OBJECTIVEThe SSR information in the transcriptome of Erigeron breviscapus was analyzed in this study, in order to further develop new functional genes SSR markers laid a solid foundation.

METHODSSR loci were searched in all of 52,060 unigenes by using est_timmer. Perl program and SSR primers were designed by Primer3. Furthermore, 36 pairs of primers were randomly selected for the polymorphism analysis on 13 Erigeron breviscapus plants collected from different places.

RESULTA total of 3639 SSRs were found in the transcriptome of Erigeron breviscapus, distributed in 3260 unigenes with the distribution frequency of 6.99%. Di-nucleotide repeat was the main type, account for as much as 34.41% of all SSRs, followed by mono-nucleotide (31.41%) and tri-nucleotide repeat motif (28.08%). The di-nucleotide repeat motifs of AT/AT and AC/GT were the predominant repeat types (28.71%). The tri-nucleotide repeat motifs of AAT/AT was the predominant repeat types (7.94%). For validation the availability of those SSR primers, we randomly selected 36 pairs of primers for PCR amplification. Among them, 34 pair primers (94.44%) produced clear and reproductive bands, 19 pair primers showed polymorphism (52.78%), and 13 Erigeron breviscapus plants were divided into 2 groups.

CONCLUSIONThere are numerous SSRs in Erigeron breviscapus transcriptome with high frequency and various types, this will provide abundant candidate molecular markers for genetic diversity study and genetic map in this plant.

China ; DNA Primers ; genetics ; Erigeron ; classification ; genetics ; Genetic Variation ; Microsatellite Repeats ; Phylogeny ; Polymorphism, Genetic ; Transcriptome

The complete genomic sequence of Duck hepatitis virus 1 (DHV-1) ZJ-V isolate was sequenced and determined to be 7 691 nucleotides (nt) in length with a 5'-terminal un-translated region (UTR) of 626 nt and a 3'-terminal UTR of 315 nt (not including the poly(A) tail). One large open reading frame (ORF) was found within the genome (nt 627 to 7 373) coding for a polypeptide of 2 249amino acids. Our data also showed that the poly (A) tail of DHV-1 has at least 22 A's. Sequence comparison revealed significant homology (from 91.9% to 95.7%) between the protein sequences of the virus in the Picornaviridae family, its genome showed some unique characteristics. DHV-1 contains 3copies of the 2A gene and only 1 copy of the 3B gene, and its 3'-NCR is longer than those of other picornaviruses. Phylogenetic analysis to do sequence homology based on the VP1 protein sequences showed that the ZJ-V isolate shares high sequence homology with the reported DHV-1 isolates (from 92.9% to 99.2%), indicating that DHV-1 is genetically stable.

OBJECTIVETo design DNA microarray and investigate the molecular anti-tumor mechanism of herbs of traditional Chinese medicine.

METHODcDNA microarrays consisting of 56 probes representing 24 human cell cycle genes were constructed, Four anti-hepatocarcinoma herbs including Radix Linderae, Hebra Artemisiae Annuae, Radix Amebiae, Radix Astragli, were chosen. Effects of herbs on SMMC-7721 cell cycle were observed by flow cytometry assay. Effects of herbs on cell cycle gene expression in SMMC-7721 cells were analyzed by comparing hybridization of Dig-Labeled cDNAs from herb-treated cells and cDNAs from untreated cells.

RESULTExpressions of cell cycle geneswere changed in different degrees after herbs treated. Some genes were down-regulated and some genes were up-regulated. The changes in gene expression agreed with the results of flow cytometry assay.

CONCLUSIONThe results suggest that these herbs may have effects on cell cycle and DNA damage checkpoint genes which may be the mechanism of the herbs, and DNA microarray can be used to investigate the biological function of extracts of traditional Chinese medicine.

Antineoplastic Agents, Phytogenic ; isolation & purification ; pharmacology ; Artemisia ; chemistry ; Astragalus membranaceus ; chemistry ; Carcinoma, Hepatocellular ; metabolism ; pathology ; Cell Line, Tumor ; Cyclin-Dependent Kinase 4 ; Cyclin-Dependent Kinase Inhibitor p16 ; genetics ; metabolism ; Cyclin-Dependent Kinases ; genetics ; metabolism ; Drugs, Chinese Herbal ; isolation & purification ; pharmacology ; Gene Amplification ; Gene Expression Profiling ; Genes, cdc ; drug effects ; Humans ; Lindera ; chemistry ; Lithospermum ; chemistry ; Liver Neoplasms ; metabolism ; pathology ; Oligonucleotide Array Sequence Analysis ; methods ; Plants, Medicinal ; chemistry ; Proliferating Cell Nuclear Antigen ; genetics ; metabolism ; Proto-Oncogene Proteins ; genetics ; metabolism ; cdc25 Phosphatases ; genetics ; metabolism

