Child, Preschool ; Humans ; Infant ; Kaplan-Meier Estimate ; Male ; Middle Aged ; Neoplasm Recurrence, Local ; Neoplasms, Germ Cell and Embryonal ; therapy ; Orchiectomy ; Postoperative Period ; Retrospective Studies ; Risk Factors ; Survival Rate ; Testicular Neoplasms ; therapy ; Treatment Outcome ; Young Adult

Aged ; Antineoplastic Agents ; administration & dosage ; Cystectomy ; Humans ; Infusions, Intra-Arterial ; Male ; Neoplasm Recurrence, Local ; Pelvic Neoplasms ; drug therapy ; Urinary Bladder Neoplasms ; surgery