Objective To investigate the evaluation for condition and prognosis of butyrylcholinesterase(BuChE)evaluation for acute organophosphorus pesticide poisoning (AOPP) .Methods 110 cases of AOPP patients were collected ,venous blood of all the patients in pre-hospital was sampled for determination of BuChE activity ,and treatment lasted for 60 d ,then BuChE characteristic curves and forecast indexs were analyzed .Results 24 cases died and 96 cases survived ,pre-hospital BuChE activity of the death group was (36 .7 ± 34 .2)% ,and the survival group was (67 .2 ± 15 .4)% ,there was significant difference between the two groups (P<0 .05) .That BuChE activity≤30% of normal value in ROC curve was the optimal cut-off point to determine the AOPP poi-soning .Among 27 cases of patients ,who BuChE activity ≤30% of normal value ,the death rate was 74 .07% (20/27) ,among 93 cases of patients ,who BuChE activity>30% of normal value ,the death rate was 4 .30% (8/93) ,there was significant difference between the two groups(P<0 .01) .When the optimal cut-off point of BuChE activity was≤30% of normal value in pre-hospital patients with critical AOPP ,the specific degree of its death prediction was 0 .954 ,sensitivity was 0 .761 ,positive predictive value was 0 .825 ,accuracy was 0 .912 ,Youden index was 0 .716 .Conclusion BuChE activity is a rapid and effective predicted value for the AOPP ,it is worthy of clinical application .

Tuberculosis (TB), an infectious disease caused by Mycobacterium tuberculosis (Mtb) infection,remains a major global concern.The activation of anti-TB immune responses is mediated by several complicated mechanisms, among which the local immune microenvironment at the disease site plays a crucial role.In this review, we discuss the roles of anti-TB immunity, Mtb immune escape and the elements at the local site of infection.

ObjectiveTo compare three types of acute Mycobacterium tuberculosis infection mouse models established through different infection routes and to set up the theoretical basis for further developing,selecting and applying these animal model in the tuberculosis-related research.MethodsStandard strain of Tubercle bacillus H37Rv was diluted to 1 × 106 colony forming unit (cfu)/mL.The mice were infected with the bacteria through different routes including intravenous injection,intranasal administration and inhalation of bacteria aerosol.Six weeks after the infection,the mice were euthaniz ed and necropsied. The lung tissues were collected and gross changes were observed.The colony counting was performed and the lung tissues were assessed by HE staining,acid fast staining.The e xpression level of tumor necrosis factor (TNF)-α per unit area in lung tissue was detected by immunohistochemistry. The data were analyzed by t test. Results The amounts of Mycobacterium tuberculosis in lung tissues of mice in inhalation group,intranasal administration group and intravenous injection group were (6.290±0.028),(6.150±0.021) and (6.120±0.008) lg cfu/mL,respectively; while no Mycobacterium tuberculosis was detected in control group. The difference between infection group and control group was statistically significant (t =3.762,P＜0.01),while there were no significant differences among infection groups with different infection routes (P＞0.05).According to the results of gross observations and histological assessment,the pathological changes were observed and red tubercle bacillus was detected by acid-fast staining in the lung tissues of all the mice in infection group.The results of immunohistochemistry showed that the expression levels of TNF-α per unit area were as follows:intravenous injection group (0.049 × 106 )＜intranasal administration group(0.759×106) ＜ inhalationgroup(1.042×106), whichwere statistically different (t =2.504,P＜ 0.05).ConclusionInhalation of bacteria aerosol may be the most efficient method to establish tuberculosis infection mouse model compared to intravenous injection and intranasal administration.

