Objective　To study the effects of recombinant BCG-Sj26GST vaccine of schistosoma japonicum of TNF-α and IL-1 produced by spleen cells of mice.Methods　In experiment 1,BALB/C mice were immnized subcutaneously by 106 and 108 CFU vaccine respectively,challenged with Sj cercariae on 8 w of immunization and killed on 6 w of infection to separate spleens,PBS served as control;in experiment 2,after immunized subcutaneously and intravenously by 106 CFU vaccine respectively,4 mice were killed to separate sleens on 0,4,8,10,14 and 16 w of immunization,spleen cells were stimulated by Sj26 or mitogens,the level of TNF-α in supernatant of spleen cells were detected by ELISA,that of IL-1 by fibroblast proliferative response.Results　Levels of TNF-α and IL-1 increased obviously by immunization with the vaccine against challenge with Sj cercariae;dynamic observation showed that TNF-α and IL-1 in the subcutaneous group reached the highest level on 4～8 w;that in the intravenous group on 16 w and 4～8 w respectively.Conclusions　Recombinant BCG-Sj26GST vaccine of schistosoma japonicum may promote spleen cell of mice to secrete TNF-α and IL-1 early,these cytokines may play an important role in protective immunity against schistosomiasis.

Adenocarcinoma ; genetics ; pathology ; surgery ; Adult ; Aged ; Aged, 80 and over ; Biopsy ; methods ; Bronchoscopy ; Carcinoma, Large Cell ; genetics ; pathology ; surgery ; Carcinoma, Non-Small-Cell Lung ; genetics ; pathology ; surgery ; Carcinoma, Squamous Cell ; genetics ; pathology ; surgery ; Female ; Gene Amplification ; Humans ; Lung Neoplasms ; genetics ; pathology ; surgery ; Male ; Middle Aged ; Polyploidy ; Receptor, Epidermal Growth Factor ; genetics ; Young Adult

Carcinoma, Non-Small-Cell Lung ; metabolism ; pathology ; Cell Membrane ; metabolism ; Cell Nucleus ; metabolism ; Cytoplasm ; metabolism ; Humans ; Lung Neoplasms ; metabolism ; pathology ; Signal Transduction ; TCF Transcription Factors ; metabolism ; Transcription Factor 7-Like 2 Protein ; Wnt Proteins ; physiology ; beta Catenin ; metabolism

OBJECTIVETo observe the intervention effects of electroacupuncture at "Yishu" (EX-B 3) on rats with type-2 diabetes mellitus (T2DM), so as to provide experiment references for acupuncture to treat T2DM.

METHODSAmong seventy male Wistar clean-grade rats, 8 rats were randomly selected into a control group; the rest rats were made T2DM model. Fifty-two rats which were successfully made T2DM model, according to randomized block method, were divided into a model group (10 rats), a medication group (10 rats), an electroacupuncture at "Shenshu" (BL 23) group (11 rats), an electroacupuncture at "Pishu" (BL 20) group (10 rats) and an electroacupuncture at "Yishu" (EX-B 3) group (11 rats). Seven days after successful establishment of model, the rats in the model group were fixed in the self-made rat bag without receiving any treatment; the rats in the medication group, according to body mass (10 mL/kg), were treated with intragastric administration of glimepiride; the rats in all the electroacupuncture groups were treated with electroacupuncture at "Shenshu" (BL 23), "Pishu" (BL 20) and "Yishu" (EX-B 3), respectively. The continuous wave was selected with a frequency of 15 Hz and a current intensity of 4 to 6 mA. The treatment was given 20 min per treatment, once a day, 5 treatments per week for continuous 4 weeks. Before the establishment of model and continuous 4 weeks after the intervention, blood samples were collected from rats' caudal vein, and fasting blood glucose (FBG) was measured with FBG device each week. After the last intervention, the rats were killed and hypothalamus, pituitary and adrenal gland were collected. The colorimetric method was applied to measure the contents of triglyceride (TG), cholesterol (TC), high-density lipoprotein (HDL-C) and low-density lipoprotein (LDL-C); radioimmunoassay was used to test the contents of glycated serum protein (GSP), fasting insulin (FINS), corticotropin-releasing hormone (CRH), adrenocorticotropic hormone (ACTH) and cortin (CORT).

RESULTSFour weeks after the intervention, except that the rat's body mass in the normal group continued to increase, body mass in the model group, medication group and each electroacupuncture group were significantly reduced (P<0.01, P<0.05). Compared with the model group, the FBG in the electroacupuncture at "Pishu" (BL 20) group and electroacupuncture at "Yishu" (EX-B 3) group were obviously reduced (P<0.05, P<0.01); FBG in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (both P<0.05). The contents of TG, HDL-C and LDL-C in the electroacupuncture at "Yishu" (EX-B 3) group were reduced (P<0.05, P<0.01), the content of TG was significantly lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (both P<0.05), the content of LDL-C was significantly lower than that in electroacupuncture at "Shenshu" (BL 23) group (P<0.05). Insulin sensitivity index (ISI) in the medication group, electroacupuncture at "Pishu" (BL 20) group and electroacupuncture at "Yishu (EX-B 3)" group were evidently increased (P<0.05, P<0.01); ISI in the medication group was lower than that in the electroacupuncture at "Yishu" (EX-B 3) group (P<0.05). The content of CRH in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Shenshu" (BL 23) group (P<0.05, P<0.01); the content of CORT in the electroacupuncture at "Yishu" (EX-B 3) group was lower than that in the medication group and electroacupuncture at "Pishu" (BL 20) group (both P<0.05).

