The marine biological source of mineral drugs recorded in Chinese Pharmacopoeia (2010 version) mainly including pearl, nacre, clam shell, common oyster shell, ark shell, cuttle bone, and sea-ear shell are widely used in clinical. Calcium carbonate and a small amount of protein are the main components in this type of drugs. In this paper, a systematical and comparable study were carried out by determination of calcium carbonate by EDTA titration method, the crystal of calcium carbonate by X-Ray powder diffraction and the total amino acids (TAAs) of the hydrolyzed samples by ultraviolet spectrophotometry method. As a result, the crystal structure is calcite for common oyster shell, mixture of calcite and aragonite for nacre and sea-ear shell, aragonite for the other drugs. The content of calcium carbonate ranged from 86% to 96%. Cuttle bone has the highest amount of TAAs among the seven drugs which reached 1.7% while clam shell has the lowest content of 0.16% on average. In conclusion, an effective method was developed for the quality control of marine mineral drugs by comprehensive analysis of calcium carbonate and TAAs in the seven marine mineral drugs.
Amino Acids ; analysis ; chemistry ; Animal Shells ; chemistry ; Animals ; Calcium Carbonate ; analysis ; chemistry ; Crystallization ; Edetic Acid ; chemistry ; Mollusca ; chemistry ; classification ; Pharmaceutical Preparations ; analysis ; chemistry ; standards ; Quality Control ; Reproducibility of Results ; Seawater ; Species Specificity ; Spectrophotometry, Ultraviolet ; X-Ray Diffraction

Objective To investigate the expression and distribution of Diacylglycerol Kinase α (DGKα) mRNA in human hepatocellular carcinoma (HCC),and to explore the function of DGKα in the metastasis of hepatocellular carcinoma.Methods Tissues from 30 cases of HCC and 5 normal liver tissues were collected immediately after surgical resection.Semi-quantitative RT-PCR and in situ hybridization were used to detect the expression levels and distribution of DGKα mRNA,respectively.Results Semi-quantitative RT-PCR showed that the expression level of DGKα mRNA in HCC (0.798±0.317) and normal tissues (0.908±0.425) was significantly higher than those in carcinoma adjacent tissue with cirrhosis (0.205±0.102,P<0.05).In situ hybridization demonstrated that the number of DGKα mRNA positive hepatocytes in HCC [(57.6±6.3)/mm2] and normal tissues [69.8±8.7)/mm2] was significantly higher than those in carcinoma adjacent tissue with cirrhosis [(26.3±4.9)/mm2,P<0.05]; DGKα mRNA was expressed in the cytoplasm of hepatocytes in HCC and carcinoma adjacent tissue with cirrhosis,and in the nuclear of hepatocytes in normal tissues.Conclusion The present study suggests that DGKa may play important roles in carcinogenesis and progressing of HCC.

Objective To explore the mechanism underlying the rapid shortening of outflow tract and the formation of the right ventricle of the embryonic mouse heart .Methods Serial sections of embryonic mouse hearts from embryonic day 9 (E9) to E12(3 to 5 embryos for each stage)were stained with antibodies against α-sarcomeric actin (SCA), α-smooth muscle actin (SMA), GATA-4, myosin heavy chain (MHC), proliferating cell nuclear antigen (PCNA) or active caspase-3 (CAS-3).Results At E11, the aortic sac and the distal border of cardiac outflow tract had regressed towards the ventricle into the pericardial cavity , while GATA-4、SCA and SMA staining showed that precursors from the second heart field were differentiating into cardiomyocytes adding to the arterial pole of the heart to lengthen the outflow tract .The length of outflow tract rapidly shortened at E12.Before and during its shortening , no CAS-3 positive cell was detected in the entire outflow tract.During E10-12, the cardiomyocytes in the right ventricle and proximal outflow tract wall proliferated inward to form trabeculae, with some trabeculae extending into the ridges .Proximal extremities of the outflow tract ridges were gradually myocardialized remodeling into the trabeullar right ventricle wall .At E12, scattered SCA and SMA staining cells and SCA and SMA weak positive mesenchymal cell clusters , which were continuous with the outflow tract myocardium were detected in the mesenchymal proximal outflow tract ridges .These results suggested that the proximal outflow tract was remodeled into the right ventricle by trabecularization , during which mesenchymal ridges were trabecularlly myocardialized . Conclusion Ventricularization of the proximal outflow tract contributes to the trabecular right ventricle and resultes in the vapid shortening of outflow tract in the mouse embryonic heart .Cardiomyocyte appoptosis and transdifferentiation are found to play a more limited contribution during this process .

