Application of video laryngoscope combined with bronchial blocker and double-lumen tube in the standardized training of anesthesia for tracheal intubation / 中华医学教育探索杂志

Objective:To explore the comparative study of video laryngoscopy combined with bronchial blocker and video laryngoscopy combined with double-lumen tube in the teaching of endotracheal intubation in thoracic surgery in the standardized residency training of anesthesia.Methods:The trainees of the standardized residency training were randomly divided into control group and experimental group for clinical teaching, with 25 ones in each group. The experimental group was treated with visual laryngoscopy combined with bronchial blocker, while the control group was treated with visual laryngoscopy combined with double-lumen tube group. The intubation time, intubation success rate, positioning time, hemodynamic changes, and complication incidence during intubation, as well as student assessment results were recorded. GraphPad Prism 6.0 was used for t test and Chi-square test. Results:The time of endotracheal intubation [(95.3±10.1) vs. (137.5±13.5)] and positioning time [(100.8±11.7) vs. (155.4±15.3)] in the experimental group were both shorter than those of the control group ( P< 0.001), the hemodynamic changes in patients with immediate intubation were smaller ( P<0.001), the success rate of intubation was higher (92% vs. 68%) ( P<0.001), the complication incidence was lower ( P<0.001) and the students' performance was higher ( P<0.001). Conclusion:In the anesthesia teaching of thoracic surgery, bronchial blocker can reduce the time of endotracheal intubation, lower the hemodynamic changes during intubation, cut down the incidence of complications, improve the success rate of endotracheal intubation and enhance the confidence of students.

Relationship between TIPE2 and caspase-11 during pyroptosis in macrophages of mice / 中华麻醉学杂志

Objective@#To evaluate the relationship between tumor necrosis factor-alpha-induced protein 8-like 2 (TIPE2) and caspase-11 during pyroptosis in macrophages of mice.@*Methods@#J774A.1 macrophages of mice were divided into 4 groups (n=6 each) using a random number table method: non-specific siRNA (Scr-siRNA) group (S group), Scr-siRNA plus LPS/ATP group (S+ LPS/ATP group), TIPE2-siRNA group (T group) and TIPE2-siRNA plus LPS/ATP group (T+ LPS/ATP group). The corresponding siRNA and Lipofectamine2000 transfection reagents were added to each group, and transfection was performed for 24-48 h, and in addition LPS 1.0 μg/ml was then added, cells were incubated for 5 h, then ATP 5.0 mmol/L was added, and cells were incubated for 1 h in S+ LPS/ATP and T+ LPS/ATP groups.Cells were collected to detect the expression of TIPE2, caspase-11, NOD-like receptor familypyrin domain containing 3 (NLRP3) and caspase-1 (by Western blot). The supernatant was collected for determination of lactic dehydrogenase (LDH) and myeloperoxidase (MPO) activities and concentrations of interleukin-1beta (IL-1β) and IL-18 (by enzyme-linked immunosorbent assay).@*Results@#Compared with group S, the expression of TIPE2 was significantly down-regulated, the expression of caspase-11, NLRP3 and caspase-1 was up-regulated, and the activities of LDH and MPO and concentrations of IL-1β and IL-18 in supernatant were increased in group S+ LPS/ATP (P<0.05). Compared with group T, the expression of TIPE2 was significantly down-regulated, the expression of caspase-11, NLRP3 and caspase-1 was up-regulated, and the activities of LDH and MPO and concentrations of IL-1β and IL-18 in supernatant were increased in group T+ LPS/ATP (P<0.05). Compared with group S+ LPS/ATP, the expression of TIPE2 was significantly down-regulated, the expression of caspase-11, NLRP3 and caspase-1 was up-regulated, and and the activities of LDH and MPO and concentrations of IL-1β and IL-18 in supernatant were increased in group T+ LPS/ATP (P<0.05).@*Conclusion@#TIPE2 inhibits pyroptosis in macrophages through down-regulating the expression of caspase-11 in mice.