CONCLUSIONElectroacupuncture at "Yishu" (EX-B 3) could reduce the level of CORT to improve the insulin resistance in rats with T2DM, improve insulin sensitivity index, regulate blood lipid metabolism and relieve the hyperactivity of the HPA axis.

Acupuncture Points ; Acupuncture Therapy ; Animals ; Blood Glucose ; metabolism ; Diabetes Mellitus, Type 2 ; metabolism ; therapy ; Electroacupuncture ; Hormones ; metabolism ; Humans ; Hypothalamus ; metabolism ; Male ; Pituitary-Adrenal System ; metabolism ; Rats ; Rats, Wistar

Artemisnin is a novel sesquiterpene lactone with an internal peroxide bridge structure, which is extracted from traditional Chinese herb Artemisia annua L. (Qinghao). Recommended by World Health Organization, artemisinin is the first-line drug in the treatment of encephalic and chloroquine-resistant malaria. In the present study, transgenic A. annua plants were developed by overexpressing the key enzymes involved in the biosynthetic pathway of artemisinin. Based on Agrobacterium-mediated transformation methods, transgenic plants of A. annua with overexpression of both HDR and ADS were obtained through hygromycin screening. The genomic PCR analysis confirmed six transgenic lines in which both HDR and ADS were integrated into genome. The gene expression analysis given by real-time quantitative PCR showed that all the transgenic lines had higher expression levels of HDR and ADS than the non-transgenic control (except ah3 in which the expression level of ADS showed no significant difference compared with control); and the HPLC analysis of artemisinin demonstrated that transgenic A. annua plants produced artemisinin at significantly higher level than non-transgenic plants. Especially, the highest content of artemisinin was found in transgenic line ah70, in which the artemisinin content was 3.48 times compared with that in non-transgenic lines. In summary, overexpression of HDR and ADS facilitated artemisinin biosynthesis and this method could be applied to develop transgenic plants of A. annua with higher yield of artemisinin.
Artemisia annua ; genetics ; metabolism ; Artemisinins ; metabolism ; Biosynthetic Pathways ; Drugs, Chinese Herbal ; Mixed Function Oxygenases ; genetics ; Oxidoreductases ; genetics ; Plant Proteins ; genetics ; Plants, Genetically Modified ; genetics ; metabolism ; Plants, Medicinal ; genetics ; metabolism

OBJECTIVETo study the effective method of microsurgical resection of the recurrent craniopharyngiomas.

METHODSMicrosurgical resection underwent in 40 cases with the recurrent craniopharyngiomas that accounted for 24 men and 16 women with mean age 35.1 year old. Visual deterioration was mainly complained in 25 patents, headache of symptoms in 9 patients, defect of visual field in 2 patients, amenorrhea in 2 patients, hyposexuality in one patient and diabetes insipidus in the one. Average history was 2.9 years. The superior sellar tumors on MR imaging grew in 19 cases, the superior-inferior sellar mass in 9 cases, intra-sellar in 5 cases and the tumors into the third ventricle in 7 cases. The huge calcification tumors were found in 5 cases, cystic tumors in 21 cases, and solid tumors 2 cases. Hydrocephalus presented in 12 cases. Evolution of tumor residuum was revealed in 31 cases after initial surgery and recurrent tumor after completing total removal of the tumor in 9 cases. The pterional approach was employed in 33 cases. The longitudinal fissure to the corpus callus approach in 2 cases was available for resection of the third ventricular tumor through the fornix column and septum pellucidum spaces.

RESULTSIn 33 cases with the pterional approaches, total removal of the tumors were completed in 22 cases, subtotal removal of tumors in 9 cases, and partial removal in 2 cases. In 5 cases with subfrontal approach, 4 patients the total removal of tumors obtained in 4 cases and one subtotal removal of tumor in one. Of 2 cases with the longitudinal fissure to the corpus callus approaches, one case was achieved with the total removal of tumor and the other with subtotal removal of tumor. The pituitary stalk was preserved in 8 cases, the pituitary stalk was severed in 9 cases and the pituitary stalk was not identified in 23 cases when surgery. 17 patients experienced diabetes insipidus and 12 patients had the hypothalamic hypofunction after surgery. One death occurred from the hypothalamic hypofunction 35 day after surgery. By follow-up from 3 months to 3 years, 22 patients returned normal life, 11 patients can carry out their self-life, and 6 patients need care.

CONCLUSIONSThe desirable removal of recurrent craniopharyngioma could be completed in the majority of patients although the reoperation of the tumors was performed very difficulty owing to the tumor adhesive to the surrounding hypothalamic structures.

