Dissent and Disputes ; Humans ; Occupational Diseases ; diagnosis

Objective To study the expression of p53,p16,PCNA protein in esophageal carcinoma and its relationship to sexual distinction,the location of disease,the biological level,the depth of invasion and lymph node metastasis.Methods 118 patients with esophageal carcinoma were included in the study,all of them were treated for the first time.p53,p16 and PCNA protein in the 118 cases of esophageal carcinoma were detected by immunohistochemical assay(SP technique). Results The positive expression of p53, p16, PCNA protein in 118 patients was 80 %(92/118),42%(50/118)and 97%(115/118),respectively.The positive expression of p53,PCNA protein were irrelated to the sexual distinction,the location of disease,the biological level,the depth of invasion and the lymph node metastasis.The loss of p16 was significantly related to the depth of invasion and the lymph node metastasis(P

Background Growth factor-induced proliferation and migration of retinal pigment epithelium (RPE) cells are the major pathological changes of proliferative vitreoretinopathy (PVR).Arsenic trioxide ( As2O3 ) is an active ingredient of Chinese traditional medicines,which has an inhibition on proliferation and migration of tumor cells.However,it is not clear whether As2O3 could inhibit growth factor-induced proliferation and migration of RPE cells. Objective This study was to explore the effects of As2O3 on epidermal growth factor (EGF)-induced proliferation and migration of ARPE-19 cells. Methods RPE cell line (ARPE-19 cells) were cultured.Different concentrations of As2O3(0,0.5,1.0,2.0,5.0,10.0,20.0 μmol/L) were added in the culture plate to treat ARPE-19 cells with or without 10 mg/L EGF in serum-free group for 24 and 48 hours,respectively.The MTT colorimetric assay was used to check the cell viability and evaluate the drug toxicity.The effects of As2O3 on EGF-induced proliferation of ARPE-19 cells were analyzed to get an effective and avirulent concentrations of As2O3.The effects of As2O3 on EGF-induced migration of ARPE-19 cells were observed by scratch-wound assay and the Boyden chamber assay.Results MTT assay showed that the A values were gradually declined with the increase of As2O3 concentrations after As2O3 treatment without EGF for 24 hours and 48 hours ( Fgroup =38.269,P =0.000 ; Ftime =0.874,P =0.358 ).Compared with the control group,no significant differences were seen in the A values of ARPE-19 cells in 0.5-5.0 μmol/L groups (all P＞0.05).Meantime,As2O3 reduced the A values of ARPE-19 cell with 10 mg/L EGF in dose- and time-dependent manner ( Fgroup =152.155,P =0.000 ; Ftime =51.649,P =0.000 ).There were not significant differences in 10 mg/L EGF-induced cell growth after 0.5,1.0,2.0 μmol/L As2O3 was added for 24 and 48 hours ( Fgroup =2.215,P =0.126 ;Ftime =2.230,P =0.155).However,when 5.0-20.0 μmol/L As2O3 added,the A values of 10 mg/L EGF-induced ARPE-19 cells lowed,showing a significant difference in comparison with the control groups ( all P＜0.05),with the cellular inhibiting rate 12％,32％,37％ in 24 hours and 39％,44％ and 53％ in 48 hours.Scratch-wound assay showed that EGF-induced horizontal migration of ARPE-19 cells was slow after 0.5-2.0 μmol/L As2O3 treated,and the same results also appeared in cell lognitudinal migration by Boyden chamber assay,with the inhibitory rates 22％,33％ and 46％ respectively. Conclusions As2O3 is avirulent on ARPE-19 cells within definite concentration range.At ≤ 2.0 μmol/L concentrations,As2O3 dose not affect EGF-induced proliferation of ARPE-19 cells,but it suppresses EGF-induced cell migration.At ≥ 5.0 μmol/L concentrations,As2O3 plays an inhibitory role to EGF-induced proliferation of ARPE-19 cells.

OBJECTIVEThe objective of this study was to observe the interventional effect of cod liver oil supplementation on re-vaccination to hepatitis B virus (HBV) among infants and young children.

METHODSAll 7-36 months old infants and young children, who had been vaccinated with obligatory HBV vaccines routinely by the national technical and administrative procedures for HBV vaccination on children of China, were convened among villages in Linyi, Shandong province, from October 2008 to March 2009. After detection of serum anti-HBV, one hundred children with lower serum anti-HBV were picked out for the randomized, double blinded, placebo controlled vitamin A supplementation study. The children in the intervention group (50 subjects) took 0.5 g condensed cod liver oil (containing 25 000 IU vitamin A and 2500 IU vitamin D(2)) every 15 days for six times. The children in the control group (50 subjects) were given corn oil with same volume. All children were re-vaccinated at the 30th and the 60th day of the experiment. The serum samples were collected from each child at the 90th day of the experiment. Retinol concentration in serum samples was analyzed with HPLC method before and after the intervention. The levels of serum anti-HBs were detected by the electro-chemi-luminescence immunoassay (ECLIA).

