Background The pathogenesiof age-related maculadegeneration (AMD) iassociated with the senility of human retinal pigmenepithelium (RPE) cells.Seeking drug to arresRPE cell senility iof significance fothe prevention and treatmenof AMD.Research showed thathe lycium barbarum polysaccharide (LBP) can delay senility,buitinfluence on RPE cell aging iunclear.Objective Thistudy wato discusthe protective effecand mechanism of LBP on RPE cell aging.MethodPorcine retinal neural epithelial layewaisolated,and photoreceptooutesegmen(POS) waextracted by density gradiencentrifugation and marked by FITC.The POwathen co-cultured with RPE cellin the medium containing 0.01,0.10 and 1.00 g/L LBP fo24 hours.The areof fluorescence,representing the amounof POphagocytosed by RPE cells,wameasured undethe fluorescenmicroscope to evaluate the influence of LBP on the phagocytifunction of RPE cells.The POS-induced RPE lipofuscin-uptake cell model waestablished by co-culturing human RPE cellwith porcine POfo3 weeks.The RPE-POco-culture cell model waincubated in medium containing 0.01,0.10 o1.00 g/L LBP,and the autofluorescence caused by lipofuscin up-taken into RPE cellwadetected with flow cytometry.cell counting kiwaused to assescell proliferation and viability (value) 24,48 and 72 hourafteculturing.ResultPorcine POpresented athin rodundethe lighmicroscope and appeared abilayedisc-like structureundethe transmission electron microscope,and itFITC-labeled yellow-green autofluorescence waobserved undethe fluorescenmicroscope.No POwaup-taken into the RPE cellin the normal control group,buthe areof POphagocytosed by RPE cellwagradually enlarged with increasing doseof LBP,showing significandifference among the group(F =21.425,P =0.006).Compared with the POcontrol group,the phagocytosed areincreased avariouconcentrationof LBP+POgroup(P＜0.01).Flow cytometry showed thathe autofluorescence value in the POcontrol group wamore highethan thaof the normal control group.Athe LBP dose increased,the autofluorescence value in the RPE celldeclined gradually and iwaneathe normal value in the 1.00 g/L LBP+ POgroup.The rate of proliferation of the lipofuscin RPE cellvaried with the increase of doseof LBP with the maximal value in the normal RPE group and minimal value in the lipofuscin RPE group,and the rate of proliferation of the lipofuscin RPE cellascended with increasing doseof LBP until neathe normal value in the 1.00 g/L LBP + lipofuscin RPE cellgroup (P＞0.05).ConclusionLBP enhance the anti-aging effecof human RPE cellby strengthening the phagocytiability to POand the ability to remove lipofuscin and by heightening the proliferation of human RPE cells.

Abstract?AIM: To investigate the influence of propylene glycol mannite sulfate ( PGMS ) on the expression of tumor necrosis factor -α( TNF-α) and interleukin-1β( IL-1β) , in diabetic retinopathy by a rat model, to study the mechanism of PGMS against diabetic retinopathy, and provide a reliable theoretical and experimental evidence for the PGMS to be applied to clinical prevention and treatment of diabetic retinopathy.?METHODS: Male Wistar rats were randomized into 4 groups, normal control group, diabetic control group and PGMS in group, the PGMS in groups included the doses of 50mg/kg and 100mg/kg. 1% streptozotocin ( STZ) of 60 mg/kg was intraperitoneally injected in rats to establish the diabetic models. The PGMS with the doses of 50mg/kg and 100mg/kg were used to gavage in different groups of models for 12wk.Twelve weeks later, the animals were sacrificed and retinas were isolated. The aqueous humors and serums were taken, expressions of TNF-αand IL-1βprotein in retinas, aqueous humors and serums were detected by enzyme-linked immunosorbent assay ( ELISA) , respectively.The location and the expression of TNF-αand IL-1βprotein in retina tissue was detected by immunohistochemistry.?RESULTS: Twelve weeks after the use of PGMS, the level of blood glucose was not changed.ELISA showed that the expression of TNF-αand IL-1βprotein in serum and retina was significantly increased in diabetic control group than in normal control group(P<0.05), but in the groups which PGMS was given reduced, lower than those in diabetes mellitus( DM) group, especially as the concentration of PGMS increased ( P<0.05 ).But the levels of aqueous humor's TNF-αand IL-1βproteins in PGMS group were not reduced.Immunohistochemistry showed that the TNF -α protein was almost not expressed in normal control group. But the TNF-αprotein was highly expressed in diabetic control group. The expression mainly located in the ganglion cell layer, the inner plexiform layer, outer plexiform layer and pigment epithelium. The TNF-αprotein was weakly expressed at the group of 50mg/kg PGMS, the TNF-αprotein was almost not expressed at the group of 100mg/kg PGMS.When the normal control group was detected, the IL-1βprotein was weakly expressed in the outer plexiform layer.But the IL-1βprotein was also highly expressed in diabetic control group.The expression mainly located in the inner plexiform layer, outer plexiform layer and pigment epithelium. The IL -1βprotein was weakly expressed at the group of 50mg/kg and 100mg/kg PGMS.?CONCLUSION:PGMS can treat the diabetic retinopathy by downregulating the expressions of TNF-αand IL-1βin early diabetic retinopathy.PGMS maybe have a good control effect on early diabetic retinopathy.

