As pattern recognition receptors of innate immunity,Toll-like receptors (TLRs) play an important role in the identification of pathogens,regulation of adaptive immunity and signal transduction.With more exploration of the mechanism of microbial keratitis and the acute or chronic rejection after penetrating keratoplasty,the induction and regulation of TLRs on the corneal immune response is increasingly concerned.Up to now,more and more studies of immune signal pathway of TLRs in mediating inflammatory response of bacterial keratitis,fungal keratitis,viral keratitis and graft rejection have been documented.To regulate the expression of TLRs is undoubtedly available for lightening and arresting the immuno-inflammatory response.Here,the current knowledge about TLRs,their signal transduction pathway and their regulation on corneal immunity are summarized,aiming to discuss their significant effects and the potential therapeutic targets.

Objective To investigate the water quality of finished water of the centralized water supply system in Shenzhen according to 106 indexes of water quality standards of China.Methods The collection and preservation of water samples was according to the standard examination methods for drinking water-collection and preservation of water samples (GB/T 5750.2-2006).The sanitary quality of the finished water samples collected from 35 centralized water supply systems in cities were determined and evaluated according to the Standards for Drinking Water Quality (GB 5749-2006) in May of 2008.Results The average qualified rate of drinking water was 82.9% (29/35) in Shenzhen.Among 106 indexes,4 indexes (such as turbidity,aluminum,manganese and free chlorine residue) exceeded the standard limits in degrees in some centralized water supply system.Conclusion According to the results of the present paper,it is considered that the water quality of the product water from the centralized water supply system in Shenzhen is good.

Objective To determine the value of MR cholangiography(MRC)in the preoperative evaluation of biliary anatomy of living liver donors.Methods Fifty eight consecutive donors underwent MRC examinations and living liver transplantation.MRC was performed on a 1.5 T scanner with breath-hold rapid acquisition of T_2WI slab and breathing-gating 3D FSE T_2WI.Images of MRC and IOC were compared and classified according to the modified Huang's classification.Results Thity four(58.6%)liver donors showed normal biliary anatomy on IOC,and 24(41.4%)donors revealed variant bile anatomy.MRC correctly depicted biliary anatomy in 91.4%(53/58)donors.The sensitivity,specificity,positive predictive value and negative predictive value of MRC in distinguishing normal and difierent types of variant biliary anatomy were 83.3%(20/24),100%(34/34),100%(20/20),89.5%(34/38)respectively.Conclusion MRC can accurately assess the biliary anatomy in living liver donors and may guide the preoperative planning of liver transplant.

Objective To investigate the quality assurance (QA) and quality control (QC)of intravenous injection in the radiology department. Methods The data from 1352 cases of intravenous injection in the radiology department were analyzed retrospectively. Results The bulge in region of the intravenous injection have been found in 28 cases, regional cyanochroia in 52 cases, allergic response in 25 cases, hypoglycemia response in 58 cases. Conclusion It is important to strengthen the QA and QC of intravenous injection in the radiology department.

Objective:To observe the difference of results of treadmill exercise test (TET) between male smokers and non-smokers .Methods:A total of 200 men with long-term smoking ≥20 cigarette per day were screened from outpatients and treated as smoking group .Another 200 men with similar age ,who didn’ t smoke at all ,were regarded as non-smoking control group (control group) .There were no medical history for hypertension ,hyperlipidemia ,diabetes ,and were nega-tive for electrocardiography and color echocardiography (exclude left ventricular hypertrophy etc .organic heart disease) in two groups .TET results and incidence rate of acute myocardial infarction (AMI) within two years were compared between two groups .Results:Compared with control group ,there were significant rise in TET positive rate (37. 5% vs .57. 5% , P<0.001) and incidence rate of AMI (8.5% vs .17.5% ,P=0.007) within two years in smoking group .Conclusion:Risk of suffering from coronary artery disease is more high in smokers among men .