Objective To construct mycobacterial membrane-anchored expression vector and to analyze expression level and sub-cellualr localization of exogenous target protein. Methods Based on the mycobacterial intracellular expression vector pMFA42 which contained a strong promoter of pfurAma mutant, the signal sequence of Mycobacterium tuberculosis(Mtb) 19×103 lipoprotein (19SS) was synthesized and was then cloned into the downstream of pfurAma mutant to generate the mycobacterial membrane-anchored expression vector pMFA42M. The coding gene of enhanced green fluorescent protein(EGFP) was amplified by PCR, and then sub-cloned into these two vectors described above to construct recombinant EGFP fused and membrane-anchored strains, respectively. The coding genes of Mtb immuno-dominant antigens Ag85A and its chimera Ag856A2 were then sub-cloned intothe membrane-anchored construct pMFA42MG to produce recombinant Mtb antigen EGFP fused-expression strains. After that, expression levels and sub-cellualr localization of exogenous target protein were further analyzed by Western blot and flow cytometry sorting(FCS), and the fluorescence intensities of recombinant EGFP- expressed strains were observed in vitro directly and after transfection of murine macrophage cell line RAW264.7. Results The novel mycobacterial membrane-anchored expression vector was constructed successfully by introduction of signal sequence of Mtb 19×103 lipoprotein. Using of EGFP as model antigen, exogenous target protein was demonstrated to be expressed with high level and could be anchored into cell membrane of recombinant mycobaterial strains. Conclusion A novel mycobacterial membrane-anchored expression vector was constructed successfully to research recombinant BCG and functions of mycobacterial membrane proteins, and the constructed EGFP-expressed recombinant strains could also be used to research cytophagy in cell model and mycobacterial colony and translocation in animal immunization as model indicator bacteria.

Objective To investigate the influence of Qiliqiangxin capsule on the exercise tolerance in patients with chronic heart failure. Methods 86 patients with chronic heart failure were selected and recruited into a control group and an observation group randomly. The patients in the control group were given angiotensin-converting enzyme inhibitor, diuretic,carvedilol or metoprolol and dixina. The patients in the observation group were given Qiliqiangxin capsule on the basis of the control group. The change of exercise tolerance and left ventricular ejection fraction in all patients were detected after 4 weeks.Results The distance of walking in 6 minutes increased for all patients after 4 weeks' treatment(P＜0.05), while the observation group increased more significantly than the control group(P＜0.05). The left ventricular ejection fraction increased for all patients after therapy 4 weeks (P＜0.05), while the observation group increased more obviously than the control group (P＜0.05). The total excellent rate and effective rate in the observation group were both higher than the control group (P＜0.05). Conclusion Qiliqiangxin capsule can increase the clinic curative effect by increasing the exercise tolerance and improving the heart function of patients with chronic heart failure.

Objective To prepare the chitosan-potD nanoparticles and to evaluate its protective efficacy against pneumococcal nasopharyngeal colonization. Methods potD gene was amplificated from pneumococcal genome and was inserted into pVAX1 expression vectors to construct pVAX1-potD recombinant plasmid which was then transfected into 293T cell using LipofectAMINE 2000 to analyze transient potD gene expression in vitro by RT-PCR and Western blot. Chitosan-potD nanoparticles were freshly prepared by coacervation methods at each time and the characterizations of the nanoparticles were then evaluated. BALB/c mice were immunized with chitosan-potD, naked potD DNA or pVAX1 for 4 times at two-week intervals. Anti-PotD IgG, IgG1 and IgG2a levels in serum and IgA levels in nasal washes, bronchoalveolar lavage fluids (BALF) and middle ear lavages(MEL) were detected by indirect enzyme-linked immunosorbent assay (ELISA). IL-17A, IL-4 and IFN-γ levels in splenocytes were determined by double sandwich ELISA. Mice were intrannsally challenged with Streptococcus pneumoniae ATCC6303, and Pneumococci were recovered from the nasopharyngeal niche at the fifth day after challenge. Results potD gene was successfully amplificated by PCR and the sequence was confimed to be consistent with that in the Genbank. The pVAX1-potD recombinant plasmid was successfully constructed and was expressed in eukaryocytes in vitro. The mean size and zeta potential of chitosan-potD nanoparticles was 430 nm and + 20.5 mv, respectively. Chitosan-potD nanoparticles were not digested by DNase Ⅰ , while naked potD DNA was completely digested. The levels of antibodies inculding IgG, IgG1, IgG2a, IgA and cytokines including IL-17A, IL-4 and IFN-γ were significantly higher in mice immunized with chitosan-potD nanoparticles than mice with naked potD or pVAX1 ( P <0.05) only. More importantly, much less Pneumococci were recovered from mice immunized with chitosan-potD nanoparticles than the other groups(P <0.05). Conclusion Chitosan-potD nanoparticles significantly enhanced the immunogenicity and protection efficacy of DNA vaccines by intranasal immunization and could be used as a potential mucosal vaccine to prevent pneumococcal infection.

