Objective To establish a DNA barcode method for cell species identification, analyze and verify the applicability of the method, so as to use it for cell species identification in biological products production.Methods The mitochondrial cytochrome C oxidase subunit 1(COI) gene was selected as the target gene, and two sets of primers(V primer and L primer)were chosen for specific amplification of this target gene. The amplification products were sequenced, and their sequences were aligned with the database to determine the species of cells being detected.Results The two sets of primers exhibited differential amplification abilities across various cell species. V primer and L primer successfully detected cells from mice,hamsters, guinea pigs, gophers/prairie dogs, cats, cows, chickens, and ducks. Additionally, V primer could also be utilized for detecting cells from monkeys, rats, dogs, mink, pigs, rabbits and humans. L primer could also be utilized for the detection of insect-derived cells. This method presented a detection sensitivity of 10 cells and could detect cross contamination at a hostto-contaminated cell ratio of 10∶1. Review verification demonstrated that four units successfully identified the species of eight unknown cells using this DNA barcode method.Conclusion The established DNA barcode method is effective for identifying cell species from 17 different types, providing a new technical approach to ensure the accurate use of cells in biological products production.

Clinical research is to provide guidelines for the clinical practice.First clinical re search should follow the ethics,and the researcher should have correct scientific attitude.Furthermore clinical research should be closely integrated with clinical practice and innovated with its feasibility taken into account.

Objective To investigate the ultrasonographic features of normal rectal walls.Methods Ten removed rectal specimens were scanned with high frequency (4-13 MHz) linear array probe to obtain ultrasound images of various layers and having each layer marked,and separated,followed by histological examination respectively.Results Ultrasound demonstrated seven layers of structure which were identified by alternative high and low echoes.From innermost layer towards the outer layers,they were divided as:high-echo acoustic interface,low-echo mucous layer,high-echo sub-mucous layer,low-echo circular muscle layer,high-echo fibrous connective tissue layer,low-echo outer longitudinal muscle layer and high-echo outer membrane layer.All these findings were justified by histological examination.Conclusions High-frequency ultrasound demonstrated 7 layers of echoes in normal rectal walls.This provides imaging basis for diagnosis and judge the invasion degree of rectal cancers.

OBJECTIVE:To observe the hypoglycemic effects of Huangsang oral solution.METHODS:Diabetes models of mice was induced with alloxan then the mice were randomly divided into the NS group,model control group,phenformin group,and high dose,middle dose and low dose Huangsang oral solution groups(30.0 g?kg-1,15.0 g?kg-1 and 8.0 g?kg-1).The corresponding drugs were given by ig bid for 10 days.Blood glucose was measured with glucose oxidas method.The effects of Huangsang oral solution on blood glucose concentration and glucose tolerance in diabetic mice as well as on blood glucose concentration in the normal mice were observed.RESULTS:The different dose of Huangsang oral solution could decrease the blood glucose level significantly and improve glucose tolerance in alloxan-induced diabetes mice,and the effect was significant compared with the model control group(P

Objective To evaluate Jiangxia umbilical therapy on the quality of life in patients with gastric carcinoma chemotherapy after surgery.Methods Patients with gastric carcinoma chemotherapy after surgery in the Third Affiliated Hospital of Sun Yat-Sen University from January 2016 to December 2016 were chosen as the research subjects.According to the time of chemotherapy,the patients were divided into two groups.Patients with gastric carcinoma chemotherapy in hospital from January 2016 to June 2016 were chosen as control group,while patients in hospital from July 2016 to December 2016 were chosen as observation group.Before chemotherapy,the control group was given tropisetron hydrochloride intravenous injection,while the observation group added Jiangxia umbilical on the basis of this.Nausea and vomiting in the two groups were observed after treatment.Quality of Life Questionnaire of Stomach 22 was applied to analyze and compare the patients' quality of life.Results The incidence rate of anti nausea in the observation group was 88.10％,which in the control group was 61.90％,the difference was statistically significant (x2 =9.571,P ＜ 0.05).The overall quality of life in observation group was significantly better than that in the control group [(20.34 ±:5.84) points vs.(14.32 ± 5.97) points,t =8.686,P ＜ 0.01].The scores in the observation group on the quality of life in the aspects of total score,anxiety scale,reflux scale,eating restricted scale,dry mouth,tastes changing had statistically significant differences compared with the indicators in the control group (all P ＜ 0.05).Conclusion Jiangxia umbilical therapy can not only relieve the chemotherapy-induced nausea and vomiting of patients with gastric carcinoma chemotherapy after surgery,but also can improve their quality of life.