Relationship between TIPE2 and caspase-11 during pyroptosis in macrophages of mice / 中华麻醉学杂志

Objective To evaluate the relationship between tumor necrosis factor-alpha-induced protein 8-like 2 (TIPE2) and caspase-11 during pyroptosis in macrophages of mice.Methods J774A.1 macrophages of mice were divided into 4 groups (n =6 each) using a random number table method:nonspecific siRNA (Scr-siRNA) group (S group),Scr-siRNA plus LPS/ATP group (S+LPS/ATP group),TIPE2-siRNA group (T group) and TIPE2-siRNA plus LPS/ATP group (T+LPS/ATP group).The corresponding siRNA and Lipofectamine2000 transfection reagents were added to each group,and transfection was performed for 24-48 h,and in addition LPS 1.0 μg/ml was then added,cells were incubated for 5 h,then ATP 5.0 mmol/L was added,and cells were incubated for 1 h in S + LPS/ATP and T + LPS/ATP groups.Cells were collected to detect the expression of TIPE2,caspase-11,NOD-like receptor familypyrin domain containing 3 (NLRP3) and caspase-1 (by Western blot).The supernatant was collected for determination of lactic dehydrogenase (LDH) and myeloperoxidase (MPO) activities and concentrations of interleukin-1beta (IL-1β) and IL-18 (by enzyme-linked immunosorbent assay).Results Compared with group S,the expression of TIPE2 was significantly down-regulated,the expression of caspase-11,NLRP3 and caspase-1 was up-regulated,and the activities of LDH and MPO and concentrations of IL-1β and IL-18 in supernatant were increased in group S+LPS/ATP (P＜0.05).Compared with group T,the expression of TIPE2 was significantly down-regulated,the expression of caspase-11,NLRP3 and caspase-1 was up-regulated,and the activities of LDH and MPO and concentrations of IL-1β and IL-18 in supernatant were increased in group T+LPS/ATP (P＜0.05).Compared with group S+LPS/ATP,the expression of TIPE2 was significantly down-regulated,the expression of caspase-11,NLRP3 and caspase-1 was up-regulated,and and the activities of LDH and MPO and concentrations of IL-1β and IL-18 in supernatant were increased in group T+LPS/ATP (P＜0.05).Conclusion TIPE2 inhibits pyroptosis in macrophages through down-regulating the expression of caspase-11 in mice.

Effects of dexmedetomidine on NLRP3 inflammasome during acute lung injury induced by blunt chest trauma and hemorrhagic shock-resuscitation in rats / 中华麻醉学杂志

Objective To evaluate the effects of dexmedetomidine on NLRP3 inflammasome during acute lung injury ( ALI ) induced by blunt chest trauma and hemorrhagic shock-resuscitation in rats. Methods Forty-five SPF healthy male Sprague-Dawley rats, weighing 200-220 g, were divided into 3 groups (n=15 each) using a random number table method: sham operation group (Sham group), ALI group and dexmedetomidine group ( Dex group) . A rat model of ALI was established by blunt chest trauma and hemorrhagic shock-resuscitation. Dexmedetomidine 5 μg·kg-1 ·h-1 was infused into the femoral vein after blunt chest trauma in Dex group. Blood samples were collected from the femoral artery at 6 h after blunt chest trauma for measurement of arterial oxygen partial pressure and concentrations of pro-inflammatory cytokines interleukin-1beta ( IL-1β) and IL-18 in serum ( by enzyme-linked immunosorbent assay ) . The oxygenation index was calculated. The rats were sacrificed and lungs were removed for examination of the pathological changes which were scored and for determination of the expression of NLRP3, caspase-1 and apoptosis-associated speck-like protein containing CARD ( ASC) in lung tissues ( by Western blot) and ac-tivities of lactic dehydrogenase ( LDH) and myeloperoxidase ( MPO) in lung tissues ( by colorimetry) . Re-sults Compared with Sham group, PaO2 and oxygenation index were significantly decreased, the expres-sion of NLRP3, caspase-1 and ASC was up-regulated, the activities of LDH and MPO were increased, and the concentrations of IL-1β and IL-18 in serum were increased in ALI group ( P＜0. 05 ) . Compared with ALI group, PaO2 and oxygenation index were significantly decreased, the expression of NLRP3, caspase-1 and ASC was down-regulated, the activities of LDH and MPO were decreased, and the concentrations of IL-1β and IL-18 in serum were decreased in ALI group ( P＜0. 05) . Conclusion Dexmedetomidine can al-leviate blunt chest trauma and hemorrhagic shock-resuscitation induced ALI in rats, and the mechanism is related to inhibition of NLRP3 inflammasome activation and reduction of inflammatory response.