Adult ; Female ; Humans ; Lead ; toxicity ; MMPI ; Male ; Middle Aged ; Occupational Exposure ; Personality ; drug effects

Objective To observe the morphological and functional changes of different lung cells in hyperoxia environment.Methods Type Ⅱ alveolar epithelial cells (AECⅡ) and lung fibroblasts (LFs) of fetal rats with 18 days old were isolated and culturedin vitro, and divided into air group (placed in an atmospheric incubator, and culturing with oxygen volume fraction of 0.21) and hyperoxia group (placed in a high oxygen culture chamber, and culturing with oxygen volume fraction of 0.90). Morphological changes of two kinds of cells were observed under microscope. Cell migration was observed by scratch test. The levels of reactive oxygen species (ROS) and apoptosis in cells were detected by flow cytometry.Results After 8 hours of hyperoxia, the volume of AECⅡincreased and the cells were loosely arranged; the clearance of LFs cells was increased and arranged in disorder. Scratch test showed that, compared with air group, the immigration rate of AECⅡ was inhibited at 6 hours hyperoxia [migration rate: (38.67±1.15)％ vs. (58.67±2.31)％,P < 0.01], the immigration rate of LFs was promoted at 12 hours after hyperoxia [migration rate:(55.37±1.50)％ vs. (46.90±1.20)％,P < 0.01]. With the increase of hyperoxia time, intracellular ROS contents of two cells were gradually increased, which were significantly higher than those of the air group (fluorescence intensity:130.67±4.04 vs. 54.67±2.51, 85.00±2.00 vs. 60.33±1.52, bothP < 0.01). Both two kinds of cells showed apoptosis after exposure to high oxygen, the apoptosis rate of AECⅡ at 2 hour exposure were significantly higher than that of air group [(1.93±0.28)％ vs. (1.07±0.11)％,P < 0.05], the apoptosis rate of LFs at 6 hour exposure was significantly higher than that of air group [(1.66±0.09)％ vs. (1.46±0.09)％,P < 0.05].Conclusion High concentration of oxygen can cause poor growth of lung cells, reduce AEC Ⅱ migration level and increase LFs migration, and the production of intracellular ROS eventually leads to apoptosis of lung cells.

Background and purpose:Remarkable advances have been made in cancer chemotherapy by developing new anticancer drugs and therapy strategies.However,multidrug resistance in human cancers remains a major clinical challenge for cancer treatment.Attempts in several clinical studies to reverse multidrug resistance protein (MDR) by using MDR modulators have not yet generated promising results.Our aim was to explored the mechanism of reversal of multidrug resistance in human leukemia K562 cells by PI3-K inhibitor.Methods:Trypanblue dye exclusion method was used to observe the drug sensitivity and the effect of LY294002 on the drug resistance.Western blot to analyze P-gp and p-Akt phenotypes,and flow cytometer was used to measure the intracellular drug accumulation. Results:K562/D induced by DNR was cross resistant to DNR,ADR,VCR and VP16 (Resistance Index:65,52,134 and 50 respectively).DNR induced over-expressions of P-gp and p-Akt in K562/D cells;LY294002 increased the intracellular drug accumulation,and then reversed the drug resistance to DNR,ADR,VCR and VPI6 in K562/D cells(Resistance Index:23,21,63 and 29 respectively),but not in the sensitive cells (K562/S).Conclusion:The multidrug resistance of K562/D cells can be induced by DNR which is related to the P-gp and p-Akt over-expressions, and LY294002 can reverse multidrug resistance in human leukemia cells in vitro via inhibits PI3-K/Akt pathway.