RESULTSTotal 74 children finished the supplemental experiment and blood collection, 37 subjects in each group, respectively. After intervention, the serum retinol level in the experimental and control group were (404.1 ± 123.1) and (240.8 ± 92.8) µg/L (t = 6.441, P < 0.01), respectively. The serum anti-HBs levels in the experimental and control group were (2737.2 ± 2492.6) and (1199.7 ± 2141.6) U/L (t = 2.846, P < 0.01), respectively. The rate of weak or no-answer case in experimental and control groups was 0.00% (0/37) and 10.81% (4/37) (χ(2) = 4.229, P = 0.040), respectively.

CONCLUSIONThe results showed that vitamin A supplementation might enhance the re-vaccination reaction against HB vaccine in infants and young children.

Child, Preschool ; Cod Liver Oil ; therapeutic use ; Dietary Supplements ; Double-Blind Method ; Hepatitis B ; prevention & control ; Hepatitis B Antibodies ; blood ; immunology ; Hepatitis B Vaccines ; immunology ; Hepatitis B virus ; immunology ; Humans ; Immunity, Active ; Infant ; Vitamin A ; therapeutic use ; Vitamins ; therapeutic use

OBJECTIVETo observe the influence of Pilose antler polypeptide on the glycosaminoglycan and type II collagen in the articular cartilage in experimental knee osteoarthritis.

METHODSTotally 64 New Zealand white rabbits of 6 months old were randomly divided into 2 groups:normal group (n = 8) and model group (n = 56). Model group was surgically induced into osteoarthritis model by method of Hulth. After successful modeling, the rabbits of model group were further divided into 2 groups: Pilose antler polypeptide-treatment group and control group, 24 rabbits in each group. Pilose antler polypeptide-treatment group received 0.5 ml intra-articular injection of Pilose antler polypeptide dilution liquid once in per 2 days for 30 days, while control group received 0.5 ml intra-articular injection of physiological saline. On days 7, 15 and 30 after intervention, articular cartilage samples were collected respectively. The content of glycosaminoglycan in articular cartilage was observed by toluidine blue staining and the expression of type II collagen in cartilage matrix was detected by immunohistochemical staining.

RESULTSAlong with the prolonging of time, the content of glycosaminoglycan and type II collagen in cartilage matrix of the Pilose antler polypeptide-treatment group and control group decreased gradually. On days 7, 15 and 30 after intervention, integrated optical density of the type II collagen positive area in cartilage matrix of the Pilose antler polypeptide-treatment group were (312.06 +/- 14.12), (273.31 +/- 12.42) and (248.34 +/- 10.41), which had statistically significant differences. Integrated optical density of the type II collagen positive area in cartilage matrix of the control group were (253.47 +/- 15.53), (215.67 +/- 9.72) and (160.01 +/- 13.23), which had statistically significant differences. At the same period, integrated optical density of the type II collagen positive area in cartilage matrix of the Pilose antler polypeptide-treatment group was higher than that of control group, which had statistically significant difference.

CONCLUSIONPilose antler polypeptide can inhibit reduction of the glycosaminoglycan and type II collagen in cartilage matrix and delay the degeneration of articular cartilage.

Animals ; Antlers ; chemistry ; metabolism ; Collagen Type II ; metabolism ; Disease Models, Animal ; Female ; Glycosaminoglycans ; metabolism ; Humans ; Male ; Osteoarthritis, Knee ; drug therapy ; metabolism ; Peptides ; metabolism ; pharmacology ; Rabbits

BackgroundThe quest to look for seed cells is a hot spot of cornea transplant research in solving the problem of the lack of donor. Bone marrow mesenchymal stem cells(BMSCs) have been successfully induced into retinal ganglion cells(RGCs) in vivo,but the successful induction of BMSCs into corneal endothelial cells has not been reported.Objective This experiment was to study the transplantation of BMSCs on corneal endothelial surface using the splitting Descemet's membrane. MethodsFour healthy adult rhesus monkeys were divided into the experimental group ( 3 monkeys) and control group ( 1 monkey). Mesenchymal stem cells (MSCs) were isolated from bone marrow by density gradient centrifugation combined with adhering means. The cultured cells were identified by flow cytometry and its ability to differentiate was determined by allowing them to differentiate into adipocytes in vitro and labeled by 5-bromodeoxyuridine ( BrdU ) for subsequent identification. Corneal grafts of 7 mm in size with tearing of the Descemet' s membrane were prepared in the experimental group and control group. After labeling by 5-bromodeoxyuridine( BrdU ) ,cultured cells were transplanted onto the endothelial surface of cornea grafts in the experimental group, but no cultured cells were seeded in the graft of the control group. The corneal grafts were then sutured in situ, and were removed 1,2 or 3 months after operation to examine the distribution and connection between transplanted cells and their morphologic changes under the electron microscope. Results High purity MSCs were harvested by density gradient centrifugation combined with adhering method. Cultured cells reached confluency after 12 to 16 days, presenting with a spindle shape and parallel or swirling arrangement. Flow cytometry analysis showed that 94.26％ of cells were positive for CD29,7. 51％ for CD34 and 4. 02％ for CD45. Larger nuclei filled with plastosomes, golgiosomes and rough endoplasmic reticula were found on the graft under the transmission electron microscope( TEM ). After 3 weeks, MSCs were differentiated into adipocytes where Oil Red O staining resulted in an orange-red staining in the cytoplasm and blue staining in the nuclei. The transplanted cells attached loosely on the endothelial surface of the corneal graft and came in contact with each other in one month. The shape of the cells appeared as spindle-shaped and polygonal after 2 months and became tightly packed after 3 months. The positive cells retained the BrdU label and presented with brown nuclei. No endothelia cells grew in the cornea graft in the control group, with an absence of BrdU labeling. Conclusions Mesenchymal stem cells can be transplanted onto the corneal endothelial surface successfully and form a monolayer using the centrifugation method, and present with good survival and proliferation ability.