Primary culture of cat intestinal epithelial cell and construction of its cDNA library / 中国血吸虫病防治杂志

Objective To establish the primary cat intestinal epithelial cells(IECs)culture methods and construct the cD-NA library for the following yeast two-hybrid experiment,so as to screen the virulence interaction factors among the final host. Methods The primary cat IECs were cultured by the tissue cultivation and combined digestion with collagenase XI and dispase I separately. Then the cat IECs cultured was identified with the morphological observation and cyto-keratin detection ,by using goat anti-cyto-keratin monoclonal antibodies. The mRNA of cat IECs was isolated and used as the template to synthesize the first strand cDNA by SMARTTM technology,and then the double-strand cDNAs were acquired by LD-PCR,which were subsequently cloned into the plasmid PGADT7-Rec to construct yeast two-hybrid cDNA library in the yeast strain Y187 by homologous recom-bination. Matchmaker?Insert Check PCR was used to detect the size distribution of cDNA fragments after the capacity calcula-tion of the cDNA library. Results The comparison of the two cultivation methods indicated that the combined digestion of colla-genase XI and dispase I was more effective than the tissue cultivation. The cat IECs system of continuous culture was established and the cat IECs with high purity were harvested for constructing the yeast two-hybrid cDNA library. The library contained 1.1× 106 independent clones. The titer was 2.8 × 109 cfu/ml. The size of inserted fragments was among 0.5-2.0 kb. Conclusion The yeast two-hybrid cDNA library of cat IECs meets the requirements of further screen research,and this study lays the foundation of screening the Toxoplasma gondii virulence interaction factors among the cDNA libraries of its final hosts.

Prokaryotic expression and identification of rhoptry protein 38 of Toxoplas-ma gondii / 中国血吸虫病防治杂志

Objective To explore the biological function of rhoptry protein 38(ROP38)of Toxoplasma gondii,and to iden?tify the reactogenicity of the recombinant protein(rROP38). Methods The ROP38 was amplified by RT?PCR from T. gondii RH strain,and was cloned into prokaryotic expression vector pET?28a(+). The recombinant plasmid was transformed into E. co?li BL21(DE3)competent cells. Then the rROP38 was analyzed by SDS?PAGE and identified by Western blot. Results SDS?PAGE showed that rROP38 was efficient expression with a molecular weight of about 43 kD. Western blot showed that rROP38 reacted with antibody of His tag or human positive antibody,which indicated that ROP38 had good reactogenicity and could be a serological diagnostic antigen. Conclusion The study successfully obtains the rROP38 of T. gondii with good reactogenicity.

Development and application of rapid molecular method for detection of as-ymptomatic infection of Leishmania / 中国血吸虫病防治杂志

Objective To develop a rapid molecular biological method for detection of the asymptomatic infection of Leish?mania. Methods Two pairs of primers named RV1?RV2 and K13A?K13B were selected to be the fast diagnosis primers since they were designed according to the conserved region of Leishmania kinetoplast DNA(kDNA)minicircles. The PCR amplifica?tion products of Leishmania donovani promastigote from Shandong Province were sequenced to compare their conservatism. The method was applied to detect 105 venous blood samples from healthy home canine and 7 venous blood samples from home canine suffered from Kala?azar in Heishui County of Sichuan Province,and 75 venous blood samples from susceptible population(no leishmaniasis symptoms)and 7 venous blood samples from patients in Xinjiang Kashi area in order to verify the feasibility and accuracy of the method. Results The size of PCR products was consistent with the expected fragments with high conservative among Leishmania species. The positive rates of 105 home canine samples and 75 susceptible population samples were 37.14%(39/105)and 82.67%(62/75)rspectively,and the positive rates of home canine suffered from Kala?azar and patients were all 100%(7/7). Conclusion This rapid diagnosis method is suitable for detection of asymptomatic infection of Leishmania in Kala?azar endemic areas of China with high sensitive and specific,thus it has bright perspective to be used.

