Objective To investigate the effects of nitric oxide (NO) inhalation on the expression of TNF-?,IL-8 and CD11b mRNA in lung tissue during acute respiratory distress syndrome (ARDS) induced by intravenous injection of endotoxin in dogs.Methods Twelve pure bred beagle dogs of both sexs weighing 8-12.5 kg were randomly divided into 2 groups: NO group received mechanical ventilation with NO inhalation (n = 6) and control group received only mechanical ventilation ( n = 6) . Sepsis and ARDS were induced by intravenous injection of endotoxin as described in detail in our previous paper. Hemodynamics and pulmonary oxygenation were monitored and shunt fraction was calculated. At the end of experiment the animals were sacrificed and lung tissue was obtained aseptically and stored in the liquid nitrogen at - 180℃ . The total RNA was extracted. Reverse transcription-polymerase chain reaction (RT-PCR) was used to detect the expression of TNF-?,IL-8 and CD11b mRNA. The total RNA was reversely transcripted to cDNA. Then the cDNA was amplified by PCR. The product of PCR was scanned by gel-image analysis system.?-action was used as internal control. Semi-quantitative method was adopted for measurement of TNF-? ,IL-8 and CD11b mRNA expression. Results The expression of TNF-?, IL-8 and CD11b mRNA in lung tissue was significantly decreased in NO group compared with those in control group.Conclusion NO inhalation reduces expression of TNF-?, IL-8 and CDllb mRNA in lung tissue during ARDS induced by intravenous endotoxin.

Objective To investigate the changes in pharmacokinetics of propofol and propofol concentration in the brain induced by normovolemic hemodilution ( NHD) . Methods Thirteen healthy male mongrel dogs weighing 10-15 kg were randomly divided into 2 groups: control group ( n = 7) and NHD group ( n = 6). The animals were anesthetized with intravenous diazepam 0.5 mg?kg-1 and ketamine 5 mg?kg-1 . The femoral artery and vein were cannulated. Lactated Ringer's solution was infused at 5-7 ml ? kg-1 ? h-1 . 30 min after induction of anesthesia NHD was conducted by removing blood from femoral artery and replacing it with 6% hydroxyethyl starch (HES) until Hct was reduced to 25% . Propofol was then infused at 10 mg?kg-1 ?h-1 for 30 min after NHD. Arterial blood samples were taken immediately before and at 1, 2, 5 , 10, 15, 20, 30, 31, 32, 35, 40, 50, 70, 90, 120, 150, 180, 240 and 300 min after propofol infusion was started for determination of plasma propofol concentrations. One week later the same NHD was repeated. Propofol was administered by TCI via Graseby 3500 infusion pump incorporated with Stanpump TCI software. Target plasma propofol concentration was set at 5 ?g?ml-1 . Arterial blood and CSF samples were obtained at 5, 15, 30, 45 and 60 min after the start of propofol infusion for determination of plasma propofol concentration ( bound and free) , plasma free propofol concentration and CSF propofol concentration. At the end of 60 min, after the collection of blood and CSF samples, brain tissue was obtained from the cerebral cortex of right frontal lobe for determination of brain propofol content. ECG, BP, arterial blood gases and body temperature were monitored during experiment.Results The plasma propofol concentrations were significantly lower during and after propofol infusion at 10 mg? kg-1 ? h-1 in NHD group than in control group ( P

Acupuncture Therapy ; methods ; Adult ; Female ; Humans ; Jaw ; physiopathology ; Male ; Middle Aged ; Temporomandibular Joint Disorders ; physiopathology ; therapy ; Young Adult

Objective To observe the clinical efficacy of superficial needling in treating temporomandibular joint disorder. Method Sixty patients with temporomandibular joint disorder were randomized into a treatment group and a control group, 30 in each group. The treatment group was intervened by superficial needling, and the patients were asked to repeatedly open and close mouth after the treatment; the control group was intervened by regular acupuncture. The treatment was given once a day, 10 d as a treatment course. The clinical efficacies were compared after 2 treatment courses.Result After the first treatment course, the total effective rate was 89.7% in the treatment group versus 86.2% in the control group; after the second course, the total effective rate was 96.6% in the treatment group versus 93.1% in the control group. There was a significant difference in comparing the therapeutic efficacy between the two groups after the second course (P<0.05). The relapse rate was 5.0% in the treatment group versus 33.3% in the control group, and the difference was statistically significant (P<0.05).Conclusion Superficial needling is an effective method in treating temporomandibular joint disorder.