Objective To investigate the early development of the sinus venosus and the cardiac conduction system (CCS) of human embryonic hearts. Methods Serial transverse sections of 29 human embryonic hearts from Carnegie stage 10 to Carnegie stage 16 (C10-C16) were stained immunohistochemically with antibodies against α-smooth muscle actin(α-SMA),α-sarcomeric actin(α-SCA) and desmin ( DES ). Results During C12 and C13, the sinus venosus formed by confluence of systematic veins at the caudal end of the pericardial cavity could be recognized in the mesenchyme of primitive transverse septum. The mesenchymal cells of the sinus venosus gradually differentiated into α-SCA positive cardiocyocytes. At C14, the sinus venosus was within the pericardial cavity due to expansion of the pericardial cavity and incorporated into the right atrium. Differentiation of DES positive conductive cardiomyocyte was initiated in the right wall of atrio-ventricular canal of C10 embryonic heart and with the development, extended towards the myocardium of the interventricular sulcus to form His bundle, left and right bundle branches as well as the ventricular trabecular myocardium. In the atium, the strong expression of DES was first detected in the dorsal wall of C11 atrium. At C13, unique myocardial band showing α-SCA, α-SMA and DES expression in the left dorsal wall of the sinus venosus were found to be continuous with the basal wall of left atium and the dorsal wall of the atrio-ventricular canal, this band might be related to the development of conduction system from sinoatrial node to atrio-ventricular canal. During C14 to C16, primary conduction pathway of atria with strong DES expression was formed that extended from sinoatrial node along venous valves, DES positive myocardium in the dorsal and ventral walls of the atria to the right atrio-ventricular canal, respectively. Conclusions The mesenchyme of the primitive transverse septum is the heart forming field of human embryos responsible for formation of sinus venosus myocardium, cardiomyocytes are differentiated from mesenchymal cells in the primitive transverse septum and progressively added to the venous pole of the heart tube to form myocardial sinus venosus. The differentiation of CCS of the early human embryo initiates in the atrio-ventricular canal and develops gradually towards the arterial and venous poles of the heart tube. By C16, DES positive embryonic CCS can be clearly recognized morphologically.

Objective To investigate the origin of α-SMA positive cells in the outflow tract ridge of the embyonic mouse heart and to explore the ultrastructure change of the mesenchymal cells during the ridges fusion. Methods Sections of embryonic day 10(E10d) to E14d mouse embryonic hearts were stained by immunohistochemistry assay with monoclonal antibodies against α-smooth muscle actin (α-SMA), α-sarcomeric actin(α-SCA) and in situ hybridization method with PlexinA2 probe. The outflow tract ridges fusion was observed by transmission electron microscopy at E12.5d. Results From E10d to E11d, PlexinA2 positive cells were seen in the neural tube with mesenchymes around it, the aortic sac and aortic arch. These cells also migrated into the outflow tract ridge along the aortic sac wall and part of them expressed α-SMA. At E12d, PlexinA2 was expressed in the spinal ganglia, the pharyngeal mesenchyme, the aorto-pulmonary septum and the ascending aorta and pulmonary trunk. The septum showed α-SMA strongly positive. But only a few of α-SMA positive cells were observed in the ascending aorta and pulmonary trunk. At E12.5d, two clusters of condensed mesenchymal cells in the outflow tract ridges formed and began to express PlexinA2 weakly and α-SMA strongly. When the ridges began to fuse, the endothelial cells formed a cellular seam, which rapidly broke into pieces and disappeared and were replaced by the sparsed mesenchymal cells containing a few of microfilaments. Two clusters of condensed mesenchymal cells gradully moved to merge. It was noted that some mesenchymal cells contained plenty of microfilament bundles, mitochondria and focal contacts between them. Some mesenchymal cells only had a few of microfilaments and plasma membrane fusion was seen between them. Conclusionα-SMA positive cells in the outflow tract cushion may be derived from cardiac neural crest. The endothelial cells might participate in the fusion of the outflow tract ridges by endothelial-mesenchymal transformation. Mesenchymal cells of the condensed cell mass contain plenty of microfilament bundles and focal contacts or membrane fusion, which contribute to the ridges fusion.