BACKGROUND: Arthroscopic reconstruction for treatment of avulsion fracture at the tibial insertion of the knee posterior cruciate ligament (PCL) can minimize surgical trauma to the largest degree. However, its implants have relatively poor stability; therefore, functional exercise cannot be performed in the early stages after surgery, which is inconducive to knee function recovery. It is extremely challenging to perform traditional open reduction with internal fixation to repair avulsion fractures at the tibial insertion of the knee PCL. Often, the crushed bones cannot be firmly fixed, leading to a poor repair effect. OBJECTIVE: To design a new inverted "L"-shaped incision in the popliteal fossa through which bone plates were inserted to fix the crushed bones and to reconstruct PCL tension, facilitating knee function recovery; to compare the therapeutic effects of this new fixation and reconstruction method, and arthroscopic reconstruction for treating avulsion fracture at the tibial insertion of the knee PCL. METHODS: A prospective, single-center, non-randomized controlled trial. One hundred and eighty patients (knees) with avulsion fracture at the tibial insertion of the knee PCL will be assigned to two groups based on treatment methods: arthroscopic reconstruction group (n = 90;fracture fixation and repair under the arthroscope) and new method group (n = 90; bone plates will be inserted through an "L"-shaped incision in the popliteal fossa to fix the crushed bones and reconstruct PCL tension). After surgery, these patients will be followed up for 6 weeks, 6 months, and 12 months. RESULTS AND CONCLUSION: The primary outcome measure is the excellent and good rate of knee function recovery at 12 months after surgery as evaluated by Lysholm Knee Scoring Scale score (Herein referred to as Lysholm score). The secondary outcome measures are the excellent and good rate of knee function recovery before surgery, 6 weeks and 6 months after surgery; Lysholm score before surgery, 6 weeks, 6 months and 12 months after surgery; Hospital for Special Surgery (HSS) knee score, Visual Analogue Scale (VAS) score, posterior drawer test negative rate, X-ray morphology of the knee before surgery, and 6 weeks, 6 months, and 12 months after surgery and; incidence of adverse events at 6 weeks, 6 months and 12 months after surgery. Results of a preliminary study involving 62 patients (knees) with avulsion fracture at the tibial insertion of the knee PCL showed that posterior drawer test negative rate and Lysholm score were significantly higher in the new method group compared to the arthroscopic reconstruction group (P < 0.05) at 3 months after surgery. This study will be performed to compare the therapeutic effects of bone plate insertion through an "L"-shaped incision made in the popliteal fossa to fix the crushed bones and to reconstruct PCL tension, and traditional fracture fixation and repair under the arthroscope to treat avulsion fracture at the tibial insertion of the knee PCL. We believe that the former method will be superior to the latter one because it can fix the avulsion fracture more firmly, facilitating knee function recovery. This study was approved by Medical Ethics Committee of Cangzhou Central Hospital of China (approval No. 2017-120-01). This study will be performed in strict accordance with the Declaration of Helsinki formulated by the World Medical Association. Participants provided signed informed consent prior to participation in the study. This study was designed in December 2017. Patient recruitment and data collection will begin in April 2018. Patient recruitment will end in June 2019. Data analysis will be performed in August 2020. The study will be completed in October 2020. Results will be disseminated through presentations at scientific meetings and/or by publication in a peer-reviewed journal. This trial was registered with the Chinese Clinical Trial Registry (registration number:ChiCTR1800015026). The version of this study protocol is (1.0).

Objective To investigate the clinical significance of a new method,axis-line-distance technique(ALDT),in scoliosis measurement.Methods Thirty cases with idiopathic scoliosis were measured on two separate occasions by six observers with the Cobb technique and the ALDT on PACS workstation.The interval time between two measurement occasions were three weeks.The data were analyzed statistically with the paired-sample t-test.Results(1)Concerning intraobserver variance in two measurement occasions,the minimum variance,the maximum variance and the average variance were 0, 24.00?,5.71??1.54?for Cobb technique and 0,12.00 mm,(1.95?0.58)mm for ALDT.There were significant measurement differences for four observers with Cobb method 39.00??10.69?versus 36.50?? 10.63?,31.73??10.96?versus 37.30??9.65?,32.03??7.49?versus 27.86??9.00?,29.77??8.87? versus 34.20??7.26?,all P0.05].(2)Concerning interobserver variances in six observers,the average measurement variance was 5.07??0.35?for Cobb method,and(2.32?0.26)mm for ALDT. There were significant measurement differences for every observer using with Cobb method(36.63??10.30? versus 33.27??10.10?,39.00?10.69?versus 31.73??10.96?,32.03??7.49~ versus 29.78?? 8.87?,36.63??10.30?versus 39.00??10.69?versus 32.03??7.48?,39.00??10.69?versus 32.03?? 7.49?,all P

OBJECTIVETo evaluate the effect of a novel blockade technique for gastric cancer on blood-borne metastasis of gastric cancer cells to portal vein.