OBJECTIVETo investigate the effects of prostate stromal cells from different zones of normal prostate tissue on the growth of prostate cancer cells and their action mechanisms.

METHODSWe extracted stromal cells in the fresh normal prostatic tissue derived from the peripheral zone (PZ) or transitional zone (TZ), amplified them in vitro, and used the supernatants of the cells as conditioned media to culture hormone-resistant prostate cancer DU145 cells. We measured the growth curve of the tumor cells using the CCK8 method, determined the number and viability of the cells by trypan blue staining, evaluated their invasiveness by scratch test, and detected the effects of the stromal cells on the key enzymes in the glycolysis of the tumor cells by Western blot.

RESULTSThe conditioned medium with the PZ-derived stromal cells promoted, while that with the TZ-derived stromal cells inhibited the growth of the tumor cells. The former significantly increased, while the latter markedly decreased the expressions of the key enzymes hexokinase 2 (HK-2), pyruvate kinase 2 (PKM-2), lactate dehydrogenase (LDHA), and pyruvate dehydrogenase (PDH) in the glycolysis of the tumor cells.

CONCLUSIONProstate stromal cells from different zones exert different influences on the growth of tumor cells, which may be associated with their different effects on the glycolysis of tumor cells.

Blotting, Western ; Cell Culture Techniques ; Cell Proliferation ; Culture Media, Conditioned ; Glycolysis ; Humans ; Male ; Prostate ; cytology ; Prostatic Neoplasms ; pathology ; Stromal Cells ; physiology ; Tumor Cells, Cultured

OBJECTIVETo investigate the apoptosis and proliferation effect of matrine on human medulloblastoma cell line D341 in vitro and the effect of the expression of the related caspase 3 and caspase 9 proteins.

METHODSThe D341 cells were cultivated successfully in vitro. Then the cells were divided into 5 groups according to the concentration of matrine (0.5 mg/mI group, 1.0 mg/ml group, 1.5 mg/ml group, 2.0 mg/ml group and the control group was 0 mg/ml). All the experiments were repeated three times. The cell morphologic and structure change was observed with the optical microscope and the transmission electron microscope. The proliferation of D341 cell was analyzed using Cell Counting Kit-8 assay. Apoptosis was detected by Annexin V-FITC/PI double staining. The expression of Caspase3 and Caspase9 was detected by Western blot.

RESULTSWith the effect of matrine, the proliferation inhibition rate gradually increased with drug concentrations increasing, and there was a significant difference (P < 0.01). The inhibitory effect of matrine on cell proliferation was different with the different treatment time, there was a significant difference between the 24 h to 72 h groups (P < 0.01). The apoptotic rate increased with matrine concentrations increasing. There were significant differences between the group of 0.5 mg/mI or 1.0 mg/mI to the group of 1.5 mg/mI or 2.0 mg/mI (P < 0.05). The apoptotic rate increased with the prolonged treatment time. There were significant differences between the group of 24 h or 48 h to the group of 72 h ( P < 0.05). With the increase of matrine concentration, the expression of Caspase 3 and Caspase 9 increased (P < 0.01).

CONCLUSIONMatrine induces the apoptosis, and inhibits the proliferation of human medulloblastoma D341 cells in vitro by up-regulation of the expression level of Caspase3, Caspase9.

Alkaloids ; pharmacology ; Apoptosis ; Caspase 3 ; metabolism ; Caspase 9 ; metabolism ; Cell Line, Tumor ; Cell Proliferation ; Cerebellar Neoplasms ; metabolism ; pathology ; Humans ; Medulloblastoma ; metabolism ; pathology ; Quinolizines ; pharmacology ; Up-Regulation

