NDV strain ZJ1 strain , a highly virulent NDV strain, has been prevalent among the waterfowls in China mainland in the past years. Multi-basic amino acid sequence distribute in the protease cleavage site of F protein of this strain. Recombinant expressing plasmid pCI-FT, was generated by converting multi-basic amino acid sequence of 112, 115, 117 of the protease cleavage site of F_ 0 protein, to the non-basic amino acid sequence characteristic of avirulent NDV strain. The result from co-expression of mutant or parental F protein with homologous HN protein in COS-1 cells revealed that both mutant and parental F protein had fusion activity. The result from co-expression of mutant or parental F protein with homologous HN protein in CEF cells showed that the cleavage activity of mutant F protein was significantly reduced. The study built a foundation for mutagenesis of amino acid sequence of the protease cleavage site of F_ 0 protein at the full-length cDNA clone level, study on factors contributing to virulence and construction of candidate vaccine strain, and so on.

OBJECTIVETo observe the impacts of different frequencies of electroacupuncture (EA) on post-operative cognitive function and the change in serum S-100beta protein under the compound anesthesia of acupuncture and drugs.

METHODSOne hundred and twenty-four patients of abdominal operation at selective time were randomized into a routine drug anesthesia group (group A, 24 cases), a meridian point 2 Hz group (group B, 26 cases), a me ridian point 2 Hz/100 Hz group (group C, 25 cases), a meridian point 100 Hz group (group D, 24 cases) and a transcutaneous acupoint electric stimulation 2 Hz/100 Hz group (group E, 25 cases). In group A, the endotrachea-lgeneral anesthesia was applied. In the rest groups, the acupuncture anesthesia was induced for 30 min before the endotracheal general anesthesia, at Baihui (GV 20), Yintang (GV 29) and Neiguan (PC 6), with G6805-2 electric acupuncture apparatus used. In group B, the continuous wave and 2Hz in frequency were selected. In group C, the disperse-dense wave and 2 Hz/100 Hz in frequency were selected. In group D, the continuous wave and 100 Hz in frequency were selected. In group E, the disperse-dense wave and 2 Hz/100 Hz in frequency were selected, and the electrode pads were stick on the acupoints and connected with the electric stimulation till the end of operation. Mini-mental state examination (MMSE) was adopted to evaluate and record the changes in cognitive function 1 day before operation and on the 3rd day after operation. The conditions of post-operative cognitive dysfunction (POCD) in the patients and the changes in serum S-100beta protein were monitored before and at the end of operation.

RESULTSThe incidence rate of POCD on the 3rd day after operation was 41.7% (10/24) in group A. The incidence rates of POCD were 26.9% (7/26), 16.0% (4/25), 33.3% (8/24) and 16.0% (4/25) in group B, C, D and E separately. Compared with group A, the incidence rate of PCOD in group B, C, D and E were reduced (all P<0.05), the incidence rate in group C and E were lower than that in groups B and D (all P<0.05). At the end of operation, the level of serumS-100beta protein was (0.186 +/- 0.027) microg/L in group A, the levels were (0.165 +/- 0. 028) microg/L, (0.166 +/- 0.027) microg/L, (0.163 +/- 0.025) microg/L and (0.164 +/- 0.025) microg/L in group B, C, D and E separately. The levels of serum S-100beta protein in group B, C, D and E were lower than that in group A separately (all P<0.05).

CONCLUSIONThe general anesthesia assisted with EA at different frequencies reduces the incidence of cognitive dysfunctionand, decreases the level of serum S-100beta protein after intestinal cancer resection. The effects of the meridian point electric stimulation at 2 Hz/100 Hz and the transcutaneous electric stimulation at 2 Hz/100 Hz are the best. Hence, these two approaches of anesthesia deserve to be recommended practically.

