0.05). The positive rates of MMP-2 and MMP-9 were significantly higher in the cases with lymphnode metastasis [84.72% (61/72) and 77.78% (56/72)] than those without lymphnode metastasis [50% (4/8) and 37.50% (3/8), P

To carry out pathologic diagnoses and whole-genome sequence analyses of the Jaagsiekte sheep retrovirus (JSRV) in Xinjiang, China, we first observed sheep suspected to have the JSRV. Then, the extracted virus suspension was observed by transmission electron microscopy (TEM). Total RNAs from lungs of JSRV-infected sheep were extracted and reverse-transcribed using a cDNA synthesis kit. Six pairs of primers were designed according to the exogenous reference virus strain (AF105220). Reverse transcription-polymerase chain reaction was carried out from JSRV-infected tissue, and the whole genome of the JSRV sequenced. Our results showed: flow of nasal fluid ("wheelbarrow test"); different sizes of adenoma lesions in the lungs; papillary hyperplasia of alveolar epithelial cells; alveolar cavity filled with macrophages; dissolute nuclei in central lesions. TEM revealed JSRV particles with a diameter of 88 nm to 125. 4 nm. The full-length of the viral genome sequence was 7456 bp. BLAST analyses showed nucleotide homology of 96% and 95% compared with that of the representative strain from the USA (AF105220) and UK (AF357971). Nucleotide homology was 89.8% and 89.9% compared with the endogenous Jaagsiekte sheep retrovirus, Inner Mongolia strain (DQ838493) and USA strain (EF680300). The specific pathogenic amino-acid sequence "YXXM" was found in the TM district, similar to the exogenous JSRV: this gene has been reported to be oncogenic. This is the first report of the complete genomic sequence of the exogenous JSRV from Xinjiang, and could lay the foundation for study of the biological characteristics and pathogenic mechanisms of the pulmonary adenomatosis virus in sheep.
Amino Acid Sequence ; Animals ; China ; Genome, Viral ; Jaagsiekte sheep retrovirus ; classification ; genetics ; isolation & purification ; pathogenicity ; Lung ; pathology ; virology ; Molecular Sequence Data ; Phylogeny ; Pulmonary Adenomatosis, Ovine ; pathology ; virology ; Sheep ; Viral Proteins ; chemistry ; genetics ; Virulence

Objective:To explore the effect of early mother to child separation on neonatal nervous system and its related mechanism.Methods:Randomly selected during January 2015 to September 2015,was born in 120 cases of newborn mice as the research object,the newborn mice was born after the implemented of mother-to-child separation as observation group (60),not the implemented of separation of mother and baby after birth as the control group (60),in view of the two groups of newborn mice of the nervous system,the change of the nerve cells were compared and researched.Results:The apoptotic rate of neurons in the neonatal mice was significantly higher than that in the control group (P ＜0.05).The neurons of caspase-3 protein expression was significantly higher than the control group (P ＜0.05).The expression of Caveo-1 protein in the glial cells of 14 days and 21 days was observed in the neonatal mice.The expression of Caveo-1 protein in the glial cells of the control group For comparison,(P ＜0.05),which were statistically significant.Conclusion:The early implementation of matemal and infant birth separation of neonatal nervous system will produce great influence,and influence the expression of the nervous system in the development of newborn,which affect the behavior of the neonatal adult dysplasia.

Objective To improve the 2,3,5-triphenyl tetrazolium chloride (TTC) solution method for measuring the viability of Cistanche deserticola seeds and investigate the change in viability during storage at 5 ℃. Methods The effect of the testa,TTC concentration,sodium hypochlorite concentration (NaClO),and staining time were studied,and seed viability during storage at 5 ℃ was measured with the improved method. Results Seeds were kept for 48 h in 0.5% TTC solution at 40 ℃,and then for 2 h in 0.2% NaClO solution;Seed viability was measured under a stereomicroscope. Storing seeds of C. deserticola for 1 to 2 years at 5 ℃ had no significant effects on their viability. However,the percentage of seeds with high viability was increased with the extension of the storage time at 5 ℃. Conclusion A convenient and rapid method for measuring the viability of C. deserticola seeds is developed. Storing C. deserticola seeds at 5 ℃ could improve their viability

Objective To investigate the effect of hyperthermia on human hepatocellular carcinoma cell line SMMC-7721 after down-regulating the expression of mTOR,and its possible mechanisms.Methods An antisense mTOR (mammalian target of rapamycin) gene eukaryotic expression vector was transfected into SMMC-7721 cells.The expression of mTOR mRNA and protein were detected using RT-PCR and Western blotting,respectively.Hyperthermia was applied after the transfection,and the vitality of cell proliferation was evaluated using CCK-8 assays and the clone formation rate was determined by colony-forming assays.The migration of SMMC-7721 cells was measured using scratch assays.Apoptosis and the cell cycle were analyzed by flow cytometry.Results The expression of mTOR mRNA and protein were significantly decreased after transfection,indicating that the antisense vector could down-regulate the mTOR gene effectively.The proliferation,clone formation and migration of SMMC-7721 cells all were decreased markedly by hyperthermia after transfection.Flow cytometry showed that the rate of apoptosis was significantly increased.The number of cells in the S phase was increased and the cell cycle was induced to arrest at the S phase.Conclusions Down-regulating the expression of mTOR can increase the thermosensitivity of SMMC-7721 cells.The mechanism involves increased apoptosis and S phase arrest.