Anxiety ; etiology ; Asthma ; psychology ; Child ; Child, Preschool ; Depression ; etiology ; Female ; Humans ; Male ; Mental Disorders ; etiology

OBJECTIVETo obtain a simple and effective method to isolate and purify adult Leydig cells.

METHODSThe testes of human adults were digested and then the density gradient centrifugation of the cells was performed with four different Percoll concentrations (60%, 34%, 26%, 21%) to isolate Leydig cells, whose characteristics were identified by cytological observation staining, 3beta-HSD staining and detection of hCG and testosterone secretion.

RESULTSHigh-concentration (> 90%) purified Leydig cells were acquired, and many identification experiments demonstrated the adequate testosterone secretory function of the isolated and purified Leydig cells.

CONCLUSIONThis method is easy and efficient for the isolation and purification of adult Leydig cells.

Adult ; Cell Separation ; methods ; Cells, Cultured ; Chorionic Gonadotropin ; pharmacology ; Humans ; Leydig Cells ; cytology ; drug effects ; secretion ; Male ; Povidone ; Silicon Dioxide ; Testosterone ; secretion

In order to establish HPLC fingerprint of Liuwei Dihuang soft capsule, and to provide certain reference for an quality control of it, the HPLC method was performed on an Agilent C18 (4.6 mm x 250 mm, 5 μm) column with acetonitrile-0.02% trifluoroacetic acid as mobile phase, gradient elution volume flow of 1.0 mL x min(-1), column temperature was 30 degrees C, detection wavelength: 0-60 min, 238 nm, 60-70 min, 210 nm. The software for chromatographic fingerprint was applied to analysis different batches of Liuwei Dihuang soft capsule samples. Sixteen mutual peaks were selected as the fingerprint peaks in 12 samples with loganin as the reference peak, and all of the detected peaks were separated effectively. Cluster analysis (HCA) and similarity analysis (SA) were done based on data of 12 samples clustering analysis of 12 batches of samples were divided into 2 categories. Including 7 for the first class, the rest was second, similarities calculated by SA were all above 0.92, indicating a good similarity between the reference and twelve batches of samples, also, the analysis results of HCA and SA basically the same. This method is simple with good precision, repeatability and stability, and provides the basis for Liuwei Dihuang soft capsule quality control.
Capsules ; analysis ; Chromatography, High Pressure Liquid ; methods ; Drugs, Chinese Herbal ; analysis ; Quality Control

OBJECTIVETo investigate if the interaction between TGF-beta1/Smad pathway and ERK pathway in vascular smooth muscle cells exists.

METHODSThe rat arota was removed. The primary VSMC were isolated and cultured in vitro, then the VSMC were divided into four groups: (1) control group, (2) (TGF-beta1 group, (3) ERK blocking agent group, (4) TGF-beta1 + ERK blocking agent group. The expression of Smad2/3, ERK1/2 proteins, the content of phosphorylated ERK1/2 and Smad2/3 proteins were detected by Western blot, and the expression of Smad2/3 mRNA was detected by reverse transcription-polymerase chain reaction(RT-PCR) .

RESULTS(1) In contrast to control group, the content of phosphorylated Smad2/3 and phosphorylated ERK1/2 proteins in TGF-beta1 group was increased (P < 0.05), that in ERK blocking agent group was decreased (P < 0.05). There was no difference between control group and TGF-beta1 + ERK blocking agent group. Compared with TGF-beta1 group, the contents of phosphorylated Smad2/3 and phosphorylated ERK1/2 proteins in TGF-beta1 + ERK blocking agent group was decreased (P < 0.05). There was no difference in the expression of Smad2/3 and ERK1/2 proteins among different groups. (2) There were no differences in expression of Smad2 and Smad3 mRNA among different groups.

CONCLUSION(1) TGF-beta1 can induce Smad2/3 proteins to be phosphorylated dependent on the activated ERK pathway. (2) ERK pathway does not effect the expression of Smad2/3 at the level of protein and mRNA.

Animals ; Aorta ; cytology ; Cells, Cultured ; Female ; MAP Kinase Signaling System ; physiology ; Male ; Mitogen-Activated Protein Kinase 3 ; metabolism ; Muscle, Smooth, Vascular ; cytology ; physiology ; Myocytes, Smooth Muscle ; physiology ; Phosphorylation ; Rats ; Rats, Wistar ; Signal Transduction ; Smad Proteins ; metabolism ; physiology ; Transforming Growth Factor beta1 ; physiology