Cloning,prokaryotic expression and immunological identification of Toxo-plasma surface antigen IMP1 / 中国血吸虫病防治杂志

Objective To subclone express and identify the immune mapped protein 1 IMP1 which encodes a surface an?tigen of Toxoplasma gondii. Methods The cDNA of T. gondii RH strain was synthesized by reverse transcription PCR the IMP1 open reading frame ORF was amplified by PCR using the T. gondii RH strain cDNA as template the PCR products were identified by TA?cloning and sequencing then the IMP1 ORF was subcloned into the NdeⅠand Xho I sites of the vector pET28b and the positive recombinant pET28b?IMP1 was identified by double?digesting and sequencing. The protein of 6 × His tagged IMP1 was inducibly expressed in E. coli strain BL21 DE3 with isopropylβ?D?1?thiogalactopyranoside IPTG and the induction time concentration of IPTG and temperature gradients to optimize protein expression conditions were determined. After the cells carried IMP1 were induced by the optimized conditions and harvested the resulting bacteria were suspended in resuspension buffer and lysed by sonication and the supernatants were loaded onto the Ni2+Chelating Sepharose Fast Flow col?umn for affinity chromatography of the N?terminal 6 × His tagged IMP1 protein. Finally the fusion IMP1 proteins were identified by Western blotting. Results The ORF sequence of IMP1 was successfully subcloned from the cDNA of Toxoplasma Gondii RH strain and the amplified product was sequenced and identified based on which the IMP1 ORF gene was inserted into the prokaryotic expression vector pET28b and the recombinant pET28b?IMP1 was constructed successfully. The double?digesting and sequencing results indicated the validity of the recombinant vector. And the optimized conditions for the expression of IMP 1 was determined namely 0.3 mmol/L IPTG induction for 9 h at 20℃. Furthermore IMP1 protein was expressed solubly and che?lated on Ni2+sepharose beads with high affinity thus this protein could be purified efficiently by affinity chromatography. The pure fusion protein was confirmed with fine immunocompetence by SDS?PAGE and Western blotting. Conclusions IMP1 pro? tein can be high efficiently expressed by the E. coli prokaryotic expression systems the protein of IMP1 is soluble and has stable characters. The study may lay a useful foundation for the following works including in vivo expression of IMP1 crystal structure study of IMP1 and anti?toxoplasmosis subunit vaccine development.

Research progress of interaction between Toxoplasma gondii rhoptry pro-teins and host cells / 中国血吸虫病防治杂志

Rhoptry proteins are the major virulence factors of Toxoplasma gondii. They locate in different parts of the host cells and can affect the membrane cytoskeleton structure and active factors of the host cells so as to block the cell intrinsic de-fense mechanisms of the host and let T. gondii invade parasitize and proliferate in the host successfully. The function and ac-tion mode of rhoptry proteins reflect the pathogenic mechanism of T. gondii which holds great significance to looking for toxo-plasmosis drug targets and developing molecule vaccines. This paper reviews the research progess of the interaction between rhoptry proteins of T. gondii and host cells.

Therapeutic effect of 138 imported falciparum malaria patients / 中国血吸虫病防治杂志

Objectives To summarize the clinical characteristics of imported falciparum malaria patients and the treatment so as to provide the evidences for improving the diagnosis and treatment of the disease. Methods A total of 138 imported falci?parum malaria patients who received the treatment in Shandong Institute of Parasitic Diseases from January 2007 to February 2013 were adopted as the observation subjects and their clinical data were collected and analyzed. Results All the 138 pa?tients were back from African countries. The main manifestations were fever headache asthenia and hepatosplenomegaly and most of them were with decreased RBC PLT levels and increased LDH levels and 36.96%of them were misdiagnosed as respiratory diseases nephritis hepatitis and so on. Through antimalarial treatment of artemether or artesunate or dihydroartemis?inin and primaquine or dihydroartemisinin and piperaquine and symptomatic treatment the short?term and long?term cure rates were 98.55%and 94.93%respectively with 1 case unrecovered and 1 died. Conclusions Artemisinins are still the most effective antimalarial drugs for falciparum malaria. However some patients recrudesce as the Plasmodium in their body is resis?tant or insensitive to these drugs. We should pay more attention to the antimalarial and symptomatic treatments in the early stage of severe malaria so as to improve the cure rate.

