Natural killer T (NKT) cells,a subset of lymphocytes that bridge innate and adaptive immune systems,involve in processes of infection immunity,tumor immunity,transplantation immunity and autoimmunity.A significant progress has been made in the mechanisms of origin,selection,differentiation and maturation of NKT cells.However,some viewpoints are still controversial,and need to be further intensively investigated.The potential therapeutic applications of functional NKT cells have been suggested in the prevention and the treatment of various diseases.

The present study was performed to explore the role of corticotropin releasing factor (CRF) in the nucleus basalis of Meynert (NBM) in spatial learning and memory of rats. The latency, distance and swimming path to find the platform were measured by Morris water maze after intra-NBM injections of 0, 0.01, 0.1 and 0.4 nmol of CRF. Intra-NBM injections of 0.1 or 0.4 nmol of CRF induced significant increase of the latency for spatial learning and memory, and there were no significant changes in the swimming speed in Morris water maze test. The results suggest that CRF plays an inhibitory role in spatial learning and memory consolidation in the NBM of rats.

The pGenesil-1-Beclin1 eukaryotic expression vectors were constructed to establish an SH-SY5Y cell line stably expressing shRNA-Beclin1. The shRNA was connected to pGenesil-1 to construct the recombinant plasmid pGenesil-1-Beclin1, which was transformed into JM109 E. coli. Positive clones were identified by digestion with restriction endonuclease and DNA sequencing. SH-SY5Y cells were cultured by the conventional method. The pGenesil-1-Beclin1 and pGenesil-1 plasmids were transfected into SH-SY5Ycells, and the cells were screened by G418 until the stable G418-resistant monoclonal cells were acquired. Beclin1 mRNA and Beclin1 protein were detected by RT-PCR and Western blot analysis respectively. The results of restriction endonuclease analysis and DNA sequencing confirmed the correct construction of the eukaryotic expression vector pGenesil-1-Beclin1. Two SH-SY5Y transfected cell lines were successfully selected. Compared with the control group, RT-PCR and Western blot showed that the expression of Beclin1 mRNA and protein were down regulated 71.28% ± 1.45% (P < 0.05)and 75.50% ± 2.63% (P < 0.05), respectively. The results indicated that the eukaryotic expression vector pGenesil-1-Beclin1 was successfully constructed and the SH-SYSY cell lines with inhibited Beclin1 expression were established. It provides a useful cell model for studying the biological function of Beclin1.
Apoptosis Regulatory Proteins ; genetics ; metabolism ; Beclin-1 ; Cell Line, Tumor ; Down-Regulation ; Escherichia coli ; Gene Silencing ; Humans ; Membrane Proteins ; genetics ; metabolism ; Neuroblastoma ; Plasmids ; RNA, Messenger ; RNA, Small Interfering ; Transfection

Objective To observe CT features of struma ovarii (SO) in comparison of pathologic findings. Methods CT features of 9 patients with pathologically confirmed SO were analyzed retrospectively and compared with pathological findings. Results All tumors were unilateral and had smooth margins, 3 were cystic and 6 were cystic-solid lesions, 5 were high attenuation lesions in the cyst portion of the mass on contrast CT. The cyst wall moderately (n=3) or markedly (n=5) enhanced after administration of contrast medium. Scattered, stripe-like or ring-like calcifications were found in 6 patients. On pathology, most of cystic portions were filled with high proteinaceous gelatinous fluid of eosinophiclic colloid, and the solid portion of tumors consisted of the thyroid tissue and stoma containing abundant blood vessels and fibrous tissue. Conclusion SO appears as a smooth marginated multicystic mass with calcification, high attenuation lesionin plain CT, and marked solid part enhancement on contrast CT.

Objective To analyze the phenotypic characteristics of antiviral-resistant influenza B viruses circulating in mainland China and to analyze the susceptibility of influenza B viruses to neuraminidase inhibitors ( NAIs) . Methods Antiviral-resistant phenotyping test was performed to analyze the NAI suscep-tibility of 1 386 influenza B viruses isolated in mainland China from April 2014 to March 2015, including the test of susceptibility to oseltamivir and zanamivir. Results All of the 94 B-Victoria lineage viruses were sensitive to oseltamivir and zanamivir. Of all 1 292 B-Yamagata lineage viruses tested, 1 virus showed re-duced sensitivity to oseltamivir with NA gene containing I221T amino acid mutation, 10 viruses showed re-duced sensitivity to zanamivir with 4 having D197N amino acid mutation in NA gene, 3 viruses showed re-duced sensitivity to both oseltamivir and zanamivir with NA gene possessing D197N amino acid mutation and 1 virus carrying the A245T amino acid mutation in NA gene showed reduced sensitivity to oseltamivir and highly reduced sensitivity to zanamivir. Conclusion The majority of influenza B viruses circulating in main-land China during 2014 to 2015 were sensitive to NAIs, which indicated that NAIs could be used continually for clinical treatment of patients with influenza. Sustained monitoring of antiviral susceptibility of influenza B viruses should be emphasized for timely detection of antiviral resistant viruses and more attention should be paid to the D197N mutations in NA gene of influenza B viruses.