Objective To investigate whether the variability in the NS5A region among patients with hepatitis C virus(HCV) genotype 1 b affect the response to pegylated interferon alpha-2a(PEG-IFNα-2a) and ribavirin(RBV) combination therapy.Methods We retrospectively analyzed 15 patients (7 with rapid virological response and 8 non-response to therapy) with chronic HCV genotype-1b infection treated with PEG-IFNα-2a plus RBV.The entire NS5A region was amplified by reverse transcription (RT)-PCR followed by cloned sequenced.Results No single amino acid and motif substitution were associated with different responses to therapy in any of the NSSA regions.No significant differences were found in the number of mutations in the full-length NS5A,ISDR,PKRBD,and IRRDR.The number of mutations in the V3 region (aa 2356-2379)was higher in patients with rapid virological response than in patients with non-response (5.3 ± 0.5 and 3.4 ± 0.6 respectively,P =0.03).Conclusion The number of amino acid mutations in the V3 region was correlated with RVR to PEG-IFNα-2a plus RBV therapy in HCV-1b patients.

Antibiotic-associated diarrhea(AAD)in children is a type of diarrhea that occurs after the use of antibiotics in children,and its pathogenesis is closely related to the intestinal flora.The medication of antibiotics can affect the metabolic function of the intestinal flora and the immune function of the body,and then leads to the occurrence of AAD.In the view of Chinese medicine,AAD in children is mainly involved the spleen,and the etiology of the disease is due to the weakness of the spleen and stomach of the body constitution together with the attack of the pestilential pathogen and the accumulation of drug toxin.The pathogenesis of ADD in children is characterized by spleen deficiency with predominant dampness,deficiency of spleen qi,and insufficiency of spleen yang.Spleen deficiency is the root cause of pediatric AAD,and spleen and intestinal flora have commonality,so the treatment of pediatric AAD can be performed from the perspective of the spleen.The treatment of pediatric ADD from the spleen follows the principle of strengthening and activating the spleen,and the regulation of the spleen for achieving the purpose of treating the disease from the root can be achieved by the methods of strengthening spleen and draining dampness,strengthening spleen and replenishing qi,and strengthening spleen and warming yang separately with the fundamental prescriptions of Shenlin Baizhu Powder,Sijunzi Decoction,and Fuzi Lizhong Pills.

OBJECTIVETo study the relation between single nucleotide polymorphism(SNP) at the 91T-->A(Phe31Ile) position of the STK15 gene and the susceptibility of esophageal squamous cell carcinoma (ESCC) in She county--a ESCC high incidence region in North China.

METHODSPolymerase-chain reaction(PCR)-restriction fragment length polymorphism (RFLP) analysis was used to detect the genotypes of STKl5 Phe31Ile(91T-->A) SNP, and the samples came from 296 ESCC patients and 302 healthy controls.

RESULTSThe risk of ESCC significantly increased in the group which had been smoking or having a family history of upper gastrointestinal cancer (UGIC) (the OR = 1.68 and 1.77, 95% CI: 1.34-2.10 and 1.44-2.19, respectively). Rates of the three genotypes (Phe/Phe, Phe/Ile, Ile/Ile) of the STK15 Phe31Ile (91T-->A) SNPs in ESCC patients were 11.5%, 34.8% and 53.7%, respectively, and were not significantly different from that in the healthy group (11.9%, 36.8% and 51.3%) (chi2 = 0.35, P = 0.84). When compared to Phe/Phe genotype, Phe/Ile and Ile/Ile of STK15 91T-->A(Phe31Ile)did not show effect on the risk of ESCC according to the odds ratio results which were 0.98 (95% CI: 0.57-1.69) and 1.09 (0.65-1.82) respectively. STK15 91T-->A (Phe31Ile) SNP also did not significantly influence on the development of ESCC even the samples were stratified by sex, smoking status and family history of upper gastrointestinal cancer.

