[Effects of moxibustion at "Tianshu"（ST25） and "Shangjuxu" （ST37） on colonic metabolites and inflammatory factors in rats with Crohn's disease].

OBJECTIVE: To observe the effects of moxibustion at "Tianshu"（ST25） and "Shangjuxu"（ST37） on the colonic metabolites and inflammatory factors in rats with Crohn's disease（CD）, so as to explore the mechanisms of moxibustion in protecting colon of CD rats based on metabolomics. METHODS: Twelve rats were first randomly selected from 36 male SD rats as a normal group（NG）. The CD model was induced by 2, 4, 6 trinitrobenzene sulfonic acid（TNBS） enema on the rest 24 rats. After successful modeling, rats were randomly divided into model（TNBS） and moxibustion（TNBS+MOX） groups（n=10 rats/group）. Moxibustion was applied at bilateral ST25 and ST37 for 30 min, once daily for 7 consecutive days in the TNBS+MOX group, while rats in the NG and TNBS groups did not receive any interventions. Body weight of rats was recorded and disease activity index（DAI） was assessed during the experiment. After interventions, HE staining was performed to observe pathological damage of colon. Serum levels of inflammatory factors were measured by ELISA. NMR hydrogen spectroscopy was used to detect colonic metabolites of each group, and orthogonal partial least squares discriminant analysis（OPLS-DA） was used to screen differential colonic metabolites between groups, followed by pathway analysis using MetaboAnalyst 5.0 platform. RESULTS: After modeling, compared with the NG group, the body weight of the rats in the TNBS group was significantly decreased（P<0.05）, the DAI score was increased （P<0.05）, the colon had obvious inflammatory damage and the pathological injury index was increased（P<0.05）, and levels of serum tumor necrosis factor-α（TNF-α）, interleukin（IL）-1ß and interferon-γ（IFN-γ） were significantly increased（P<0.05）. After moxibustion intervention, compared with the TNBS group, the body weight was significantly increased（P<0.05）, while the levels of serum TNF-α, IL-1ß, IFN-γ, and DAI score of the rats in the TNBS+MOX group were significantly decreased（P<0.05）, with alleviated colonic inflammatory injury detected by HE staining. Compared with the NG group, the relative expressions of colonic hypoxanthine, betaine, creatine, inositol, taurine, uracil, and methanol of the TNBS group were decreased（P<0.05）, while the relative expressions of histidine, leucine, proline, lysine, isoleucine, phenylalanine, tyrosine, propionic acid, and valine were increased（P<0.05） in the TNBS group, among which, relative expressions of hypoxanthine, leucine, lysine, isoleucine, betaine, tyrosine, and taurine were reversed in the TNBS+MOX group relevant to the TNBS group, mainly involving phenylalanine, tyrosine and tryptophan biosynthesis, and taurine and subtaurine metabolism pathway. CONCLUSION: The mechanism of moxibustion at ST25 and ST37 for CD may be related to improving colon metabolic disorder state by regulating multiple metabolic metabolites and metabolic pathways, and reducing the level of inflammatory factors, so as to maintain intestinal immune homeostasis.

Recombinant Human Granulocyte Colony-Stimulating Factor Promotes Preinvasive and Invasive Estrogen Receptor-Positive Tumor Development in MMTV-erbB2 Mice / 한국유방암학회지