BACKGROUND: The training task for aircrew of army airmen is very heavy, the evaluation of flight personnel on psychological diathesis is performed to early discover and deal with corresponding questions.OBJECTIVE: To compare the difference of mental health between aircrew and ground crew of army airmen and provide reference data for implementing corresponding norms.DESIGN: Chester sampling investigation and paired design were applied.SETTING: the 71687 Force of Chinese PLA, the 71521 Force of Chinese PLA, Department of Epidemiology, Fourth Military Medical University of Chinese PLAPARTICIPANTS: By chester sampling, 122 aircrew and 122 ground crew of one army airmen in one stationed training troop were randomized in February 2001. The pairs were made at ratio of 1:1 according to age difference less than 3 years and same sex and nationality. The investigated persons were all male, of Han and aged varied from 23 to 52 years.METHODS: Cornell health questionnaire was adopted respectively in investigation of aircrew and ground crew of army airmen. The investigated persons finished and handed in questionnaire independently on the spot in manner of secretion and centralized filling-up. Cornell health questionnaire is composed of 18 sections, including 195 questions, involving 4 aspects,named somatic symptoms, medial and family history, general health and habit and mental symptoms. The total score of Cornell health questionnaire ≥ 30 indicates somatic and mental disturbance and the score of mental section ≥ 10 indicates mental disturbance, both of which are taken as the criteria in comparison of psychological health in receptors of two groups.MAIN OUTCOME MEASURES: Every factor score and total score in somatic and mental sections in Cornell health questionnaire for aircrew and ground crew.RESULTS: The tested results of each 122 cases in either aircrew group or ground crew group entered result analysis. Evaluated results of somatic and mental symptoms in receptors of two groups: the total score was low in Cornell health questionnaire (0-14 scores), of which, the total score in aircrew group was higher than that in ground crew group (81.5% and 59.8%,P ＜ 0.05); the score in mental section was low (0-9 scores), of which, that in aircrew group was higher remarkably than that in ground crew group (99.2% and 91.8%, P ＜ 0.01); the score for the receptors with somatic and mental disturbance (total score≥30, score in mental section ≥ 10) in aircrew group was lower than those in ground crew group (4.1% and 12.3%, P ＜ 0.05); the score for the receptors with mental disturbance (score in mental section≥ 10) in aircrew group was lower remarkably than ground crew group (0.8% and 8.2%, P ＜ 0.01). For somatic section in aircrew group, except the factors of muscle, skeleton and skin, their scores were higher than those in ground crew group (P ＞ 0.05), the scores of the rest factors were all lower than those in ground crew group, of which, the difference was significant in evaluation of factors of eye, ear, nervous system, urinary reproductive system and fatigue in comparison of two groups (P ＜ 0.05). The score of every factor in mental section of aircrew group was lower than those in ground crew group. Except tension factor, in comparison of the rest factors in two groups, the significant difference was present (P ＜ 0.05).CONCLUSION: Overall psychological health state in Cornell questionnaire of aircrew in army airmen was superior to ground crew. Aircrew still presents a part of psychological health questions, mainly in somatic section and inadaptability was the most positive response in mental section, suggesting that aircrew probably presents inadaptable phenomena.

