In this study, the psbA-trnH sequence as DNA barcode was used to evaluate the accuracy and stability for identification pteridophyte medicinal material Pyrrosiae Foliumas from adulterants. Genomic DNA from 106 samples were extracted successfully. The Kimura 2-Parameter (K2P) distances and ML tree were calculated using software MEGA 6.0. The intra-specific genetic distances of 3 original plants were lower than inter-specific genetic distances of adulterants. The ML tree indicated that Pyrrosiae Folium can be distinguished from its adulterants obviously. Therefore, the psbA-trnH sequence as a barcode of the pteridophyte, can accurately and stably distinguish Pyrrosiae Folium from its adulterants.
Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Ferns ; classification ; genetics ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Quality Control

SARS coronavirus (SARS-CoV) is the etiologic agent of severe acute respiratory syndrome. The aim of this study was to construct Sars-CoV membrane (M), nucleocapsid (N) and spike 2 (S2) gene eukaryotic expression plasmids, and identify their expression in vitro. Gene fragments encoding N protein, M protein and S2 protein of SARS-CoV were amplified by PCR using cDNA obtained from lung samples of SARS patients as template, and subcloned into pcDNA3.1 vector to form eukaryotic expression plasmids. SARS-CoV protein eukaryotic expression plasmids were transfected respectively into CHO cells. Immunohistochemistry was employed to detect the expression of the structural proteins of SARS-CoV in transfected cells. SARS-CoV protein eukaryotic expression plasmids were successfully constructed by identification with digestion of restriction enzymes and sequencing. M, N and S2 proteins of SARS-CoV were detected in the cytoplasm of transfected CHO cells. It was concluded that these recombinant eukaryotic expression plasmids were constructed successfully, and SARS-CoV encoding proteins could activate transcription and expression of hfgl2 gene.

Smart dressings, which show obvious advantage and potential in wound treatment and real-time monitoring, attract widespread attention in recent years. Real-time and dynamic acquiring wound information is vital to the treatment and prognosis of wound. Further research on smart dressings is helpful for wound management, personalized treatment, and realization of medical application translation of health monitoring technology. In the article, we categorize smart dressings and conclude their functions according to the type of micro-environment information of wound gathered by smart dressings.

OBJECTIVETo study the biological effects of 585 nm pulsed dye laser (FLPDL) in the treatment of congestive scar.

METHODSBy histological study, collagen VG staining and microvascular staining, we investigated the changes of collagen fibers and the density of microvessels in the congestive scars after FLPDL treatment.

RESULTSHistological and immunohistochemistry examinations showed that FLPDL achieved normal vascularity in the scar after over 3 times of treatment.

CONCLUSIONSPDL treatment can change fundamentally the physiology of wound healing if applied in the early phases.

Adolescent ; Adult ; Cicatrix ; therapy ; Female ; Humans ; Lasers, Dye ; Low-Level Light Therapy ; methods ; Male ; Middle Aged ; Reconstructive Surgical Procedures ; methods ; Treatment Outcome ; Wound Healing ; Young Adult

Acute pancreatitis (AP) is a common acute abdomen in clinic with a rapid onset and dangerous pathogenetic condition. AP can cause an injury of intestinal mucosa barrier, leading to translocation of bacteria or endotoxin through multiple routes, bacterial translocation (BT), gut-origin endotoxaemia, and secondary infection of pancreatic tissue, and then cause systemic inflammatory response syndrome (SIRS) or multiple organ dysfunction syndrome (MODS), which are important factors influencing AP's severity and mortality. Meanwhile, the injury of intestinal mucosa barrier plays a key role in AP's process. Therefore, it is clinically important to study the relationship between the injury of intestinal mucosa barrier and AP. In addition, many factors such as microcirculation disturbance, ischemic reperfusion injury, excessive release of inflammatory mediators and apoptosis may also play important roles in the damage of intestinal mucosa barrier. In this review, we summarize studies on mechanisms of AP.
Acute Disease ; Animals ; Humans ; Intestinal Mucosa ; blood supply ; injuries ; metabolism ; Malnutrition ; pathology ; Microcirculation ; metabolism ; Pancreatitis ; complications ; metabolism ; pathology ; physiopathology ; Reperfusion Injury ; pathology

