italic>γ-Aminobutyric acid (GABA) is a crucial inhibitory neurotransmitter found in various cells in the human body. While the GABAergic system is typically associated with the nervous system, recent research has revealed that immune cells and tumor cells also express components of this system. In the tumor microenvironment (TME), GABA is secreted to act extracellularly on other cells. GABA is metabolized via the GABA shunt and is involved in the tricarboxylic acid (TCA) cycle by generating succinate, which can provide energy for tumor cells. Activation of GABA receptors (GABARs) is a major pathway through which GABA participates in the regulation of antitumor immune responses. The activation of GABA type A receptors (GABA_ARs) can inhibit the activation and proliferation of T cells, elicit anti-inflammatory macrophages, and promote tumor cell growth and migration, while activation of GABA type B receptors (GABA_BRs) is generally considered to inhibit cancer cell migration and induce cancer cell apoptosis. In general, receptor activation inhibits immune cells, but the effect on tumor cells varies. Additionally, the downregulation of the expression levels of GABA transporters (GATs) is involved in tumor progression. Although antagonists of GABA metabolism and drugs that act on GABA receptors are considered therapeutic drugs for tumors, there have been few clinical studies conducted on them.

OBJECTIVETo develop a RP-HPLC method for the determination of the content of macrozamin in Rhizoma Heterosmilacis Japonicae.

METHODA Century C18 AQ column (4.6 mm x 250 mm, 5 microm) was used with the mobile phase consisted of water (4:96). The flow rate was 1.0 mL x min(-1). The detection wavelength was set at 215 nm, and the column temperature was 35 degrees C.

RESULTThe calibration curve was linear (r = 0.999 8) in the range of 19.12 - 382.4 microg x mL(-1) for macrozamin, the average recovery of the method was 99.5%, with RSD 2.1% (n = 9).

CONCLUSIONThis method can be used for the quality study of Rhizoma Heterosmilacis Japonicae.

Chromatography, High Pressure Liquid ; methods ; Liliaceae ; chemistry ; Methylazoxymethanol Acetate ; analogs & derivatives ; analysis ; Plants, Medicinal ; chemistry ; Reproducibility of Results ; Rhizome ; chemistry

Objective To determine the accuracy of femoral components sizing predicted by standardized digital templating in total knee arthroplasty (TKA).Methods Fifty consecutive patients (50 knees),who underwent primary TKAs for endstage osteoarthritis,were prospectively studied.The intra-operative and radiographic data were collected.All operations were performed by the same surgical techniques with PS type,open box Vanguard Complete Knee System.All patients underwent lateral and AP radiography of the involved knee under fluoroscopy before and after surgery.The distal femoral anteroposterior dimension (DFAP) were measured and the femoral components size were predicted on preoperative radiographs by two different methods:measurement of DFAP did not include (group A) the cartilage thickness of the medial posterior condyle or included that (group B).Cutting errors were corrected gradually,and DFAP was measured consequently.The most appropriate size was chose after each step respectively based on postoperative radiographs.The accuracy of femoral size predicted under different conditions was compared within two groups.Results During correction of cutting errors,the correct rate ranged from 18％ to 44％ in group A and from 26％ to 34％ in group B,the accuracy within one size ranged from 54％ to 84％ in group A and from 58％ to 84％ in group B.The cartilage thickness of medial posterior condylar,external rotation of femoral component,under-resected of anterior condylar,flexion of femoral component,and over-resected of posterior condylar can change the DFAP by 1.97±0.85 mm,1.56±2.06 mm,1.15±1.31 mm,-2.86±1.52 mm,and-0.87±0.77 mm,respectively.Conclusion Variation of intraoperative cutting errors and the cartilage thickness of medial posterior condyles can influence the accuracy of templating to some extent.Standardized digital radiography templating cannot predict femoral sizes accurately.

[Objective] To investigate the expression of MICA mRNA and protein in osteosarcoma and give a more comprehensive understanding to its immune evasion.[Methods] RT-PCR was used to analyze MICA mRNA in 11 osteosarcoma tissues and 5 osteosarcoma cell lines.MICA expression was examined in 66 paraffin-embedded osteosarcoma tissues and 6 paraffin-embedded normal bone tissues by immunohistochemistry,and MICA protein in 9 fresh osteosarcoma tissues was detected by Western blot too.MICA surface expression was estimated by flow cytometry.[Results] Nine of eleven (81.8%) osteosarcoma specimens and all of the five cell lines consistently expressed MICA mRNA.Up-regulation of MICA expression was found in osteosarcoma (34/66,51.6%),compared with normal bone tissues (0/6,0%).The cell lines Saos-2,MG63,and HOS showed positive surface MICA expression,while the U2OS and OS732 showed very limited expression.[Conclusion] MICA mRNA and protein predominantly expressed on osteosarcoma tissues and cell lines.