Objective To construct DNA vaccine expressing Mycobacterium tuberculosis(Mtb) immunodominant antigen Ag85A and analyze its anti-tuberculosis T cell responses in BCG primed-mice after DNA vaccination boosting.Methods The coding gene of Ag85A mature fragment was amplified by PCR with H37Rv genomic DNA as template,and then cloned into the eukaryotic expression vector pVAX1 to construct Ag85A DNA vaccine.After purification,Ag85A DNA vaccine was injected intramuscularly twice in BCG primed-mice with BCG vaccination and DNA vaccination alone as control.Eight weeks post-vaccination,spleen lymphocytes were separated and were then used to analyze Mtb antigen specific effector T cell response and polyfuntional IFN-γ/TNF-α/IL-2 secreting CD4+ T cell frequencies and intensities,and CD8+T cell responses by IFN-γ ELISPOT assay and intracellular staining,respectively.Results Compared to BCG vaccinated-and DNA vaccinated-mice,Ag85A DNA boosting not only enhanced significantly BCG primed-mice IFN-γ+TNF-α+IL-2+,IFN-γ+ IL-2+,TNF-α+IL-2+ and IL-2+ CD4+ T cell frequencies and IL-2 secretion,but also improved significantly IFN-γ-secreting and IL-2-secreting CD8+ T cell frequencies.Condusion Ag85A DNA vaccine was constructed successfully and was demonstrated to enhance significantly BCG primed-mice Mtb antigen specific CD4+ and CD8+ T cell responses when boosting,which is beneficial to improve BCG immunogenicity and its waning immune protection against Mtb.

Objective To investigate the expression of CD8 and CD28 lymphocytes in peripheral blood of patients with breast cancer before and after operation and explore its clinical significance.Method Flow cytometry was applied to test the expression of CD8,CD8+CD28+and D8+CD28-in the peripheral blood of both 40 patients with breast cancer and 25 healthy controls. Results The expression levels of CD8 in patients with breast cancer were significantly higher than those in healthy controls [(30.4±3.3)%VS. 30.4±3.3)%,P<0.05] . The expression levels of CD8+CD28+in patients with breast cancer were significantly lower than those in healthy controls [(10.4 ± 1.7)%VS. (14.3±2.6)%,P<0.05] . The expression levels of CD8+CD28-in patients with breast cancer were significantly higher than those in healthy controls [(25.1 ±2.8)% VS. (19.6 ±2.3)%, P<0.05] . After operation, the expression levels of CD8, CD8+CD28+and CD8+CD28-[(31.9 ±4.3)%, (15.4 ±2.7)%and (20.4±2.8)%, respectively] in peripheral blood of patients with breast cancer had no significant change compared with the healthy controls. The expression levels of CD8, CD8+CD28+and CD8+CD28-in peripheral blood of patients with breast cancer had significant difference between before and after operation. Conclusions After surgical operation, the immune defence and immune surveillance of cytotoxic T lymphocytes can be enhanced, immune suppression and tumor escape can be reduced. The assays of cell phenotypes CD8 and CD28 might be useful for evaluating the immunal state and prognosis of patient s with breast cancer.

Objective To evaluate the clinical effect of the simple syringomyelia patients with syringopleural shunt (SPS) and syringosubarachnoid shunt (SSS).Methods Twenty-eight patients with syringomyelia were selected as our subjects.Of which,18 patients were operated with SPS and 10 cases were with SSS.The therapy effect was compared between groups.Results All patients were checked with MRI 3 months after the operations and showed that syrinx cavity was significantly narrow of all patients.At early stage (48 h) after surgery 9 cases in SPS and 4 cases in SSS were showed the decrease syrinx cavity.Among patients with SPS,15 cases (83.3％) had the symptoms and signs improved,1 case (5.5％) withno changes,1 case (5.5％) with worse effect,and 1 cases(5.5％) occurred the tethered spinal cord.Meanwhile,among,patients with SSS,8 cases(80.0％) had the symptoms and signs improved,1 case(10.0％) with no changes,and 1 case (10.0％) with worse effect.There were 4 patients with pneumothorax in SPS group after operations,and the lung compression ratios were less than 5％.These cases were not taken any special treatment for the pneumothorax.All patients in two groups had not been infected and pleural effusions.No cases had taken the tube plugging.Conclusion The simple syringomyelia patients with the spine injury should take the cavity shunt.The SPS has some advantages because it can provide the partly negative pressure.But it should be certified by more cases and a long-time follow up.