Objective To establish an inactivated viral particle-based ELISA for the detection of antibodies against Middle East respiratory syndrome coronavirus (MERS-CoV) in serum samples collected from a MERS-CoV associated case. Methods Serum samples were collected from 10 newborns and 40 healthy adults. A viral particle-based ELISA was established by using the inactivated MERS-CoV virions as antigen. The levels of IgM and IgG antibodies in the serum samples were detected by the established ELISA and the cut-off values for positive detection were determined. Then the inactivated MERS-CoV virion-based ELISA was used to detect the antibodies against MERS-CoV in 5 serum samples collected from the first im-ported MERS case in China. Results The cut-off values of IgM and IgG antibodies in serum samples for ELISA were determined to be A450 readings of 0. 32 and 0. 42, respectively. The titers of IgM and IgG anti-bodies in serum samples collected at early admission to hospital from the first imported MERS case in China were both 1 ︰ 40. Seroconversion occurred 2 weeks after his admission to hospital with the titers of IgM and IgG reaching to 1 ︰ 320. Conclusion The inactivated MERS-CoV virion-based ELISA was established successfully and could be used for the detection of serum antibodies (IgG and IgM) in MERS associated cases.

Objective:To observe the effect of electroacupuncture (EA) pretreatment on adenine nucleotides in the myocardial tissues of the myocardial ischemia-reperfusion injury (MIRI) rats, and to explore the mechanism of EA pretreatment on myocardial prevention and protection in MIRI rats. Methods:Forty SPF male Sprague-Dawley (SD) rats were randomly divided into 5 groups: a blank group, a sham operation group, a model group, an EA at Neiguan (PC 6) group and an EA at Hegu (LI 4) group, with 8 rats in each group. Rats in the blank group only received binding to the rat plate, 30 min/time, once a day for 7 d; on the 7th day, rats in the sham operation group were subjected to threading for 40 min at the left anterior descending coronary artery without ligation, and then the rats were allowed to stand for 60 min before collection of the specimens; on the 7th day, rats in the model group were subjected to threading at the left anterior descending coronary artery with ligation, for 40 min before the blood flow was restored, and then the rats were allowed to stand for 60 min before collection of the specimens; on the 7th day of pretreatment with EA at Neiguan (PC 6) or Hegu (LI 4) for 30 min per day (once a day for 7 d), rats in the EA at Neiguan (PC 6) group and EA at Hegu (LI 4) group were subjected to modeling and sample collection same as in the model group. The left ventricular myocardium of the lower left anterior descending coronary artery was collected from rats in all 5 groups. Hematoxylin-eosin (HE) staining and transmission electron microscope (TEM) were used to observe the changes in myocardial pathological morphology. The change in the adenine nucleotide level of myocardial tissue was measured by high performance liquid chromatography (HPLC). Results:The HE staining and ultrastructure showed that the myocardial injury was severer in the model group compared with the sham operation group. Compared with the model group, the myocardial injury in the EA at Neiguan (PC 6) and the EA at Hegu (LI 4) groups was mild or hardly any. The adenine nucleotide levels in the sham operation group and the model group were all decreased compared with the blank group (allP<0.05); compared with the sham operation group, the adenine nucleotide level of the model group was also decreased, but the difference was not statistically significant (P>0.05); compared with the model group, the adenine nucleotide level in the EA at Neiguan (PC 6) group was increased (P<0.05), and the adenine nucleotide level in the EA at Hegu (LI 4) group was significantly increased (P<0.01). The adenine nucleotide level in the EA at Hegu (LI 4) group was higher than that in the EA at Neiguan (PC 6) group, but the difference was not statistically significant (P>0.05). Compared with the EA at Neiguan (PC 6) group, the levels of adenosine triphosphate (ATP), adenosine diphosphate (ADP) and adenosine monophosphate (AMP) in the EA at Hegu (LI 4) group were significantly increased (allP<0.01). Conclusion:Both EA at Neiguan (PC 6) and Hegu (LI 4) can alleviate the pathological damage to myocardium in MIRI rats, and increase the adenine nucleotide level in myocardial tissues, and thus protect MIRI rats. EA at Hegu (LI 4) has a better protective effect than Neiguan (PC 6).