Objective To investigate the influence of Ethyl pyruvate(EP) on the level of high mobility group box 1 (HMGB1) in brain injury of infant rats induced by lipopolysaccharide (LPS) and its significance.Methods Two hundred and forty normal healthy 1-month-old Spragne-Dawley (SD) rats were randomly divided into 3 groups:9 g/L sodium chloride (NS) group(n=80),LPS group (n=80),and EP group (n=80).LPS (1 mg/kg) was injected via internal carotid.In EP group,after injecting LPS,each rat was immediately administrated 4 mL EP(40 mg/kg) intraperitoneally; in control group,4 mL Ringer's solution was given instead of EP.Rats were decapitated at 6,12,24,48 and 72 h following drug injection.The evan's blue (EB) content of brain tissues was meteraged by the formamide methods.Immunohistochemistry technology was used to detect the expression of HMGB1,neuron specific enolase(NSE) and glial fibrillary acidic protein(GFAP) protein,and reverse transcribe polymerase chain reaction technology was applied to study the expression level of HMGB1 mRNA in brain tissue.Meanwhile,the pathological changes of brain tissues were observed under the light microscope.Results Six hours after LPS was given,brain EB content,the levels of NSE,GFAP protein started to rise,reaching the peak at 24 h,and still higher than those in control group at 48 h and 72 h.The expression pattern of HMGB1 protein and mRNA in brain tissue was consistent with the severity of brain injury,increased at 6 h after LPS was given and reached the peak at 24 h,still higher than those in control group at 48 h and 72 h.Positive correlation was found among HMGB1 protein,HMGB1 mRNA and EB content,NSE protein,GFAP protein,respectively.In EP group,the levels of HMGB1 protein and mRNA,the levels of brain EB content,NSE,GFAP protein were found,positive correlation was still gotten between HMGB1 protein,EB content and NSE,GFAP protein,HMGB1 mRNA in LPS group and EP group.Conclusion EP has neuroprotective effect on brain injury induced by LPS,which may be relevant to decreasing of the expression of HMGB1 and suppressing inflammation action.

Objective To investigate the clinicopathologic relationship between serum platelet derived growth factor-BB(PDGF-BB)and IgA nephropathy in children.Methods The level of serum PDGF-BB was detected by double antibody enzyme linked immunosorbent assay(ELISA) in 15 cases healthy children and 30 cases IgA nephropathy.According to patholgical degree constituted by WHO in 1982,the IgA nephropathy group divided into 5 degrees:Ⅰ,Ⅱ,Ⅲ,Ⅳ,Ⅴ(Ⅰ,Ⅱ were light patholgic change group;Ⅲ,Ⅳ were moderate patholgic change group;Ⅴ was severe patholgic change group).The serum PDGF-BB in IgA nephropathy group and none-IgA nephropathy group,and in different renal pathology type IgA nephropathy group were analyzed.Data were analyzed by using SAS 6.12 software.Results The level of serum PDGF-BB were(247.35?55.79) ng/L in control group and(869.16?200.73) ng/L in IgA nephropathy group.It was higher in IgA nephropathy group than in control group,the difference was significant(P

The coding sequence for the mature peptide of porcine lactoferrin (Plf) was synthesized according to the codon usage of lactobacillus, to establish optimized porcine lactoferrin Lactobacillus expression system. The gene was ligated into the Xho I/BamH I site of Lactobacillus expression vector pPG612.1 and the recombinant plasmid pPG612.1-plf was transformed individually into Lactobacillus casei ATCC393, Lactobacillus pentosus KLDS1.0413, Lactobacillus plantarum KLDS1.0344 or Lactobacillus paracasei KLDS1.0652 by electroporation. After induction with xylose, expression of the recombinant proteins was detected by Western blotting and confocal laser scanning microscopy. Secretion of recombinant Plf proteins from four recombinant species was determined quantitatively by ELISA. The antibacterial activities of recombinant proteins were measured by agar diffusion method. The result shows that Plf was correctly expressed in four species of recombinant lactobacillus, with molecular weight of about 73 kDa. The expression levels in recombinant Lactobacillus casei, Lactobacillus pentosus, Lactobacillus plantarum, Lactobacillus paracasei were 9.6 μg/mL, 10.8 μg/mL, 12.5 μg/mL and 9.9 μg/mL, respectively. Antimicrobial activity experiment shows that the recombinant proteins were active against E. coli, Staphylococcus aureus, Salmonella typhimurium, Listeria, Pasteurella. The recombinant Plf expressed by recombinant Lactobacillus plantarum showed the best antibacterial activity among all recombinant lactobacillus species. These data represent a basis for the development and application of porcine lactoferrin from recombinant lactobacillus.
Animals ; Anti-Bacterial Agents ; biosynthesis ; Lactobacillus ; metabolism ; Lactoferrin ; biosynthesis ; Recombinant Proteins ; biosynthesis ; Swine

