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Objective:To summarize the clinical experience of treating patients with severe acute organic fluorine poisoning using extracorporeal membrane oxygenation (ECMO).Methods:In January 2021, an acute mass organic fluorine gas poisoning incident occurred in Quzhou, Zhejiang Province. The clinical data of 4 severe patients with acute poisoning of organic fluorine treated by ECMO in our hospital were retrospectively analyzed, and the epidemiological characteristics, clinical symptoms, signs, the abnormal laboratory studies/examinations, and treatments of this kind poisoning patients, especially, the treatment pattern, support time, complications, and outcomes of ECMO were collected and analyzed.Results:All the 4 patients were male, with an average age of (52±9) years, and all of them came to the emergency department complaining chest tightness, cough and pharyngeal discomfort 6 h after exposure by inhalation. The patient’s condition progressed rapidly with severe acute respiratory failure and circulatory failure as the prominent manifestations. The mechanical ventilations were performed (13.0±4.8) h after poisoning, and ECMO treatment was performed (15.5±5.3) h after poisoning. Among them, 2 patients were treated using venoarterial (VA) ECMO, and 2 patients using venovenous (VV) ECMO, but 1 patient was converted to VA-ECMO 8 h later. The duration of ECMO support for the patients was (8.8±3.6) d. The duration of mechanical ventilation was (23.0±28.7) d and stay in intensive care unit was (42.0±55.4) d. Among them, one patient was transferred to a specialized rehabilitation hospital after the amputation surgery due to lower limb necrosis after VA-ECMO support, and the remaining 3 patients were discharged after recovery.Conclusions:ECMO support might have the irreplaceable value in the treatment of patients with severe acute organic fluorine poisoning, and should be considered as one of the reserves of regional health care system in dealing with public health emergencies.
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Objective:To investigate the relationship between adenosine diphosphate (ADP) pathway-induced platelet dysfunction monitoring by thrombelastography with hospital mortality in patients with traumatic brain injury.Methods:The clinical data of 180 patients with traumatic brain injury in Zhejiang Quhua Hospital from January 2016 to December 2018 were retrospectively analyzed. The patients underwent thrombelastography examination. Among them, the ADP pathway-induced platelet inhibition rate (ADP inhibition rate) ≤ 60% was in 74 cases (non-ADP dysfunction group), and ADP inhibition rate > 60% was in 106 cases (ADP dysfunction group). Multiple Logistic regression analysis was used to analyze the independent influencing factors of patients′ hospital mortality. Logistic regression model was used to analyze the threshold of ADP inhibition rate to predict hospital mortality.Results:There were no statistical differences in the gender composition, age, prothrombin time, international standardized ratio, activated partial thromboplastin time, platelet count, systolic blood pressure, pulse, admission to thrombelastography examination time, Glasgow coma score, trauma severity score, simple trauma grading criteria and using of antiplatelet drugs before admission between 2 groups ( P>0.05). The intubation rate and in-hospital mortality in ADP dysfunction group were significantly higher than those in non-ADP dysfunction group: 69.8% (74/106) vs. 37.8% (28/74) and 32.1% (34/106) vs. 8.1% (6/74), and there were statistical differences ( P<0.01 or <0.05). The maximum amplitude and G value in ADP dysfunction group were significantly lower than those in non-ADP dysfunction group: (61 ± 9) mm vs. (65 ± 6) mm and (9 ± 4) kD/cm2 vs. (11 ± 3) kD/cm2, the ADP inhibition rate and arachidonic acid pathway-induced platelet inhibition rate were significantly higher than those in non-ADP dysfunction group: (76 ± 22)% vs. (45 ± 18)% and (75 ± 28)% vs. (35 ± 22)%, and there were statistical differences ( P<0.05). There were no statistical difference in the reaction time, blood clot formation time and angle between 2 groups ( P>0.05). Multiple Logistic regression analysis result showed that ADP inhibition rate >60% and trauma severity score were independent predictors of increased hospital mortality in patients with traumatic brain injury ( OR = 6.21 and 1.13, 95% CI 1.21 to 31.27 and 1.05 to 1.22, P<0.05). Logistic regression model analysis result showed that ADP inhibition rate >60% was the threshold for predicting the hospital mortality rate ( OR = 6.18, 95% CI 1.2 to 33.3). Conclusions:ADP inhibition rate of thrombelastography is related to the hospital mortality in patients with traumatic brain injury.
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Objective To detect Pneumocystis carinii (Pc) DNA by loop-mediated isothermal amplification (LAMP). Methods After injected with hydrocortisone acetate for 8 weeks, the bronchoalveolar lavage fluid (BALF) of Wistar rats were collected and a portion of BALF were examined for identifying Pc organisms using microscope. Then Pc DNA was extracted by phenol-chloroform extraction. Four primers which recognized 6 distinct regions on the mtrRNA gene of Pc were designed and used for LAMP assay. To evaluate the specificity of the assay, M. tuberculosis, M. pneumoniae, C. pneumoniae, P. gondii and rat leucocyte were used as negative controls. To compare the sensitivity of the LAMP to that of conventional PCR, Pc DNA were 10-fold serially diluted and was amplified by LAMP and PCR. LAMP results were judged by naked eye, electrophoretic analysis and restriction digestion. Results Pc organisms were detected from BALF of rats injected with hydrocortisone acetate. After LAMP reaction, positive signal was observef rats injected with hydrocortisone acetate. By contrast, no positive signal was observed for the negative controls in the study. The amplified product digested by restriction enzyme demonstrated 3 bands (82, 135, 189 lip) upon agarose gel electrophoresis, in good agreement with the predicted sizes. The detection limit of LAMP assay was 1 pg/μl of Pc DNA per reaction and that of PCR was 10 pg/μ1 of Pc DNA per reaction. Conclusion LAMP assay has usefulness for rapid detection of Pc.