In order to evaluate the efficiency of ITS2 and psbA-trnH sequences used as DNA barcodes to distinguish Plantaginis Semen from its adulterants, we collected 71 samples of Plantaginis Semen and its adulterants. The ITS2 and psbA-trnH sequences were aligned through Clustal W, and the genetic distances were calculated by kimura 2-parameter (K2P) model and the Neighbor-Joining (NJ) phylogenetic trees were constructed using MEGA 5.1. The results indicated that the ITS2 sequence lengths of Plantago asiatica and P. depressa were 199 bp and 200 bp, respectively; the maximum intra-specific K2P distance were lower than the minimum inter-specific K2P distance; the NJ tree based on ITS2 sequence indicated that Plantaginis Semen and its adulterants could be distinguished clearly. The sequence lengths of psbA-trnH of both P. asiatica and P. depressa were 340 bp; the maximum intra-specific K2P distances were lower than the minimum inter-specific K2P distance; the NJ tree based on psbA-trnH sequence showed that Plantaginis Semen can be distinguished clearly from its adulterants except for P. major. Therefore, ITS2 sequences can be used as an ideal DNA barcode to distinguish Plantaginis Semen from its adulterants.
Base Sequence ; DNA Barcoding, Taxonomic ; methods ; DNA, Plant ; genetics ; DNA, Ribosomal Spacer ; genetics ; Drug Contamination ; prevention & control ; Drugs, Chinese Herbal ; chemistry ; classification ; Molecular Sequence Data ; Phylogeny ; Plant Proteins ; genetics ; Plantago ; classification ; genetics ; Quality Control ; Seeds ; classification ; genetics

This study was aimed to investigate the relationship between endostatin and vascular cell adhesion molecule-1 (VCAM-1) expressions on bone marrow stromal cells (BMSC) in mice after bone marrow transplantation (BMT) and effect of ligustrazine on their expressions. The mice were randomly divided into 3 groups: normal group (without treatment), saline group (control of BMT) and ligustrazine group (BMT + ligustrazine). BMT mouse models were established. The normal group was not treated, the saline group was given normal saline (0.2 ml/mouse, twice a day) through gastric tube, while the ligustrazine group was given ligustrazine (0.2 ml/mouse, twice a day) also through gastric tube. On day 7, 14, 21 and 28 after BMT, mice were killed by euthanasia. The expression levels of endostatin and VCAM-1 in bone marrow stromal cells were detected by immunohistochemistry and RT-PCR analysis respectively. The results showed that the endostatin protein mainly expressed in nuclei of BMSCs, the VCAM-1 protein mainly expressed in plasma of BMSCs. On day 7, 14, 21 after BMT the expression levels of endostatin mRNA and protein in ligustrazine and saline groups were significantly lower than that in normal group (P < 0.01 or P < 0.05), while their expression levels in ligustrazine group were lower than that in saline group. On day 28 the expression levels in saline group returned to normal, while the expression levels in ligustrazine group not were normalized. On day 7, 14, 21 after BMT the expression levels of VCAM-1 mRNA and protein in ligustrazine and saline groups were significantly lower than that in normal group (P < 0.01 or P < 0.05), but their expression levels in ligustrazine group were significantly lighter than that in saline group (P < 0.01 or P < 0.05). On day 28 the VCAM-1 expression level in ligustrazine group returned to normal, while its expression level in saline group not were normalized. The difference between these two groups was significant (P < 0.01). Correlation analysis revealed that there was a negative correlation between endostatin and VCAM-1 expression in saline group, there was a positive correlation between endostatin and VCAM-1 expression in ligustrazine group. It is concluded that the endostatin can influence hematopoiesis in bone marrow by affecting VCAM-1 expression on BMSC and hindering connection between stromal cells and hematopoietic cells as well as extracellular stroma and hematopoietic cells, while ligustrazine can enhance the adhesion molecule expression on stromal cell surface of bone marrow in BMT-mice, accelerate the homing and proliferation of HSPC in bone marrow after BMT, meanwhile can promote the repair of bone marrow microenvironment, accelerate hematopoietic reconstitution of bone marrow after BMT through feedback regulation of endostatin expression of BMSC in BMT-mice.
Animals ; Bone Marrow Cells ; cytology ; metabolism ; Bone Marrow Transplantation ; Endostatins ; biosynthesis ; genetics ; Female ; Male ; Mice ; Mice, Inbred BALB C ; Pyrazines ; pharmacology ; RNA, Messenger ; biosynthesis ; genetics ; Random Allocation ; Stromal Cells ; metabolism ; Vascular Cell Adhesion Molecule-1 ; biosynthesis ; genetics