Pseudokinases secreted by rhoptry in Toxoplasma gondii / 中国人兽共患病学报

Toxoplasma gondii rhoptry proteins (ROPs) include dozens of proteins secreted by tachyzoites ,these ROPs are known to be critical for several processes such as invasion ,parasitophorous vacuole establishment ,proliferation ,egress and getting out of host immune protection .ROPs belong to the protein kinase family and have classic kinase domain ,however ,most of the members are designated as pseudokinases due to the lack of critical catalytic residues that are normally required for phos-phorylation of host signal molecules .Recent studies indicated that pseudokinases also have important functions ,such as regula-ting kinase activity by allosteric and promoting the interactions between proteins as scaffolds .In this paper ,we summarize the pseudokinases structural characteristics and some pseudokinases functions ,for the study of toxop lasma gondii and host interac-tion mechanisms ,molecular vaccines screening and drugs design .

Microwave processing of Alumen / 中成药

AIM: To establish an effective and economical processing method for high quality of Calcinatum Alumen with microwave heating. METHODS: A certain amount of Alumen was placed into microwave oven and heated, and its weight was determined by electronic balance every one or five minutes. The quality (appearance, aqueous solubility, water content) of Calcined Alum by microwave was compared with those of products prepared by traditional method, far IR, and electric oven. Moreover, the stability of Calcined Alum in several kinds of package were investigated at room temperature. RESULTS: The microwave for the processing of Alumen has the advantage of rapidness, low cost, and less water content, and its product could be dissolved completely in water. CONCLUSION: Microwave is a practical technique for Alumen processing.

Microwave processing of Borax / 中成药

AIM:To establish a new processing method for Borax with microwave technique. METHODS: A certain amount of Borax was placed into microwave oven and heated, and its weight loss was determined by electronic balance. The optimum processing parameters were obtained through orthogonal design. Water content of Calcined Borax was determined by far IR, microwave oven, and electric oven(at (400 ?C)),respectively. In addition, the stability of Calcined Borax was studied at room temperature. RESULTS: Calcined Borax will be available (within) 30 min with microwave treatment. The total water content can be determined by electric oven(at (400 ?C)). Calcined Borax should be powdered and packed as soosn as possible after processing. CONCLUSION: Microwave is a kind of excellent method for preparation of Calcined Borax in laboratory, with advantages of rapidness and simplicity.

Identification of Six Trichinella Isolates from China by Multiplex PCR / 中国寄生虫学与寄生虫病杂志

Objective To identify and classify six isolates of swine-originated Trichinella from China. Methods Five specific pairs of primers were synthesized based on DNA sequence of expansion segment V region and internal tran-scribed spacers (ITS1 and ITS2) of ribosomal DNA repeat from Trichinella. International reference strains of five Trich-inella species [Trichinella spiralis (T1), T. nativa (T2), T. britovi (T3), T. pseudospiralis (T4) and T. nelsoni(T7)] were used as control. Six swine Trichienlla isolates from Henan, Yunnan, Harbin, Tongjiang of Heilongjiang, Hubei and Tianjin were identified by multiplex PCR and its effecting factors of PCR amplification were observed. Results Electrophoresis results of multiplex PCR products of Trichinella larvae showed that the band (173 bp) of the six isolates was the same as T. spiralis(T1). The specific band (173 bp) was detected by multiplex PCR through amplification from issues of single T. spiralis larva, the larvae conserved in 80% ethanol for 6 months, the larvae stored in 10% formaldehyde, in 0.05% formaldehyde, 0.2% sodium azide or 0.05% merthiotate for 2 weeks,or fresh mouse muscle with larvae. Conclusion All the six swine Trichinella isolates are identified as T. spiralis (T1) by multiplex PCR.