Abdomen ; surgery ; Acupuncture Analgesia ; Aged ; Aged, 80 and over ; Anesthesia, General ; Cognition ; drug effects ; Electroacupuncture ; Female ; Humans ; Male ; Postoperative Period

OBJECTIVETo determine Ligustilide in volatile oil from Ligustrcum chuanxiong with RP-HPLC.

METHODODS2 column (4.6 mm x 200 mm, 5 microm) was used and nitrendipine was used as internal standard. The mobil phase consisted methanol, acetontrile and water (33:21:46). The ligustilide was at 275 nm.

RESULTThe linear range was 2.92-29.2 mg x L(-1) for ligustilide. The average recovery of ligustilide was 95.1% and RSD was 2.3%.

CONCLUSIONThis method is simple and can be used to determine ligustilide with satisfactory accuracy and reproducibility.

4-Butyrolactone ; analogs & derivatives ; analysis ; Chromatography, High Pressure Liquid ; methods ; Ligusticum ; chemistry ; Oils, Volatile ; chemistry ; isolation & purification ; Plants, Medicinal ; chemistry ; Rhizome ; chemistry

OBJECTIVETo explore the effect of acupuncture-anesthetic composite anesthesia (AACA) on the incidence of postoperative cognitive dysfunction (POCD) and changes of TNF-alpha, IL-1beta, and IL-6 in elderly patients.

METHODSTotally 83 patients undergoing surgical resection of gastrointestinal tumor were randomly assigned to the simple anesthesia group (A group, 41 cases) and the AACA group (B group, 42 cases). Patients in Group A received endotracheal general anesthesia. Those in Group B were induced by acupuncture anesthesia for 30 min by needling at Baihui (DU20), Neiguan (PC6), Zusanli (ST36). The electro-acupuncture (EA) apparatus was connected after arrival of qi, with the wave pattern of density 2/100 Hz. The stimulus intensity was set by patients' tolerance, with the peak current of 5 mA. Then the endotracheal general anesthesia was performed and the EA lasted till the end of the surgery. The cognitive function of all patients was assessed before operation and at day 3 after operation using mini-mental state examination (MMSE). POCD was confirmed if with one or more decreased stand- ard. The peripheral venous blood was collected before anesthesia induction (TO), immediately at the end of surgery (T1), 24 h after operation (T2), and 48 h after operation (T3), and serum concentrations of IL-1beta, IL-6, and TNF-alpha were correspondingly measured using ELISA.

RESULTSThe postoperative anesthesia awakening time was shorter in Group B than in Group A [(20.37 +/- 6.09) min vs (29.24 +/- 7.48) min, P < 0.05]. The remifentanil dose used during the operation was less in Group B than in Group A (P < 0.05). The incidence of POCD at day 3 was lower in Group B than in Group A [10/41 (23.8%) vs 15/42 (36.5%), P < 0.05]. The concentrations of IL-1beta, IL-6, and TNF-alpha at T1-T3 were higher than those at TO in the two groups (P < 0.05). The increment of TNF-alpha and IL-1beta was less in Group B than in Group A (P < 0.05). CONCLUSION AACA could reduce the incidence of POCD and inhibit postoperative release of TNF-alpha, IL-1beta, and IL-6 in elderly patients undergoing colorectal cancer resection.

Acupuncture Analgesia ; Aged ; Cognition Disorders ; etiology ; prevention & control ; Gastrointestinal Neoplasms ; surgery ; Humans ; Interleukin-1beta ; blood ; Interleukin-6 ; blood ; Postoperative Complications ; prevention & control ; Tumor Necrosis Factor-alpha ; blood

Eight fragments were amplified and cloned into pCR2.1 vector with the designed primers.The fragments,amplified with primer Ⅰ to Ⅶ,were subcloned into transcription vector to construct the plasmid pNDVZJI which contained the full-length cDNA of NDV ZJI strain.The eukaryotic expression vector pCI-L was constructed by subcloning the fragments,amplified with the primer Ⅴ,Ⅵ and Ⅷ,into the expression vector pCI-neo.The full-length cDNA clone,pNDVZJI,with three helper plasmids,pCI-NP、pCI-P and pCI-L,were cotranfected into BSR-T7/5 cell expressing T7 RNA polymerase.After inoculation of transfected cell culture into embryonated chicken eggs from specific pathogen free(SPF)flock,The NDV of ZJI strain was rescued successfully,which laid a good foundation for the further related research.