METHODSTwenty-three cases of gastric cancer were divided into routine operation group (8 cases intraoperatively without blockade technique) and blockade group (15 cases with blockade technique). Blood samples from portal vein pre- and intraoperatively, as well as gastroepiploic vein limited within the blockade area were obtained to detect CK19 mRNA expression by using RT-PCR technique.

RESULTSBefore the dissection of gastric lesion, the overall positive rate of CK19 mRNA expression in portal vein blood is 34.7% (9/23), including 37.5% (3/8) in routine operation group and 33.3% (5/15) in blockade group. While the course of tumor resection, those positive rates were 87.5% (7/8) in routine operation group and 6.7% (1/15) in blockade group respectively (P < 0.05). CK19 mRNA expression in the right gastroepiploic venous blood limited within the blocking area was all positive in 15 cases of blockade group.

CONCLUSIONThis blockade technique can be used effectively to block the intraoperative spread of gastric cancer cells, thus prevent blood-borne metastasis due to operative manipulation.

Aged ; Biomarkers, Tumor ; blood ; genetics ; Female ; Gastrectomy ; Humans ; Keratins ; blood ; genetics ; Ligation ; Male ; Middle Aged ; Neoplasm Metastasis ; prevention & control ; Neoplastic Cells, Circulating ; pathology ; RNA, Messenger ; blood ; Reverse Transcriptase Polymerase Chain Reaction ; Stomach Neoplasms ; blood ; pathology ; surgery ; Vascular Surgical Procedures ; methods

OBJECTIVETo carry out epidemiological study on an outbreak caused by E. coli O157:H7 infection in Jiangsu province in 1999.

METHODSEpidemiological, microbiological and moleculebiological methods were used to find out the source, route of transmission and risk factors.

RESULTS95 severe O157:H7 infected patients with acute renal failure in 9 counties and districts of 2 municipalities were reported in Jiangsu province, 1999 while 83 of the patients died with a death rate of 87.37%. Most patients were seen in mid or late June. The ratio of male to female was 1 to 1.44 and 88.42% of the patients were over 50 years old. 38 patients occurred in 2000 with 34 deaths. Major factors contributing to the outbreak would include without drinking tap water, eating leftover food, poor sanitary status in kitchen, not washing hands before meal and after bowl movement. 2 strain of O157:H7 was isolated from severe patients and 3 from diarrhea cases. Carrier rate among animals was up to 9.62% and 99.41% of the strains carried toxic gene. Strains isolated from feces of patients and animals belonged to the same colonies.

CONCLUSIONThis outbreak was severe which caused by O157:H7 and was first seen in China, which was closely related to the high carrier rate of O157:H7 in animals and to the positive rate of high toxic gene of the strains. There were various routes of transmission and the main factors of infection would include poor personal health habits and poor sanitation of the household.

Acute Kidney Injury ; epidemiology ; etiology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Antibodies, Bacterial ; immunology ; Case-Control Studies ; Child ; Child, Preschool ; China ; epidemiology ; Diarrhea ; epidemiology ; microbiology ; Disease Outbreaks ; Escherichia coli Infections ; complications ; epidemiology ; Escherichia coli O157 ; isolation & purification ; Escherichia coli Proteins ; immunology ; Female ; Hemolysin Proteins ; immunology ; Humans ; Infant ; Male ; Middle Aged ; Seroepidemiologic Studies

OBJECTIVETo investigate the food water sources of adults in the four cities of China in summer.

METHODSA total of 64 adults aged 18 - 60 yrs from four cities (Beijing, Shanghai, Chengdu and Guangzhou)were selected using convenient sampling method. The food samples were collected by using duplicate portion method and the water content of food samples were determined by using the national standard. The information on amounts and types of daily drinking soup was recorded by subjects using a quantitative measurement.

RESULTSA total of 63 subjects had completed the investigation. The median of daily water from food of subjects was 1157 ml, while water from staple food was 480 ml, the median ratio of which accounting for 41.8% of water from food. Water from dishes was 427 ml, the median ratio of which accounting for 37.9% of water from food. Water from soups was 133 ml, the median ratio of which accounting for 13.8% of water from food. Water from snacks was 0 ml, the median ratio of which accounting for 6.4% of water from food. The medians of water from staple food (384, 388 and 435 ml, respectively) in Beijing, Shanghai and Guangzhou was significant lower than that of Chengdu subjects' (900 ml, χ(2) = 21.27, P = 0.000). But the median proportion of water from staple food of subjects in Beijing (47.7%) was significantly higher than that in Chengdu (43.2%), Shanghai (42.9%) and Guangzhou (33.9%) (χ(2) = 8.69, P = 0.034). The median of amount and proportion of water from soups of subjects in Guangzhou (267 ml, 24.4%) and Chengdu(278 ml, 15.7%) was significantly higher than that in Shanghai (133 ml, 9.0%) and Beijing (100 ml, 5.8%) (amount: χ(2) = 22.52, P = 0.000;proportion: χ(2) = 16.27, P = 0.001).