PURPOSE: We investigated whether recombinant human granulocyte colony-stimulating factor (rhG-CSF) could promote the development of preinvasive and invasive breast cancer in mouse mammary tumor virus (MMTV-erbB2) mice with estrogen receptor-positive tumors. METHODS: MMTV-erbB2 mice were randomly divided into three experimental groups with 20 mice in each group. MMTV-erbB2 mice were treated with daily subcutaneous injections of vehicle or rhG-CSF (low-rhG-CSF group, rhG-CSF 0.125 microg; vehicle-rhG-CSF group, normal saline 0.25 microg; and high-rhG-CSF group, rhG-CSF 0.25 microg) at 3 months of age. Cellular and molecular mechanisms of G-CSF action in mammary glands were investigated via immunohistochemistry and reverse transcription polymerase chain reaction. RESULTS: Low, but not high, rhG-CSF doses significantly accelerated mammary tumorigenesis in MMTV-erbB2 mice. Short-term treatment with rhG-CSF could significantly promote the development of preinvasive mammary lesions. The cancer prevention effect was associated with reduced expression of proliferating cell nuclear antigen, cluster of differentiation 34, and signal transducers and activators of transcription 3 in mammary glands by >80%. CONCLUSION: We found that G-CSF was regulated by rhG-CSF both in vitro and in vivo. Identification of G-CSF genes helped us further understand the mechanism by which G-CSF promotes cancer. Low doses of rhG-CSF could significantly increase tumor latency and increase tumor multiplicity and burden. Moreover, rhG-CSF effectively promotes development of both malignant and premalignant mammary lesions in MMTV-erbB2 mice.

Comparative study on two types of 3S and P-loops digestive reconstruction after total gastrectomy for gastric cancer / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To compare the clinical effect of 3S-type and P-loops digestive reconstruction after total gastrectomy for gastric cancer.</p><p><b>METHODS</b>From February 2005 to February 2009, 85 cases underwent total gastrectomy in The First Affiliated Hospital of Henan University of Science and Technology. Two types of digestive reconstruction were performed with 3S-type jejunum(n=46) and P-loops Roux-en-Y esophagojejunostomy(n=39). The postoperative complications, nutrition index and the quality of life at half a year after surgery were comparatively analyzed.</p><p><b>RESULTS</b>Two types of digestive reconstruction had no statistical differences in operative time, postoperative complications and mortality(P>0.05). Compared with P-loops Roux-en-Y esophagojejunostomy at 6 months after operation, 3S-type jejunum had a lower incidence in dumping syndrome[4.3% (2/46) vs. 10.3% (4/39), P<0.05] and reflux esophagitis [10.8% (5/46) vs. 33.3% (13/39), P<0.05]. 3S-type jejunum was superior to P-loops Roux-en-Y esophagojejunostomy in serum total protein(55.7±3.1 g/L vs 50.3±5.1 g/L, P<0.05), albumin(36.5±3.6 g/L vs. 31.6±4.4 g/L, P<0.05), hemoglobin(120.2±13.4 g/L vs. 110.4±23.0 g/L, P<0.05), and nutritional assessment index(73.2±4.8 vs. 56.0±6.3, P<0.05).</p><p><b>CONCLUSION</b>Reconstruction of stomach with 3S-type jejunum may be an effective way to prevent reflux esophagitis and dumping syndrome, and to improve the nutritional status and the quality of life.</p>

Dendritic cells loaded with U251 tumor cell antigen enhance the cytotoxic effect of cytokine-induced killer cells against U251 cell line / 中华神经医学杂志

Objective To study the cytotoxic effect of cytokine-induced killer cells (CIKs) cocultured with U251 tumor cell antigen-loaded mature dendritic cells (DCs) against U251 cell line. Methods The DCs and CIKs were derived from the cord blood mononuclear cells (CBMCs) of the same donor. The DCs were challenged with U251 tumor cell antigen, and cocultured with the CIKs to induce the cell complex Ag-DC-CIK. The mature DCs were identified by morphological and phenotypic analyses. MTT assay was performed to detect the cytotoxic effects ofCBMCs, CIKs, antigen-loaded CIKs (Ag-CIK) or the cell complex Ag-DC-CIK in U25 ! cells. Results The mature DCs derived from the CBMCs highly expressed the costimulatory molecules CD86 (82.66%) and CD40 (69.40%), and moderately expressed CD83 (57.49%) and CD80 (51.14%). The cytotoxic activity of the cell complex Ag-DC-CIK against U251 cells (58.8%) was significantly higher than those of CBMCs (29.71%), CIKs (39.89%), and Ag-CIK (49.92%). Statistical analysis indicated significant difference in the cytotoxic activity between any two of the groups (P<0.05). Conclusion The DCs loaded with the tumor cell antigen can enhance the cytotoxic effect of the CIKs against the target tumor cells, which sheds light on a new approach of immunotherapy for intracranial tumors.