Adolescent ; Adult ; Child ; Craniopharyngioma ; surgery ; Female ; Humans ; Male ; Microsurgery ; Middle Aged ; Neoplasm Recurrence, Local ; surgery ; Neurosurgical Procedures ; methods ; Pituitary Neoplasms ; surgery ; Reoperation ; Retrospective Studies ; Treatment Outcome

The present study is to investigate the quality changes of ginseng stems and leaves before and after frost. The contents changes of ginsenoside, free amino acid, and total phenolic compounds, as well as DPPH radical scavenging effect before and after frost were measured. The content of 9 ginsenoside monomer in ginseng stems was decreased except for Rg, and Re after frost, but in ginseng leaves was all decreased. The total content of amino acids was decreased in ginseng stems after frost, while increased in ginseng leaves. The content of phenolic compounds in ginseng stems and leaves were both decreased after frost while the ability of DPPH radical scavenging was improved. The factor of frost has great impact on the quality of ginseng stems and leaves.
Drugs, Chinese Herbal ; chemistry ; Ecosystem ; Freezing ; Panax ; chemistry ; Plant Leaves ; chemistry ; Plant Stems ; chemistry ; Quality Control

Objective To explore the role of NF-kB activation on spontaneous formation of germinal centers in spleen in BXSB mice and it's mechanisms.Methods Eighteen BXSB mice were divided to control group and pyrrolidine dithiocarbonate(PDTC)group randomly.PDTC group was given PDTC 120 mg/kg?BW ip every other day and control group was given the same dose of dissolving solution.NF-kB activity was deter- mined by electrophoretic mobility shift assay.Two color flow cytometry were used to detect CD154 expression on splenic B cells and germinal center B cells apoptosis.Germinal centers were stained for histochemical analysis.Results PDTC could inhibit the NF-kB activity in spleen tissue in BXSB mice.It decreased the NF-kB activity by 62.82%.Spontaneous germinal center formation was detected in spleen in BXSB mice.In- hibiting NF-KB activation could down-regulate CD154 expression on splenic B cell,retard spontaneous germi- nal center formation and increase germinal center B cell apoptosis.Conclusion NF-kB activation may induce spontaneous germinal center formation in spleen in BXSB mice by upregulating CD154 expression on splenic B cell and decreasing germinal center B cell apoptosis.The autoreactive B cells generated during spontaneous germinal center formation may escape apoptosis and then differentiate to autoantibody-producing plasm cells.It suggests that NF-kB can be a therapeutic target.

Genome duplication in E. coli is carried out by DNA polymerase III, an enzyme complex consisting of ten subunits. Investigations of the biochemical and structural properties of DNA polymerase III require the expression and purification of subunits including α, ge, θ, γ, δ', δ, and β separately followed by in vitro reconstitution of the pol III core and clamp loader. Here we propose a new method for expressing and purifying DNA polymerase III components by utilizing a protein co-expression strategy. Our results show that the subunits of the pol III core and those of the clamp loader can be coexpressed and purified based on inherent interactions between the subunits. The resulting pol III core, clamp loader and sliding clamp can be reconstituted effectively to perform DNA polymerization. Our strategy considerably simplifies the expression and purification of DNA polymerase III and provides a feasible and convenient method for exploring other multi-subunit systems.
Cloning, Molecular ; DNA Polymerase III ; chemistry ; genetics ; metabolism ; DNA Replication ; DNA, Bacterial ; biosynthesis ; genetics ; Escherichia coli ; enzymology ; genetics ; Plasmids ; metabolism ; Polymerization ; Protein Engineering ; methods ; Protein Subunits ; chemistry ; genetics ; metabolism ; Recombinant Proteins ; chemistry ; genetics ; metabolism

OBJECTIVEThe etiology of developmental dislocation of the hip (DDH) remains uncertain, but some research has shown that this disorder is closely related to hip joint laxity. This study examined the expression of collagens type I and III mRNA and protein in the hip capsule of children with DDH in order to investigate the roles of collagens type I and III in hip joint laxity.

METHODSNine children with DDH and nine age and gender-matched normal children (control group) were enrolled. Semiquantitative RT-PCR method was used to detect mRNA expression of COL1a1 and COL3a1 in the hip capsule. Western-Blot method was used to detect protein expression of COL1a1 and COL3a1 in the hip capsule. The quantitative analysis of the COL1a1 and COL3a1 was performed by professional image software and the results were analyzed with standard statistical methods.

RESULTSmRNA and protein expression of COL1a1 in the DDH group was significantly lower than that in the control group (P<0.01). Compared with the control group, COL1a3 mRNA expression in the DDH group decreased significantly (P<0.01), but COL1a3 protein expression was not significantly different.

CONCLUSIONSThe decreased collagen I mRNA and protein expression in the hip capsule might contribute to hip joint laxity in children with DDH. Collagen type III may not be associated with hip joint laxity in DDH.

Blotting, Western ; Child ; Child Development ; Child, Preschool ; Collagen Type I ; analysis ; genetics ; Collagen Type III ; analysis ; genetics ; Female ; Hip Dislocation ; metabolism ; Humans ; Infant ; Male ; RNA, Messenger ; analysis ; Reverse Transcriptase Polymerase Chain Reaction