Objective To study the tumor targeting ability and application of farnesylthiosalicylic Acid (FTS) and heptamethine carbocyanine fluorescent dye-mediated near-infrared imagine in living animals, and confirm the inhibitory effect of this compound on growth of tumor cells.Methods Human breast cancer cell line MCF-7, glioma cell line U251 and prostate cancer cell line PC3 were cultured to logarithmic growth phase, and different concentrations of FTS and FTS-IR783 were added, respectively.We observed the inhibitory effect of those two compounds on the growth of tumor cells.Under fluorescence microscopy, specific accumulation of FTS-IR783 in these tumor cells was observed.The tumor cells (1×106) were transplanted subcutaneously into nude mice.These mice were subjected to intraperitoneal injection of FTS-IR783 (10 nmol/mouse) two weeks later.In the in vivo imaging, near infrared fluorescence signal and tumor volume were measured and their correlation was analyzed.Results Compared with FTS, FTS-IR783 significantly inhibited the growth of MCF-7, U251 and PC3 cells in vitro.FTS-IR783 was specifically uptaken by these three kinds of tumor cells, showing strong near infrared fluorescence in cell agglomerates.After subcutaneous injection of FTS-IR783, the correlation between fluorescence intensity and tumor volume was 0.987, 0.998 and 0.971, respectively.Conclusions The compound of FTS conjugated with near infrared fluorescent dye IR-783 can specifically recognize tumor cells, in both in vitro and in vivo imaging.At the same time, the compound can significantly inhibit the growth of tumor cells, and may be expected to become a new potential targeted drug.

Objective To investigate the immunological properties of Rv1009 domain. Methods BALB/c mice were immunized with Rv1009 domain three times at 2-week interval. ELISA was used to detect the antiRv1009 domain antibody titer in the sera of immunized mice sera. The spleen lymphocytes of the immunized mice were separated and the stimulation index (SI) was measured by MTT colorimetry. Levels of secreted IFN-γ, IL-10 and IL-12 upon specific antigen stimulation were detected by ELISA. The BALB/c mice immunized with Rv1009 domain were intravenously infected with MTB H37Rv. Four weeks after the final injection, the number of CFU in spleens was determined. Results The titer of the specific antibody in sera of the immunized BALB/c mice was 1:12 800. The SI of Rv1009 domain immunized group (2. 40±0. 18) was significantly higher than that of saline immunized group (0.90±0.21). The IFN-γ,IL-10 and IL-12 levels in culture supematant of spleen lymphecytes from the fusion proteins immunized mice was (1 432±30) ng/L, (503±11) ng/L and (311±11) ng/L respectively, significant different from that of saline immunized group[(256±20) ng/L, (76±6) ng/L and(56±8) ng/L,P<0.01]. Four weeks after the final injection,compared with normal saline immunized mice (6.64±0.13), dramatic reduction in MTB replication was observed in the spleen (4.86±0.14) from BALB/c mice immunized with fusion proteins following a subsequent MTB H37Rv challenge, but the protection efficacy of mice immunized with Rv1009 domain was not as good as that of BCG vaccination group (3.81±0.16). Conclusion Rv1009 domain can be used as a candidate for the new TB vaccine.

Optical molecular imaging is more and more widely used in the field of biomedical sciences due to its advantageous properties such as non-invasive, real-time and high resolution. As a kind of important optical molecular imaging probe,the near-infrared fluorescent(NIRF)dyes exhibit less tissue absorption and strong tissue penetration,and has been gradually applied to the early diagnosis of tumors. Researchers have developed a number of NIRF dyes with high potential for clinical application by conjugating tumor-targeting ligands,nano-modifications and multimodal NIRF imaging, and have significantly improved the specificity and sensitivity of these NIRF dyes in tumor diagnosis. In this paper we provide a review on the application of NIRF dyes in tumor diagnosis.