Objective　To perform the pathogenic and genomics analyses on isolates of Streptococcus suis (Ss) from three human infections in Shenzhen, aiming to provide a basis for the prevention and control of Ss outbreaks. Methods　The suspected bacterial strains from three blood plate cultures of three critically ill patients in three hospitals were subjected to biochemical identification, matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS), and real-time fluorescent PCR identification, resulting in the identification of three strains positive for Streptococcus suis serotype 2(SS2). Pure positive cultures were taken for an antimicrobial susceptibility test and extracted nucleic acids for whole-genome sequencing and analysis. The whole-genome sequencing and analysis included species identification, antibiotic resistance genes alignment, multilocus sequence typing (MLST), virulence genes alignment, and coregene-based phylogenetic tree analysis. Results　The blood agar isolates from three patients were all identified as Ss, the VITEK 2 identified them as SS2, and MALDI-TOF-MS identified them as Ss. Real-time PCR results for the universal gene gdh and serotype 2 cps2 gene of Ss were both positive. The antimicrobial susceptibility test results showed that all three strains were resistant to erythromycin and clindamycin, with variable sensitivity to tetracycline. Whole-genome sequencing results showed that all three strains were identified as Ss, including one ST7 strain and two ST1 strains. The virulence gene prediction results based on the VFDB database showed that all three strains were positive for mrp, sly, and cps, indicating high virulence gene characteristics. The analysis of the phylogenetic tree based on coregene showed that the three strains were in different evolutionary branches, with two ST1 strains having a closer evolutionary distance. Conclusions　The pathogens responsible for these three critically ill patients were SS2, and all three strains were resistant to erythromycin and clindamycin. Genetically, they all carried virulence genes that are found in highly virulent strains, while showed differences in MLST typing and phylogenetic tree analysis, indicating the presence of different genotypes of high pathogenicity SS2 in Shenzhen area and had caused sporadic cases, which requires high attention.

Aluminum salts are the most popular adjuvants applied in human vaccines currently. However,they can′t achieve satisfying results in the development of novel vaccines because of the cellular immune responses induced by them are weak and their adjuvant activities for some novel vaccines are poor, especially in vaccination against peptide antigens with small molecular weight. cGAMP (cyclic guanosine monophosphate-adenosine monophosphate) has recently been known as a mammalian second messenger, which plays an important role in the innate immune signaling pathway and is capable of boosting the immuno-genicity of vaccines,activating antigen-presenting cells and enhancing specific T cell responses. cGAMP is expected to become a new generation of vaccine adjuvants against infectious diseases and cancer. In this re-view,we summarize the application and current situation of vaccine adjuvants, describe the discovery of cGAMP and its mechanism as a vaccine adjuvant,and focus on the advances in using cGAMP in the fields of vaccination against infectious diseases, intradermal immunization and tumor immunotherapy. Finally, it is also pointed out that cGAMP,as a novel vaccine adjuvant,will have a broad prospect of application in areas such as anti-tumor,anti-virus,anti-inflammatory and vaccines.

ObjectiveTo investigate the clinical characteristics and prognostic factors in intrahepatic cholangiocarcinoma (ICC) patients with positive alpha-fetoprotein (AFP).MethodsClinical data of 57 patients with ICC in the Third Afifliated Hospital of Sun Yat-sen University from September 2004 to December 2013 were analyzed retrospectively. The informed consents of all patients were obtained and the ethical committee approval was received. There were 34 males and 23 females with an average age of (52±13) years old. According to serum AFP test result after admission in hospital, the patients were divided into AFP positive group (n=16) and AFP negative group (n=41). The difference of clinicopathological factors between two groups such as age, gender, hepatitis B surface antigen (HBsAg), liver cirrhosis, biliary lesions, preoperative carbohydrate antigen 19-9 (CA19-9), surgical procedure, metastastic lymph nodes, histological differentiation degree were compared byt-test , Chi-square test or Fisher's exact probability test. The patients were followed up and the survival of two groups was recorded. Prognostic factors analysis was conducted by Log-rank test and Cox proportional hazards regression model.ResultsMale patients accounted for 94% (15/16) in AFP positive group, which was signiifcantly higher than that in AFP negative group [46%(19/41)] (χ2=10.747,P<0.05). Patients complicated with liver cirrhosis accounted for 88% (14/16) , which was significantly higher than that in AFP negative group [37% (15/41)] (χ2=11.937,P<0.05). No patients were complicated with biliary lesions in AFP positive group but in AFP negative group, 12 cases were complicated with biliary lesions, where signiifcant difference was observed between two groups (P<0.05). Surgical procedure was the independent risk factor for patients in AFP positive group. Patients undergoing radical resection had better prognosis (RR=26.813,P<0.05).ConclusionsICC patients with positive AFP are mostly complicated with liver cirrhosis but a low incidence of biliary lesions. Surgical procedure is the independent risk factor for the prognosis. Radical resection is an effective way to improve the prognosis of ICC patients with positive AFP.