CONCLUSIONThe staple food and dishes are the main sources of daily food water.

Adolescent ; Adult ; China ; Drinking ; Female ; Food Analysis ; Humans ; Male ; Middle Aged ; Seasons ; Surveys and Questionnaires ; Urban Population ; Young Adult

OBJECTIVETo study the responses of different pancreatic cancer cells to stimulations by nerve growth factor (NGF) and explore the role of Trk-A in such responses.

METHODSFive pancreatic cancer cell lines (MIA-PaCa-2, PANC-1, SW-1990, AsPC-1, and BxPC-3) were exposed to different concentrations of NGF (0, 4, 20, 100, and 500 ng/ml). MTT and Matrigel invasion method were used to observe the changes in the cell proliferation and invasion ability. Trk-A expression in these cells was detected by PCR and Western blotting, and the relations of Trk-A expression to the cell proliferative and invasive abilities following NGF treatment were analyzed.

RESULTSNGF at 100 ng/ml most obviously stimulated the cell proliferation, and PANC-1 cells showed the highest while AsPC-1 cells showed the least sensitivity to 100 ng/ml NGF stimulation. Matrigel invasion test showed that NGF enhanced the invasiveness of PANC-1 and MIA-PaCa-2 cells but produced only limited effect on AsPC-1 cells; the effect of NGF was completely inhibited by the Trk-A inhibitor CEP701. The expression levels of Trk-A mRNA and protein were the highest in PANC-1 cells and the lowest in AsPC-1 cells.

CONCLUSIONNGF can enhance the proliferation and invasiveness of pancreatic cancer cells, and this effect is possibly mediated by Trk-A protein.

Cell Line, Tumor ; Cell Movement ; drug effects ; Cell Proliferation ; drug effects ; Humans ; Neoplasm Invasiveness ; Nerve Growth Factor ; pharmacology ; Pancreatic Neoplasms ; metabolism ; pathology ; Receptor, trkA ; genetics ; metabolism

OBJECTIVETo compare differences of angiogenesis among collagen- chitosan, collagen-sulfonated carboxymethyl chitosan porous scaffolds and acellular dermal matrix after these three different scaffolds with silicone membrane were transplanted on the wounds of full thickness burn, and the wound repair of different scaffolds with epidermis grafting on.

METHODSAngiogenesis in different dermal scaffolds, the wound surface and epidermis survival were observed in 1, 2, and 3 weeks after the three different scaffolds were respectively transplanted on wounds of full thickness burn with debridement in 6 Bama miniature pigs (total 18 pigs in 3 groups). At the same time, CD34 positive signals (neo-forming microvessels) were detected by immunohistochemical staining. The wounds without any scaffold transplantation were studied as the control.

RESULTSAngiogenesis had been fundamentally finished in 2 weeks after implantation of collagen- sulfonated carboxymethyl chitosan porous scaffold. And fundamental angiogenesis in collagen- chitosan porous scaffolds and acellular dermal matrix needed at least 3 weeks. Neo-forming micro-vessels perpendicular to wound beds with these three different scaffolds were more than those in the control wounds without scaffold. CD34 positive signals (neo-forming micro-vessels) were significantly higher in wounds at the second week than those in wounds at the first week. And those in wounds at the third week were significantly higher than those in wounds at the second week in all wounds with different scaffold transplantations and the control wounds. CD34 positive signals in the group of sulfonated carboxymethyl chitosan porous scaffold on the 1st, 2nd and 3rd week after the scaffold transplantation were significantly higher than those corresponding signals in the other three groups. Epidermis on the sulfonated carboxymethyl chitosan porous scaffold which had been transplanted on burn wound for 1 week could survive perfectly, however, epidermis on the collagen- chitosan porous scaffold or acellular dermal matrix could not survive until these two scaffolds had been transplanted on the burn wounds for at least 2 weeks.

CONCLUSIONSThese three different scaffolds could repair the full thickness skin defects caused by burn, and angiogenesis of sulfonated carboxymethyl chitosan porous scaffold is the best.

Animals ; Burns ; surgery ; Chitosan ; analogs & derivatives ; Collagen ; Disease Models, Animal ; Female ; Silicones ; Skin Transplantation ; Skin, Artificial ; Swine ; Swine, Miniature ; Tissue Scaffolds