Expression and clinical significance of KiSS-1 and E-cadherin in gastric cardia carcinoma / 中华胃肠外科杂志

<p><b>OBJECTIVE</b>To investigate the expression and clinical significance of KiSS- 1 and E- cadherin in gastric cardia carcinoma and the correlation between the two proteins.</p><p><b>METHODS</b>The expression of KiSS- 1 and E- cadherin in 80 patients with gastric cardia carcinoma and 20 patients with normal gastric cardia epithelium was detected by immunohistochemical technique.</p><p><b>RESULTS</b>The expression of KiSS- 1 was negatively correlated with lymphatic metastasis and clinical stage (P < 0.05), but not correlated with the cancer differentiation (P < 0.05). The expression of E- cadherin was negatively correlated with lymphatic metastasis, clinical stage, and cancer differentiation (P < 0.05). Spearman test showed a positive correlation between KiSS- 1 and E- cadherin expression (r(s)=0.722, P < 0.05).</p><p><b>CONCLUSION</b>KiSS- 1 and E- cadherin may play important roles in inhibiting the invasion and metastasis of gastric cardia carcinoma.</p>

Isolation of 2,4-dichlorophenol degrading bacterium strain and cloning and expression of its 2,4-dichlorophenol hydroxylase gene / 生物工程学报

2,4-Dichlorophenol is toxic and biorefratory organic pollutant. A 2,4-dichlorophenol degrading bacterial strain GT241-1, identified as Pseudomonas sp., was isolated from soil samples which was collected from drainage area of several 2,4-dichlorophenol producing factories. Strain GT241-1 had strong 2,4-dichlorophenol degrading ability, it could decompose 91% 2, 4-dichlorophenol of 90 mg/L within 48 hours at 25 - 30 degrees C, and could utilize 2,4-dichlorophenol, 2,4-dichlorophenoxyacetic acid (2,4-D), benzoate and catechol as sole carbon and energy source. Southern blot showed that 2,4-dichlorophenol hydroxylase gene (dcpA) of strain GT241-1 locates on the about 10kb EcoR I/Xba I fragment. This fragment was recovered, linked to the vecter pUC19 and transformed into the E. coli DH5alpha. A aim transformant, Z539, was obtained by dot blotting from about 1200 transformants. PCR and the sequencing results shew that the whole dcpA gene is contained within the 10kb EcoR I /Xba I fragment of pZ539. This fragment was shortened to about 2.4kb by HindmIII. The shorted fragment was subcloned to vecter pRSET-B to get a transformant BS1-12. The subcloned fragment was sequenced. Sequencing results showed that the whole length of the subcloned fragment containing dcpA is 2389bp and the nucleotide span of coding region is from number 276 to number 2072 (1797 bp), with ATG and TAA as start and stop codon respectively. The sequence analysis of dcpA and the deduced amino acid encoded by dcpA showed that they are different from the relative sequences registered in the GenBank. The subcloned fragment carry the promoter of dcpA, this can deduce from the fact that the upflow length of dcpA coding region is 275bp, and further confirmed by the 2,4-dichlorophenol hydroxylase activity measurement results. The 2,4-dichlorophenol hydroxylase activity of transformant Z539 and BS1-12 were detected, the results showed these transformants have 2,4-dichlorophenol hydroxylase activity. By comparison, the activity of these transformants were lower than that of the strain GT241-1.

Gloning and Sequence Analysis of Dienelactone Hydrolase Gene / 微生物学通报

A 2,4 -dichlorophenol degrading Pseudomonas strain GI241-1 was isolated from a soil sample. The dienelactone hydrolase gene, designated as dcpD which encodes dienelactone hydrolase involved in transforming cis-2-chloro-dienelactone into 2-chloromaleylacetic acid, was cloned from this bacterium strain. The gene cloning strategy was to construct genomic library after location of its neighbouring gene by Southem blot and to screen the aim transformant by dot blotting. Sequencing results showed that length of dcpD is 702bp. The sequence of dcpD and the deduced amino acid are different from the relative sequences registered in the GenBank.