Objective To purify native and recombinant heparin-binding hemagglutinin(HBHA)protein,and investigate the activity of HBHA polyclonal antibody against aggregation of Bacillus CalmetteGuerin(BCG)induced by HBHA.Methods After growing BCG to the stationary phase in the 7H9 liquid medium,the native HBHA protein(nHBHA)was obtained by CL-6B column chromatography.At the same time,the HBHA gene fragment was cloned and expressed by transforming Escherichia coli BL-21.Then the polyclonal antibody against rHBHA was prepared by immunizing rabbit.Different comcentration of the HBHA protein was added to the BCG liquid medium,and the aggregation of the BCG was observed.Then,add the HBHA protein that incubated with anti-HBHA antibodies to the BCG culture medium and observe the aggregation of BCG.Results The purity of native HBHA was 99% and the concentration was 1.016 mg/ml.The expressed product contained 36% of total somtic protein.After purified,the purity of the recombinant HBHA protein was 97.1% and the concentration was 10.98 mg/ml.Both the rHBHA and nHBHA could induce the aggregation of BCG.When then concentration of nHBHA is 0.2μg/ml,BCG could be induced to aggregate,while the rHBHA concentration is 2μg/ml could induce the aggregation.Both aggregations could be suppressed by the polyclonal antibody against rHBHA.Conclusions The native and recombinant HBHA are successfully obtained.It is proved that the rHBHA could induce the aggregation of BCG similar as nHBHA,and polyclonal antibody against rHBHA could also suppress the activity of nHBHA.It suggested that rHBHA could be further used in clinical diagnosis and vaccination.

Objective To purify Micrococcus luteus Rpf and Rpf domain fusion protein, and to in-vestigate its effects on growth of Mycobacterium tuberculosis. Methods The recombinant plasmids pPro-EXHT-Rpf and pPro-EXHT-Rpf domain were expressed in E. Coli DHSa and then purified under denaturing condition via Ni-NTA purification system and confirmed by Western blot. The biochemical property of the M. Luteus Rpf and Rpf domain was analyzed by stimulating the resuscitation of M. Tuberculosis H37Ra which were in non-culturable' condition. Results The Rpf and Rpf domain products achieved 95% and 93% pure respectively, and the molecular weight was 30 x 103 and 12 x 103, the yield of purification was about 471 mg/L and 337 mg/L of the culture. The M. Luteus Rpf and Rpf domain from the E. Coli showed activity of stimulating the resuscitation of M. Luteus and M. Tuberculosis H37Ra in non-cuhurable' condition which could be inhibited by monoclonal antibodies of M. Luteus Rpf domain remarkably. Conclusion It was dem-onstrated that the purification of Rpf and Rpf domain have high biological activity for further functional, pharmacological and clinical investigations, and M. Luteus Rpf domain protein is fully active as M. Lateus full-length Rpf.

Objective To screen monoclonal antibodies (mAbs) for early diagnosis of invisive Aspergillus. Methods Monoclonal antibodies against different antigens of Aspergillus fumigatus were produced. The two pairs of combinations of monoclonal antibodies were selected accoring the distinct epitopes and double-antibody sandwich ELISA based on mAbs above were established. The sensitivity and specificity of the methods were analyzed by detecting culture supernatants of clinical isolates and environmental isolatesof Aspergillus. spp, Penicillium Marneffei, Candidas, and serum from animal models and patients. The epitopes recognized by mAbs were identified by immunobotting. Results A total of 32 hybridoma cell lines that stably produced MAbs were obtained. Two double- antibody sandwich ELISAs were established. One method was specific for 19 clinical isolates and environmental isolates of Aspergillus. spp, whereas the other one was specific for the clinical and environmental isolates of Aspergillus fumigatus without cross-reation with other Aspergillus. spp. For the same kind of medium of Aspergillus fumigatus, the sensitivity of the first method was 10 fold higher than the second method. Conclusions The specific mAbs for early diagnosis of invisive Aspergillus were obtained. Antigen recognized by the specific mAbs was mannoprotein with molecular weights of approximately 25 000-75 000. This antigen was potential early diagnostic marker for invasive Aspergillus.