CONCLUSIONThe STK15 Phe31Ile(91T-->A) polymorphisms seemed irrelevant with the risk of ESCC in She county.

Aurora Kinase A ; Aurora Kinases ; Carcinoma, Squamous Cell ; genetics ; Case-Control Studies ; Esophageal Neoplasms ; genetics ; Genetic Predisposition to Disease ; Genotype ; Humans ; Polymorphism, Single Nucleotide ; Protein-Serine-Threonine Kinases ; genetics

BACKGROUND AND OBJECTIVEc-Cbl and Cbl-b are two ubiquitous members of the Casitas B-lineage lymphoma (Cbl) family, which play important roles in the downregulation of epidermal growth factor receptor (EGFR) by acting as E3 ubiquitin ligases and multiadaptor proteins. This study investigated the expression of c-Cbl, Cbl-b, and EGFR in gastric carcinoma and its clinical significance.

METHODSThe expressions of c-Cbl, Cbl-b, and EGFR were detected by immunohistochemistry using tissue microarrays consisting of 124 specimens of gastric carcinoma and 16 specimens of normal gastric mucosa. The relationship between the expressions of c-Cbl, Cbl-b, and EGFR and clinicopathologic factors of gastric carcinoma were analyzed statistically.

RESULTSThe positive rates of c-Cbl, Cbl-b, and EGFR were higher in the gastric carcinoma group than in the normal group (71.0% vs. 18.0%, P<0.01; 82.3% vs. 25.0%, P<0.01; 56.5% vs. 12.5%, P<0.01, respectively). The expression of c-Cbl was positively correlated with depth of invasion (r=0.219, P=0.015), and TNM staging (r=0.266, P=0.003). The expression of Cbl-b was positively correlated with lymph node metastasis (r=0.190, P<0.034) and TNM staging (r=0.298, P<0.001). The expression of EGFR was positively correlated with depth of invasion (r=0.286, P<0.001) and TNM staging (r=0.362, P=0.000). The expression of both c-Cbl and Cbl-b was positively correlated with EGFR (r=0.241, P=0.007; r=0.183, P=0.042, respectively). Synchronous strong-positive expressions of c-Cbl, Cbl-b, and EGFR were observed in 27 specimens of gastric carcinoma, most of which were at advanced stage.

CONCLUSIONSOverexpressions of c-Cbl, Cbl-b, and EGFR are closely related to the invasion and progression of gastric carcinoma. c-Cbl and Cbl-b may serve as novel molecular markers for gastric carcinoma.

Adaptor Proteins, Signal Transducing ; metabolism ; Adenocarcinoma ; metabolism ; pathology ; Adult ; Aged ; Aged, 80 and over ; Biomarkers, Tumor ; metabolism ; Disease Progression ; Female ; Humans ; Lymphatic Metastasis ; Male ; Middle Aged ; Neoplasm Invasiveness ; Neoplasm Staging ; Proto-Oncogene Proteins c-cbl ; metabolism ; Receptor, Epidermal Growth Factor ; metabolism ; Stomach Neoplasms ; metabolism ; pathology ; Young Adult

OBJECTIVETo summary the application of the extended free lateral arm flaps for buccal soft tissue reconstruction after buccal cancer.

METHODSFrom January to August 2011, three patients underwent the operation of buccal defect reconstruction using the extended free lateral arm flap in one-stage. PRCA was identification with the Doppler probe. According to the mark of PRCA, size and shape of defects, the flaps were designed and extended to the lateral epicondyle of humerus. The flap size ranged from 9 cm x 5 cm to 10 cm x 6 cm with a pedicle of 10 cm in length. The wounds at donor sites were closed directly.

RESULTSVascular crisis happened in one case due to local negative pressure, which resolved after emergency management. All the flaps survived completely. The patients were followed up for 6 to 10 months with no recurrence. Both the esthetic and functional results were satisfactory. Two cases suffered from numb feeling in donor sites which alleviated six months later.

CONCLUSIONSThe extended free lateral arm flap has reliable blood supply with appropriate thickness. It is an optional method for reconstruction of buccal defects after ablation of buccal cancer.

Aged ; Arm ; surgery ; Cheek ; surgery ; Facial Neoplasms ; pathology ; surgery ; Humans ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Skin Transplantation ; methods ; Surgical Flaps