0.05).But in the group EDN and CDN,Lp(a) was significantly higher as compared with the former two groups (P

Objective To construct the bait plasmid of pSos-single immunoglobin IL-1 receptor related protein (SIGIRR) in Cy-toTrap yeast two hybrid system ,and to test its self-activation .Methods The cDNA fragments of SIGIRR(480 -1 230 bp) were amplified from pReceiver-LV19-SIGIRR and ligated into the bait plasmid pSos to generate the plasmid pSos-SIGIRR .The pSos-SI-GIRR was identified by DNA sequencing and dual-site endonuclease digestion .Then the recombinant plasmid and control plasmid were introduced into the yeast cell cdc25H .The transformants were inoculated on plates of 25 ℃ /SD/Glucose(-UL) ,25 ℃/SD/Ga-lactose(-UL) ,37 ℃ /SD/Glucose(-UL) and 37 ℃ /SD/Galactose ,respectively and the proliferation ability of transformant was ob-served for 6 d .The Western blot was adopted to detect the expression of target protein .Results The pSos-SIGIRR vector was cor-rectly constructed and proved of no self-activation and toxic action .The Western blot showed that the target protein was expressed in a form of fusion protein of 170KD .Conclusion The bait plasmid containing SIGIRR cytoplasmic tail can be applied to the yeast two-hybrid system and lays the important foundation for seeking the interacting protein with SIGRR from the human lung cDNA li-brary in .

Objective To investigate the surgical treatment of infected femoral pseudoaneurysm caused by injection of heroin. Method Retrospective analysis was made on the clinical data of 14 cases of infected ruptured femoral pseudoaneurysm caused by injection of heroin. Results All 14 cases underwent operation. The ruptured sizes of arteries were 0.5 cm-2.0 cm . All cases were treated by resection of the aneurysm and replacement of iliac-femoral artery with artificial graft. Blood supply and function were good in all limbs after operations. Aneurysm cavities and incision wounds were treated with debridement, drainage and anti-infective treatment. All wounds were healed. 2 weeks and 3 months after operation, the grafts were patency demonstrated by Color Doppler examination. None of them was complicated with anastomosis leakage or thrombosis. Conclusion Thorough debridement of aneurysms and reconstruction of the artery are the effective approach in treating the infected ruptured femoral pseudoaneurysm caused by injecting heroin. Sufficient drainage, anti-infective and anti-coagulation therapy should be considered after the operation.

Abstract] Objective To establish an assay for the detection of porcine parvovirus ( PPV) and to verify its application for monitoring cells used for production.Methods A pair of primers and one probe were designed according to the conserved sequence encoding non-structural protein 1 (NS1).Based on the designed primers, a real-time fluorescent quantitative PCR assay for the detection of PPV was developed. Several parameters including the linearity, precision, minimum detection limit and anti-interference of the established assay were evaluated.A stock of PPV strains was prepared by infecting swine testicle ( ST) cells with PPV strains.An assay for the detection of PPV infection was developed by using ST cells as sensitive cells.A combined ST cell infection-PCR test was developed by combining the ST cell infection assay with the real-time fluorescent quantitative PCR assay.The sensitivity of ST cell infection-PCR test was analyzed.The cell samples used for production of biological products were detected by using the established assay.Results The real-time fluorescent quantitative PCR assay was specific for the detection of PPV without cross-reaction to other species of parvovirus virus, SV40 virus and other porcine viruses.The linear range of the assay was 1×109-1×104 copies/μl with a R2 value more than 0.98.The sensitivity of the real-time quantitative PCR assay was 1×104 copies/μl.Both of the intra-and inter-coefficient of variation (CV) were less than 5%in Ct values.The intra-and inter-CV in copies of detection were 5%-15% and 30%-40% respectively.The minimum detection limit of the real-time fluorescent quantitative PCR assay was 1CCID50/ml.The PPV strains were detected in cell samples with no interference.The sensitivity of ST cells infection-PCR test was 0.01CCID50/ml.All of the 22 cell samples were negative for PPV by using the real-time fluorescent quanti-tative PCR assay.Conclusion The real-time fluorescent quantitative PCR and the ST cell infection-PCR test for the detection of PPV in cells were established successfully.The application of the two assays was conducive to further enhance the safety of using cells for production and therapy.

Objective:To investigate the effects of nitric oxide donor,sodium nitroprusside(SNP)on the expression and production of TLR4 in U937 induced by LPS.Methods:U937 was induced to maturation by PMA and then stimulated by LPS.Then the cells were treated with SNP and divided into four groups:control group without stimulation of LPS,LPS group(10 ng/ml LPS+100 ng/ml rhLBP),low dose SNP group and high dose SNP group(50,500 ?mol/L SNP respectly).The mRNA and protein of TLR4 was determined by RT-PCR and Western blot.Results:The mRNA of TLR4 in SNP groups was lower than those in LPS group(0.308?0.050 and 0.138?0.0044 vs 0.342?0.098,P0.05).Conclusion:SNP might have a potential protective role in LPS induced inflammation such as sepsis and acute lung injury through inhibiting the mRNA and protein expression of TLR4.