In order to investigate the antioxidant properties of the polysaccharides from the brown alga Sargassum fusiforme, the crude polysaccharides from S. fusiforme (SFPS) were extracted in hot water, and the lipid peroxidation inhibition assay exhibited that SFPS possessed a potential antioxidant activity. Hence, two purely polymeric fractions, SFPS-1 and SFPS-2 were isolated by the column of DEAE (2-diethylaminoethanol)-Sepharose Fast Flow, with their molecular weights of 51.4 and 30.3 kDa determined by high performance gel permeation chromatography (HPGPC). They were preliminarily characterized using chemical analysis in combination of infrared (IR) and nuclear magnetic resonance (NMR) spectroscopies and found to contain large amounts of uronic acids and beta-glycosidical linkages. The antioxidant activities of these two SFPS fractions were evaluated using superoxide and hydroxyl radical-scavenging assays. The results show that the antioxidant ability of SFPS-2 was higher than that of SFPS-1, probably correlating with the molecular weight and uronic acid content.
Antioxidants ; chemistry ; Hydrogen-Ion Concentration ; Molecular Weight ; Pilot Projects ; Polysaccharides ; chemistry ; Sargassum ; metabolism

Background Our previous study demonstrated that curcumin can induce the apoptosis of retinal pigment epithelial (RPE) cells and herein inhibit the proliferation of RPE cells,and it is proved that the intravitreous injection of 0.1mg curcumin has less adverse effect to ocular tissue, inferring a good applicative prospect in clinic. Objective The goal of this experiment was to evaluate the effectiveness of curcumin on the prevention and treatment of experimental proliferative vitreoretinopathy (PVR). Methods PVR models were induced by injection of 0.1ml RPE cells (containing 2×106 cells) into vitreous cavity in 40 eyes of 20 healthy and mature New Zealand albino rabbits.0. 1ml curcumin(0. 1 mg) was then injected into lateral eye of each model rabbit immediately following the injection of RPE cells,and the equal volume of normal saline solution containing 0. 5‰ DMSO was injected into the fellow eye of each model rabbit as controls. On 1,3,7,14,21 and 28 days after injection, the changes of cornea, aqueous humor, lens, vitreous and fundus were examined and recorded by slit lamp biomicroscope, indirect ophthalmoscope,fundus color camera and B-type ultrasonograph to evaluate the inflammatory response. The incidence rate of retinal detachment was calculated and compared between curcumin group and control group. Results The inflammatory reaction in anterior chamber and misty opacity in vitreous were found from 1 day through 3 days after injection, but no obvious proliferative strap and retinal detachment in all of the experimental eyes. On the 7th day after injection, inflammatory reaction was extinct in the anterior chamber of rabbit eyes, and proliferative strap occurred in 14 eyes(75% ) in the control group but only 2 eyes (10% ) in curcumin group,showing significant difference between these two groups (P<0. 01). No retinal detachment was seen in both the two groups. On 14,21 and 28 days after injection, the incidence rate of retinal detachment was 55% ,80% ,95% respectively in control group and that of curcumin group was 10% ,15% ,15% respectively,presenting considerably differences between two groups (P<0. 01, P<0. 01 ,P<0. 01 ). Conclusion Injection of curcumin into vitreous cavity can effectively inhibit the occurrence and development of PVR in rabbit.