Objective : To investigate the willingness to receive HIV testing in primary health service institutions (PHSIs) among young students in Wuhan City, so as to provide the evidence for improving the detection of HIV testing among young students. Methods: Fifteen PHSIs were sampled using a stratified random sampling method in 14 districts of Wuhan City, and school students at ages of 15 to 24 years were sampled from each district using a convenience sampling method. Participants' demographics, awareness of AIDS-related knowledge, HIV testing and willingness to receive HIV testing were collected using questionnaires, and factors affecting the willingness to receive HIV testing in PHSIs were identified among school students using a multivariable logistic regression model. Results : A total of 301 questionnaires were allocated, and 299 valid questionnaires were recovered, with an effective recovery rate of 99.34%. The respondents included 143 men (47.83%) and 156 women (52.17%), and had a mean age of (19.36±2.40) years; there were 223 respondents with an educational level of diploma and above (74.58%). The awareness of AIDS-related knowledge was 71.57% among the respondents, and 144 respondents had received AIDS-related health education in PHSIs (48.16%). There were 34 respondents that had received HIV testing (11.37%) and 203 respondents that were willing to receive HIV testing in PHSIs (67.89%). The respondents that were unwilling to receive HIV testing in PHSIs were mainly attributed to considering to be unlikely to get HIV infections (82.29%). Multivariable logistic regression analysis showed that school students who knew AIDS-related knowledge (OR=2.797, 95%CI: 1.583-4.941), knew free HIV counseling and testing services in PHSIs (OR=2.070, 95%CI: 1.123-3.814), and had received AIDS-related health education in PHSIs (OR=2.814, 95%CI: 1.573-5.032) were more willing to receive HIV testing in PHSIs. Conclusions There were 67.89% of school students that were willing to receive HIV testing in PHSIs in Wuhan City, and the willingness to receive HIV testing was correlated with the awareness of risk of HIV infections, and awareness and experience of AIDS control services in PHSIs.

Objective To observe whether the expression of interferon-regulatory factor genes are re- lated to systemic lupus erythematosus (SLE).Methods The clinical data of 45 SLE patients and 37 normal controls were collected.Total RNA of peripheral blood was extracted and transcripted into cDNA.Sybr green dye based real-time quantitative PCR method was used to compare the expression (indicated as-??Ct value) of IRFI,IRF4,IRF8 in patients with SLE and those in the controls.Results The levels of IRF1,IRF4 and IRF8 mRNA were-3.90?0.19,-8.04?0.25 and 3.60?0.15 respectively in normal controls.In SLE patients, IRF4 mRNA expression was -8.82?0.18,higher than that in normal (P=0.011).But IRF8 mRNA expression was 3.09?0.13,lower than that in normal (P=0.012).Conclusion Abnormal IRF mRNA expression is found in the peripheral blood of SLE patients.IRFs may play roles in the pathogenesis of SLE by affecting the differen- tiation of Th cells.