To investigate the positive rate of anti-SARS antibody in children and adults without SARS, 197 paediatric patients under 14 years old from inpatient and outpatient department of our hospital, 156 healthy children pupils from primary school, 453 adult patients over 18 years old from inpatient and outpatient department of our hospital and other 502 healthy adult blood donors were selected. Anti-SARS antibodies were determined by anti-SARS specific antibody detection kit and ELISA method. The results showed that both the positive rates of IgG antibody in paediatric patients and healthy children were about 2% (4/197 and 3/156), while the positive rates in adult patients and healthy adults were about 0.2% (1/453 and 1/502). The difference between the positive rates of children and adults was significant (chi(2) = 11.61, P < 0.001). IgM antibody was negative in all the samples. It is concluded that the anti-SARS IgG antibody positive rate in children was obvious higher than that in adults. This may be the cause why the cases with SARS in children is much less than in adults.
Adult ; Age Factors ; Aged ; Antibodies, Viral ; blood ; Child ; Child, Preschool ; Female ; Humans ; Immunoglobulin G ; blood ; Male ; Middle Aged ; SARS Virus ; immunology

OBJECTIVESTo investigate the therapeutic effects and mechanism of octreotide on experimental hepatic fibrosis in rats.

METHODSHepatofibrotic rats models were established with carbon tetrachloride. All the experimental rats were divided into four groups: normal control group, pre-and post-treatment model group, and octreotide-treated group in which the rats were injected subcutaneously with octreotide at the dose of 50ng/100g, twice daily, for thirty days. Serum levels of hyaluronic acid (HA), laminin (LN) and pro-collagen type III peptide (PCIII) were detected by radioimmunoassay. Hepatic fibrosis scoring grade was assessed through Van-Gieson staining and observed under light microscope. Protein expression levels of alpha-smooth muscle actin (alpha-SMA) and transforming growth factor beta1 (TGFbeta1) were determined with immunohistochemical staining method. Messenger RNA (mRNA) levels of collagen type I and PCIII were detected by reverse transcription polymerase chain reaction.

RESULTSSerum levels of HA (ng/L), LN (microg/L) and PCIII (ng/L) in pre- and post-treatment model groups were higher than those in normal control group (121.8+/-9.5 and 110.3+/-13.4 vs. 33.1+/-3.7, 85.7+/-12.1 and 78.2+/-7.9 vs. 37.1+/-6.3, 35.9+/-3.5 and 33.7+/-2.6 vs. 15.6+/-2.8, respectively, t > or = 9.41, P<0.05), and there was no significant difference between the two model groups. Concentrations of HA (55.8ng/L+/-7.2ng/L), LN (43.1microg/L+/-3.4microg/L) and PCIII (27.8ng/L+/-3.4ng/L) decreased significantly in octreotide-treated group, compared with those in model groups (t >or=2.76, P<0.05). With histological analysis, fibrotic scoring grade in octreotide-treated group was obviously ameliorated, compared with that in model groups (chi2 > or = 3.97, P<0.05). Imaging analysis revealed that alpha-SMA and TGFbeta1 immunohistological staining areas were markedly shrinked in octreotide-treated group (t > or = 2.47, P < 0.05). In two model groups, PCIII and type I mRNA levels significantly up-regulated as compared with those in normal group (t > or = 9.27, P<0.001), and they were inhibited by octreotide markedly (t > or = 2.47, P<0.05).

CONCLUSIONSOctreotide can inhibit hepatic stellate cells transforming into myofibroblasts, down-regulate TGFbeta1, collagen type I and PCIII transcriptions, so that it has therapeutic effects on experimental hepatic fibrosis.

Actins ; analysis ; Animals ; Carbon Tetrachloride ; toxicity ; Collagen Type I ; genetics ; Collagen Type III ; genetics ; Hyaluronic Acid ; blood ; Laminin ; blood ; Liver ; pathology ; Liver Cirrhosis, Experimental ; drug therapy ; metabolism ; pathology ; Male ; Octreotide ; therapeutic use ; RNA, Messenger ; analysis ; Rats ; Rats, Sprague-Dawley ; Transforming Growth Factor beta ; analysis ; Transforming Growth Factor beta1

OBJECTIVETo detect potential mutations of OTC gene in a male infant affected with ornithine transcarbamylase deficiency.

METHODSGenomic DNA were isolated from peripheral blood samples of family members and 100 healthy individuals. Potential mutations of the 10 exons of OTC gene were screened with PCR and Sanger sequencing.

RESULTSA homozygous missense mutation c.917G>C in exon 9, which results in p.R306T, was identified in the infant. Sequencing of the mother and two female members of the family indicated a heterozygous status for the same mutation. The same mutation was not found in other members of the family and 100 healthy controls.

CONCLUSIONA missense mutation c.917G>C in the OTC gene is responsible for the pathogenesis of the disease. Identification of the mutation can facilitate prenatal diagnosis and genetic counseling for the family.

Computational Biology ; Female ; Humans ; Male ; Mutation ; Ornithine Carbamoyltransferase ; genetics ; Ornithine Carbamoyltransferase Deficiency Disease ; diagnosis ; genetics ; Sequence Analysis, DNA