Objective To investigate the levels of miR-1 9a and miR-1 9b expressions in cancer tissues and serum of patients with non-small cell lung cancer (NSCLC),and evaluate the potential of miR-1 9a and miR-1 9b as biomarkers of NSCLC.Methods Real time-PCR was used to detect the expressions of miR-1 9a and miR-1 9b in serum of normal people and patients with NSCLC and in cancer tissues and their corresponding non-tumor tissues. The diagnostic value of miR-1 9a and miR-1 9b for NSCLC was assessed by receiver operator characteristic (ROC) curve.Results The expression levels of serum miR-1 9a and miR-1 9b were significantly higher in patients with non-small cell lung cancer than in normal people (P <0.05 ).The expressions of miR-1 9a and miR-1 9b in NSCLC were much higher than in the adjacent normal tissues.For miR-1 9a the area under ROC curve was 0.989,and the sensitivity and specificity were 95.1 and 94.0.The area under ROC curve for miR-1 9b was 0.983,and the sensitivity and specificity were 93.4 and 94.0.Conclusion The high expressions of miR-1 9a and miR-1 9b are found in cancer tissues and serum from NSCLC patients.Therefore,they may be used as noninvasive biomarkers for diagnosis and prognosis of NSCLC.

OBJECTIVE: To investigate the activation characteristics of microglia (MG) in the rats striatum with MA-induced neurotoxicity. METHODS: Male Wistar rats were divided randomly into control group (n=24) and experimental group (n=24). The rats of experimental group were injected intraperitoneally with MA (15 mg/kg x 8 injections, at 12 hours interval). The rats of control group were administrated with saline. The tissues of striatum of two rat groups were harvested at 0.5 d, 1 d, 2 d, 3 d, 4 d, 5 d, 6 d and 7 d post initial administrations of MA or saline. The structure changes were observed by transmission electron microscopy and CD-11b immunohistochemistry. The ratio of activated MG was calculated and statistically analyzed. RESULTS: In the control group, the morphological characteristics of the MG showed that the cell bodies were small with slender processes, high electronic density nucleus, and fewer organelles known as the "fork-type". In contrast, the MG in the MA-induced neurotoxicity group displayed larger cell body, shorter cell processes or disappeared, lower electronic density nucleus and rich organelles, resembling "bush-like" or "amoeba-like". The ratio of activated MG in control group was below 0.15 at all timepoints, whereas in the experimental group, the ratio of activated MG increased significantly from day 1 to day 7 (P<0.001). CONCLUSION The continuous MA stimulation of the CNS results in prominent MG activation.
Animals ; Corpus Striatum/pathology* ; Immunohistochemistry ; Male ; Methamphetamine/toxicity* ; Microglia/ultrastructure* ; Microscopy, Electron, Scanning ; Random Allocation ; Rats ; Rats, Wistar ; Staining and Labeling ; Time Factors

OBJECTIVE Cloves(Syzygium aromaticum L.)have been used as both a spice and a traditional Chinese medicinal herb for thousands of years. However, relatively little is known about its potential anticancer activity and mechanisms.In this study,we investigated the in vitro and in vivo anti-tumor effects and mechanisms of water extract of cloves(WEC)against colorectal cancer. METHODS MTS assay and Colony-formation assay were used to detect the anti-tumor activity of WEC on HT-29 cells.The in vivo anti-tumor effect of WEC was detected in a subcutaneous transplantation tumor model of human HT-29 cells.Autophagy was detected by flow cytometry and the expressions of autophagy related proteins(Beclin-1 and LC-3a/b)were determined by western blot. RESULTS MTS result showed that WEC significantly inhibited the viability of HT-29 cells,with the IC50values of 150 μg·mL-1.The colony-formation assay showed that the WEC significantly suppressed colon cancer cells proliferation.WEC also exhibited significant antitumor activity in tumor bearing nude mice. Flow cytometry result showed that WEC significantly induced autophagy, and the averaged relative values of fluorescence intensity were 206,251,341 and 356 in cells treated with 0,100,150 and 200 μg·mL-1WEC for 48 h.Western blot result showed that WEC treatment significantly increased Beclin-1 expression and ratios of LC3-II/LC3-I. CONCLUSION These result showed that WEC inhibited the growth of colon tumor both in vitro and in vivo, which might be related with autophagy induction, and WEC has potential to be developed as a novel anticancer agent for the treatment of colon cancer.