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A recombinant plasmid containing cathepsin B endopeptidase of Schistosoma japonicum(Sjcb2)was constructed,indentified by PCR,restrictive enzyme,digestion and DNA sequencing,and expressed into mammalian cells.Immunochemistry examination showed that the Sjcb2 gene can be expressed in the eukaryotic system,providing a basis for the development of schistosome DNA vaccine.
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Objective To express P30 surface antigen of RH strain of Toxoplasma gondii in E.coli BL21(DE3). Methods The P30 gene from Toxoplasma gondii was cloned to the pET28b vector after PCR, and the recombinant expression plasmid pET28b-P30 was constructed. Then the recombinants were transformed into E.coli BL21(DE3) after identified by the restriction enzyme digestion, PCR and DNA sequence determination annlysis. A single colony of E.coli BL21(DE3) containing the recombinant plasmid, pET28b-P30 was inoculated in LB culture, then diluted 1∶100 into 2 ml LB culture and induced by 0.2 mmol/L IPTG, and the expression product was identified by SDS-PAGE and Western blot. Results The recombinant plasmid of pET28b-P30 was constructed. ② Plasmid pET28b-P30 could express a specific 30 kDa fusion protein in E.coli BL21(DE3). Conclusions The expression plasmid which contains the gene fragment encoding P30 surface antigen of Toxoplasma gondii has been successfully constructed and is highly expressed in E.coli BL21(DE3) as an inclusion body.
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Objective To establish a method for detecting cercaria of Schistosoma japonicum.Methods The loop-mediated isothermal amplification(LAMP)was used.The cercaria DNA of Schistosoma japonicum was extracted by using GeneReleaser.Four primers which recognized 6 distinct regions on the calcium-binding protein gene of cercaria of Schistosoma japonicum were designed and used for LAMP assay and Clonorchis sinensis was used as the negative control for evaluating the specificity and 20,10,5,1 cercariae of Schistosoma japonicum were amplified by LAMP for evaluating the sensitivity.The LAMP results were judged with the naked eyes and electrophoretic analysis.Results After LAMP reaction,the positive signal was observed with cercariae of Schistosoma japonicum.By contrast,no positive signal was observed for Clonorchis sinensis.The detection limit of LAMP assay was 1 cercaria of Schistosoma japonicum per reaction.Conclusion LAMP assay has usefulness for rapid detection of cercariae of Schistosoma japonicum.
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Objective To clone and sequence the partial gene of Schistosoma japonicum phosphoglycerate kinase (SjPGK). Methods A pair of primers were designed and synthesized according to the cDNA sequence of Schistosoma mansoni phosphoglycerate kinase (SmPGK) gene. The gene fragment of SjPGK was amplified and isolated from the total RNA of S.japonicum by reverse-transcription polymerase chain reaction (RT-PCR). The gene fragment was cloned into the cloning vector of pMD18-T, The positive clones were acquired and identified with restrictive enzymes and PCR amplification. After being sequenced with DNA auto-sequence analysis instrument,the cDNA sequence of SjPGK was searched for homologue identity with NCBI BLAST program. Results The gene encoding SjPGK was obtained and isolated by RT-PCR .The fragment of SjPGK was about 830 bp.The cDNA sequence of the phosphoglycerate kinase was highly homologous between Schistosoma mansoni and Schistosoma japonicum. The identity of nucleotide sequence was 85% and score 672, and the identity of amino sequence was 94% and score 473. Conclusion The partial gene of encoding SjPGK is cloned into the cloning vector of pMD18-T, which gives the basis for discovering new candidate vaccine molecular for schistosomiasis.
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Subjective To acquire and analyze adult stage Schistosoma japonicum (Chinese strain) expressed sequence tags and new genes from an adult S. japonicum cDNA library, and to search new vaccine candidates and drug targets. Methods A cDNA library was constructed from adult stage S. japonicum. Clones were selected randomly from the cDNA library and were se-quenced. ESTs and new genes were acquired after analysis in GenBank databases by BLAST and other programs. All ESTs and new genes were submitted to GenBank and received accession numbers. Results 149 ESTs were acquired from a total 382 clones that were randomly selected from the adult S. japonicum cDNA library. All ESTs were successfully submitted to the dbEST at Genbank. Some of them were homologous with sequences of male, female, egg, schistosomula, cercaria and miracidia of S. japonicum. 18 new genes of adult S. japonicum were acquired. Some genes were housekeeping genes and some genes might be interesting as vaccine candidates or drugs targets. Conclusions The EST straltegy is a rapid, efficient and economical method to acquire ESTs and to discover new genes of adult stage S. japonicum from cDNA libraries.[
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Objective To detect Toxoplasma gondii DNA by loop-mediated isothermal amplification(LAMP). Methods DNA was extracted by phenol-chloroform extraction from T. gondii tachyzoites. Four primers which recognized 6 distinct regions on the B1 gene of T. gondii were designed and used for LAMP assay. To evaluate the specificity of the method, Plasmodium vivax, P. falciparum, Pneumocystis carinii, Schistosoma japonicum, and mouse leucocytes were used as controls. The parasite extract (T. gondii) was 10-fold serially diluted for evaluating the sensitivity of LAMP, and was amplified by LAMP. LAMP results were read with naked eye and analyzed by electrophoresis. Results After LAMP reaction, positive amplification was observed with T. gondii, but no positive signal was toted for the negative controls in the study. The sensitivity of LAMP assay reached up to 2-3 T. gondii tachyzoites/ml per reaction. Conclusion LAMP assay shows proper specificity and sensitivity for the detection of T. gondii.