ObjectiveTo investigate the application value of Tacrolimus sustained-release capsules in patients after liver transplantation (LT).MethodsClinical data of 48 patients after LT treated in the Third Afifliated Hospital of Sun Yat-sen University between January 2011 and December 2013 were retrospectively studied. The informed consents of all patients were obtained and the local ethical committee approval had been received. Among the 48 patients, 43 were males and 5 were females with an average age of (45±12) years old. According to whether Tacrolimus sustained-release capsules were given after LT, the patients were divided into sustained-release group (18 patients) and control group (30 patients). The patients in both groups received combined immunosuppressive therapy of Tacrolimus + Methylprednisolone in the early stage after operation. Patients in sustained-release group were given Tacrolimus sustained-release capsules of equivalent dose 20 d after LT. The changes of plasma valley concentration of Tacrolimus and the incidence of rejection and adverse reactions of the two groups were compared. The comparison of plasma valley concentration of Tacrolimus was conducted usingt test, and the comparison of incidence of rejection and adverse reactions was conducted using Chi-square test or Fisher's exact test.ResultsThe plasma valley concentration of sustained-release group decreased gradually and stabilized after 30 d, while that of control group lfuctuated widely. On 60 d, the plasma valley concentration of sustained-release group was (7±2) μg/L, which was signiifcantly lower than (10±1) μg/L of control group (t=-15.61,P<0.05). The incidence of rejection of sustained-release group was 0 (0/18), which was signiifcantly lower than 7% (2/30) of control group (P<0.05). The incidence of pneumonia, gastrointestinal symptoms, hyperglycemia and sleep disorders of sustained-release group was respectively 33% (6/18), 28% (5/18), 6% (1/18) and 6% (1/18) , which was signiifcantly lower than 77% (23/30), 37% (11/30), 17% (5/30) and 23% (7/30) of control group (χ2=18.90, 31.53, 12.41, 45.19;P<0.05).ConclusionsThe application of Tacrolimus sustained-release capsules in patients after LT can maintain stable plasma valley concentration of Tacrolimus, have good anti-rejection effect and reduce the incidence of adverse reactions.

Objective To study the effects of Guizhi Shaoyao Zhimu Decoction on factors related to bone destruction and bone protection in rats with collagen-induced arthritis(CIA)based on osteopro-tegerin(OPG)/receptor activator of NF-κB ligand(RANKL)/receptor activator of NF-κB(RANK)signaling pathway.Methods According to the body weight,60 female Wistar rats were randomly di-vided into the following six groups:the normal group,the model group,the Triperygium wilfordii mul-tiglucoside group(0.01 g/kg),the Guizhi Shaoyao Zhimu Decoction low-dose group(8.6 g/kg),the Guizhi Shaoyao Zhimu Decoction medium-dose group(17.2 g/kg),and the Guizhi Shaoyao Zhimu Decoction high-dose group(34.4 g/kg)(n=10 rats per group).The rats in all groups except for the normal group were given 100 μg bovine type Ⅱ collagen on the 1st and 8th days to establish the CIA model,and was injected into the left foot sole and tail root of the rats.After the successful modeling,the rats were treated by gavage for 4 weeks.The general state,body weight,and arthritis index(AI)score of rats were recorded,and the contents of RANKL and OPG in rat serum were determined by enzyme-linked immunosorbent assay.The mRNA and protein expressions of RANKL,RANK,and OPG in the ankle joint were determined through real-time PCR and Western blotting,respectively.Results Com-pared with the normal group,the general state of the model group was poor,the toe swelling was obvious,the AI score was increased,the serum RANKL content was increased,the serum OPG content was de-creased,and the mRNA and protein expressions of RANKL and RANK in the ankle joint were increased(P＜0.05).Compared with the model group,the degree of toe swelling and the AI score of rats in the Guizhi Shaoyao Zhimu Decoction low-,medium-,and high-dose groups were decreased,the serum RANKL content was decreased,the serum OPG content was increased,the mRNA and protein expres-sions of RANKL and RANK in the ankle joint were decreased,the mRNA and protein expressions of OPG were increased,and the RANKL/OPG ratio of the ankle joint was decreased(P＜0.05).Conclusion Guizhi Shaoyao Zhimu Decoction can improve the destruction of joint bone in CIA rats,and its mecha-nism of action may be related to reducing RANKL level,reducing RANKL/OPG ratio,and regulating bone balance.