Objective To analyze the willingness and influencing factors for usage of pre-exposure prophylaxis(PrEP) among male students who have sex with men (MSM) in universities of Guangxi. Meth-ods 295 MSM students in universities were recruited by Snow-balling methods. Questionnaires were self-administered to collect social demographic information,AIDS related risky sexual behaviors and willingness for usage of PrEP. Logistic regression was employed for univariate and multivariate analysis. Results Of the 295 MSM,58(19.7%)had ever heard PrEP,265(89.8%) said that they would like to use PrEP. Multivari-ate logistic regression analysis showed that those who had found partners through friends ( OR=11.419,95%CI:1.363~95.641), those who would advise his friend to use PrEP ( OR=87.946,95%CI:13.660-566. 222),those who concerned the convenience to gain the medicine ( OR=119.652,95%CI:3.765-3802.184) and those who said that they could take medicine every day ( OR=88.245,95%CI:10.237-760.696) were more likely to accept PrEP. The subjects whose partners would be angry if they stick to use condoms( OR=0.106,95%CI:0.019-0.606),and those who suspected the effectiveness of drugs( OR=0.010,95%CI:0. 001-0.112) were less likely to accept PrEP. Conclusion The acceptability of PrEP is high in university students'MSM. The safety of drugs and the convenience to gain the medicine are the main influencing factors for usage of PrEP.Using of PrEP should be strengthened to the MSM.

To analyze the antigenic and genetic characteristics of the influenza A (H3N2) virus in mainland China during the surveillance year of 2013-2014, the antigenic characteristics of H3N2 virus were analyzed using reference ferret anti-sera. The nucleotide sequences of the viruses were determined by Sanger dideoxy sequencing, phylogenetic trees were constructed with the neighbor-joining method, and the genetic characteristics of the viruses were determined in comparison to current vaccine strains. The results showed that most of the H3N2 viruses were antigenically closely related to the A/Victoria/361/2011 vaccine strain cell-propagated prototype virus (99.6%). Using the A/Texas/50/2012 egg isolate as the reference antigen, 15.1% of the viruses were found to be closely antigenically related to it, while 11.9% of strains were closely antigenically related to the egg-propagated epidemic strain, A/Shanghai-Changning/1507/2012. Phylogenetic analysis of HA genes indicated that the A(H3N2) viruses in this surveillance year were in the same clade, but no drug resistant mutation was identified in the NA genes. During the 2013-2014 influenza surveillance year, no significant genetic change was detected in either the HA or NA genes of the A(H3N2) viruses, while significant mutations were found in egg isolates resulting from their adaptation during propagation in eggs. The antigenic and genetic changes should be investigated in a timely manner to enable the selection of an appropriate vaccine strain in China.
Animals ; Antigenic Variation ; Base Sequence ; Chick Embryo ; China ; Genetic Variation ; Hemagglutinin Glycoproteins, Influenza Virus ; genetics ; immunology ; Humans ; Influenza A Virus, H3N2 Subtype ; genetics ; immunology ; isolation & purification ; Influenza, Human ; virology ; Molecular Sequence Data ; Mutation ; Phylogeny

ObjectiveTo investigate the development trend of drinking water type of endemic fluorosis in Qinghai province, and to provide the basis for the prevention and treatment of the disease. MethodsIn 2009, six monitoring counties were chosen by using simple random sampling methods, all diseased villages of the six monitoring counties were classified into light, moderate and severe disease types according to water fluorine content on the historical data, and 1 village was respectively chosen from each type. In monitoring villages with improved water, 3 tap water and one source water samples were collected, respectively. Five water samples were collected randomly in water unimproved monitoring villages according to water well locations of east, west, south, north and center. The fluorine content in water and urine was determined according to the Standard Testing Methods for Drinking Water (GB/T 5750-2006). Children aged 8 to 12 were examined for dental fluorosis by Dean method.Clinical osteofluorosis of all the resident over the age of 16 was examined, 2 village of these counties were randomly selected, and clinically diagnosed patients with skeletal fluorosis were examined again by X-ray using Diagnostic Criteria of Endemic Skeletal Fluorosis (WS 192-2008). Urine sample of 30 children aged 8 to 12 and of 20 adults over the age of 16 were randomly collected and urinary fluoride was determined by F-ion selective electrode method (WS/T 89-2006). ResultsImproving water projects had been implemented in 14 monitoring villages of the 18 villages in 6 counties, the rate of improved-water was 77.78％(14/18). Among the 14 projects, 5 improved-water projects ran normally, and 9 projects ran with intermittently water supply. Seventy-five water samples were tested, themean of water fluoride was 0.48 mg/L. The prevalence of dental fluorosis was 31.95％ (285/892), that of clinical skeletal fluorosis was 36.55％(1570/4295) and the X-ray detection rate of skeletal fluorosis was 25.64％ (20/78).Five hundred and seventy-one urine samples of children were determined, and geometric mean of urinary fluorine was 1.04 mg/L; 370 adult urine samples were determined, and geometric mean of urinary fluorine was 1.52 mg/L Conclusion Epidemic of drinking water type of endemic fluorosis is still serious in Qinghai province, and drinking water defluoride measures should be further strengthened and improved.