To construct knockout vectors containing ampicillin resistant gene and partial sequence of hyaluronidase gene(Hyl)so that Hyl can be knock out by transforming the plasmid into Streptococcus zoopidemics mutans.First,partial sequence of Hyl(Hyl-1)was cloned into the vector of pMD19-T by using DNA of Streptococcus zoopidemics as template,and then a knockout vector pMD19T-SA was constructed,in which Hyl-1 gene was disrupted by inserting ampicillin resistant gene(Amp)from reverse PCR.As expected,the vector was proved to be consisted of Hyl-1-Amp-Hyl-1-pMD19-T.Thereafter,DNA fragment of Hyl-1-Amp-Hyl-1 was subcloned into pBR322 vector,the resulting construct was then checked by PCR and restriction analysis for the proper configuration of the knockout vector pBR322-SA.Both of the knockout vectors were used to transform Streptococcus zoopidemics and one recombinant was obtained in result.From results of PCR and Hyl activity assay,it was indicated that in the recombinant the Hyl gene was disrupted completely.

Objective To study the mechanism of marcosomia by investigating insulin-like growth factor 2(IGF_2)imprinting status,expression level and the promoter usage in the placenta of macrosomia. Methods We selected heterozygous cases for Apa Ⅰ polymorphism in exon 9 of IGF_2 gene and then analyzed its imprinting status in 168 placentas of macrosomia and normal pregnancies.IGF_2 transcription levels and promoter usages in macrosomic and normal placenta were evaluated by using semi-quantitative RT- PCR assay.Results Thirty specimens of macrosomic placenta and 30 of normal placenta were identified as heterozygous for IGF_2.All of the heterozygous specimens showed maintenance of imprinting.The expression of placental IGF_2 mRNA(2.2?1.2)was significantly higher in macrosomia than that of normal weight group (1.6?0.6,P 0.05).Conclusion It is possible that over expression of IGF_2 in placenta contributes to macrosomia while the promoter usage and imprinting status are not associated with macrosomia.

OBJECTIVETo study the expression level and significance of glucose transporter 1 (Glut-1) in normal breast tissue, adenosis, adenoma and breast carcinoma.

METHODSA total of 147 cases of female breast tissue samples, including 92 cases of invasive ductal carcinoma, 26 cases of breast fibroadenoma, 24 cases of breast adenosis and 5 cases of normal breast tissues, were collected for quantitative detection of the expression of Glut-1 protein by immunohistochemistry (EnVision method) and Western blot, and its mRNA by reverse transcriptase-polymerase chain reaction (RT-PCR).

RESULTSIn normal breast tissue and benign lesions of the breast, Glut-1 was undetectable or only weakly detectable in cytoplasm of ductal and acinar epithelia. In contrast, the intensity of Glut-1 staining was significantly higher in invasive ductal carcinomas (P = 0.0002) with protein expression predominantly in cellular membrane and lesser in cytoplasm. Western blot and RT-PCR analyses showed that the expression of Glut-1 protein and mRNA were significantly increased in invasive ductal carcinoma than fibroadenoma (P =0.001 for protein; P <0.05 for mRNA) and adenosis (P =0.001 for protein; P < 0.05 for mRNA). There was a significant difference among groups (P = 0.0002 for protein; P = 0.0001 for mRNA).

CONCLUSIONSGlucose transport activity, as indicated by Glut-1 protein and its mRNA expression, significantly increases in breast carcinoma than non-cancerous lesions. The over-expression of Glut-1 in breast carcinoma is tightly coupled with tumor cell proliferation, invasion and metastasis, implying that Glut-1 may serve as a new marker in the early diagnosis and prognostication of breast malignancy as well as a new therapeutic target.

Breast Neoplasms ; metabolism ; Carcinoma, Ductal, Breast ; metabolism ; Female ; Gene Expression Regulation, Neoplastic ; Glucose ; physiology ; Glucose Transport Proteins, Facilitative ; genetics ; metabolism ; Glucose Transporter Type 1 ; genetics ; metabolism ; Humans ; Prognosis