Objective To investigate the sequence characteristic of black bear's mitochondrial NADH-ubiquinone oxidoreductase chain 4 gene(ND4).Methods Primers were designed according to certain species' NADH-ubiquinone oxidoreductase ND4 in which the DNA sequence had been reported.The DNA sequence was amplified by PCR from the Asian black bear Sichuan subspecies's(Ursus thibetanus mupinensis)hair-follicle tissue total DNA and were analyzed with its amino acid sequence.Results The ND4 gene sequence length was 1 402 bp and contained a 1 377-nucleotide open reading frame encoding 459 amino acid residues.The amino acid sequence's pI was 9.37 and weight was 51.5?10 3.The DNA sequence and corresponding amino acid sequence of black bear's ND4 gene had the high similarity with those of other mammalian animals' ND4 gene,and the similarity was 89% to 91% and 94% to 95%.The result of phylogenetic tree was basically consistent with the traditional species system phylogenetic relations.Topology prediction showed the Asian black bear Sichuan subspecies's ND4 proteins contained N-glycosylation site,kinase C phosphorylation sites,casein kinase Ⅱ phosphorylation site,N-myristoylation site and leucine-rich region.In addition,black bear's ND4 protein contained leucine zipper pattern and Leucine-rich region.Conclusion The mammalia animal's ND4 gene has the high conservative nature.The pretein coded by ND4 gene has high uniformity in the function.

Type 1 diabetes is a autoimmune disease that occurs under the influence of both genetic predisposition and environmental factors,mediated mainly by T lymphocytes with various kinds of other involved immune cells.The autoimmune attacks eventually lead to the islet beta cell destruction and insulin insufficiency.Multiple fundamental studies and clinical trials have revealed that umbilical cord blood pluripotent stem cells have the potential to help restore immune balance,induce immune tolerance,stop the autoimmune attacks against beta cells,and promote beta cell regeneration in type 1 diabetes mellitus (T1DM),by means of cell to cell contact and soluble cytokine secretion.For the past few years,the National Clinical Research Center for Metabolic Diseases of Second Xiangya Hospital has been leading the research on stem cell education therapy and has performed the therapy for 35 juvenile type 1 diabetes patients from China and foreign countries,introducing a novel treatment for T1DM.

Objective To study the effects of midkine (MK) gene-targeting small interfering RNA (siRNA)on the invasion of melanoma cells.Methods Three MK gene-targeting siRNAs (S1,S2 and S3)were designed,constructed,and transfected into human A375 melanoma cells.Real-time PCR was performed to measure the expression of MK gene and to screen the siRNA with best efficacy.Then,A375 cells were transfected with the optimal siRNA of various doses (3.125,6.25 and 12.5 nmol/L)followed by additional culture of various durations(24,48,72 hours).Some A375 cells remaining untreated served as the blank control group,and some transfected only with liposomes served as the vector control group.Reverse transcription (RT) -PCR and Western blot were conducted to detect the mRNA and protein expression of MK,respectively,MTT assay to observe the adhesion of A375 cells,and Boyden chamber was used to evaluate cell invasion.Results The expression of MK mRNA was downregulated by all the three siRNAs,especially by the siRNA S3,which was used in the following transfection experiment.Real-time quantitative PCR revealed that the MK mRNA expression was reduced by the siRNA in a dose- (r24hours=-0.906,r4Bhours=-0.922,r72hours=-0.939,all P<0.01)and time-dependent(r3.125nmol/L=-0.889,r625nmol/L=-0.935,r125nmol/L=-0.928,all P<0.01)manner.MTT assay showed that the percentage of adhesing cells was 73.66%±2.25%,49.36%±2.16%and 28.35%±1.68%in A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively.The number of cells migrating across the chamber filter was 23.9±1.6,12.1±1.5,5.6±1.2 among A375 cells transfected with the siRNA of 3.125,6.25 and 12.5 nmol/L,respectively,significantly lower than that in the blank control group(36.8±1.5).The percentage of adhesing cells and number of migrating cells decreased with the dose of siRNA(r=-0.936,-0.915,P<0.01,0.05,respectively).Conclusions MK gene might play an important role in the adhesion and invasion of melanonla cells.To down-regulate the expression of MK gene by siRNA may suppress the adhesion and invasion of melanoma cells.