Based on the complete genome sequence of Newcastle disease virus (NDV) ZJI strain,seven pairs of primers were designed to amplify a cDNA fragment for constructing the plasmid pNDV/ZJI,which contained the full-length cDNA of the NDV ZJI strain.The pNDV/ZJI,with three helper plasmids,pCIneoNP,pCIneoP and pCIneoL,were then cotransfected into BSR-T7/5 cells expressing T7 RNA polymerase.After inoculation of the transfected cell culture supernatant into embryonated chicken eggs from specific-pathogen-free (SPF) flock,an infectious NDV ZJI strain was successfully rescued.Green fluorescent protein (GFP) gene was amplified and inserted into the NDV full-length cDNA to generate a GFP-tagged recombinant plasmid pNDV/ZJIGFP.After cotransfection of the resultant plasmid and the three support plasmids into BSR-T7/5 cells,the recombinant NDV,NDV/ZJIGFP,was rescued.Specific green fluorescence was observed in BSR-T7/5 and chicken embryo fibroblast (CEF) cells 48h post-infection,indicating that the GFP gene was expressed at a relatively high level.NDV/ZJIGFP was inoculated into 10-day-old SPF chickens by oculonasal route.Four days post-infection,strong green fluorescence could be detected in the kidneys and tracheae,indicating that the recombinant GFP-tagged NDV could be a very useful tool for analysis of NDV dissemination and pathogenesis.

AIM:To explore the effect of curcumin(Cur)and curcuminoids(Y20 and 6B)on the expression of secretory leukocyte protease inhibitor(SLPI), tumor necrosis factor-α(TNF-α)and interleukin-1β(IL-1β)induced by Streptococcus pneumoniae(SP)and the possible mechanism.METHODS:BEAS-2B cells incubated with SP were set up as an inflammation model of pneumonia.The mRNA levels of SLPI at 1 h,3 h,6 h and 9 h,and the mRNA expression of TNF-αand IL-1βat 3 h,6 h and 9 h in control group,SP infection group,Cur treatment group,Y20 treatment group and 6B treatment group were measured by qPCR.The protein levels of TNF-αand IL-1βin the culture supernatant were measured by ELISA.The protein levels of Toll-like receptor 2(TLR2)and phosphorylated nuclear factor-κB(p-NF-κB) p65 at 3 h,6 h and 9 h were determined by Western blot.RESULTS:The mRNA level of SLPI was increased in Cur, Y20 and 6B treatment groups compared with SP group(P<0.05).The protein levels of TLR2 and p-NF-κB p65 were sig-nificantly increased after SP stimulation.After treatment with Cur,Y20 and 6B,the protein levels of TLR2 and p-NF-κB p65 were significantly decreased(P<0.05).The levels of TNF-αand IL-1βwere significantly increased after SP stimula-tion.Cur,Y20 and 6B significantly decreased the levels of TNF-αand IL-1βin the supernatant(P<0.05).CONCLU-SION: Cur, Y20 and 6B increase SLPI expression, reduce the expression of inflammatory cytokines TNF-αand IL-1β. The possible mechanism might be associated with inhibiting TLR 2 expression and down-regulating the transcriptional activity of NF-κB.

OBJECTIVETo investigate the expressions of telomerase reverse transcriptase (TRT) and vascular endothelial growth factor (VEGF) and their correlation in prostate cancer (PCa).

METHODSTRT and VEGF expressions were assayed in 30 cases of PCa and 30 cases of benign prostatic hyperplasia (BPH) by means of immunohistochemistry (SP) combined with computer assisted image analysis.

RESULTSThe expression of TRT was detected in 19 of the 30 cases of PCa and 5 of 30 cases of BPH, and that of VEGF in 23 of the 30 PCa and 14 of the 30 BPH patients. TRT and VEGF expressions were significantly higher in cancer tissues than in BPH (P < 0.05). A significant correlation was observed between TRT and VEGF expressions (r = 0.8333, P < 0.05).

CONCLUSIONThe expression of TRT or VEGF might be a malignant phenotype in PCa. The expression of TRT is significantly correlated with that of VEGF, but the mechanisms are yet to be further studied.

Animals ; Humans ; Immunohistochemistry ; Male ; Prostatic Hyperplasia ; metabolism ; pathology ; Prostatic Neoplasms ; metabolism ; pathology ; Rabbits ; Telomerase ; biosynthesis ; Vascular Endothelial Growth Factor A ; biosynthesis