In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sj14 or pVAC-Sj26 only to get two gene fragments including Sj14 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES, resulting in another new recombinant plasmid pIRES-Sj26-Sj14. The expression of Sj14 and Sj26 genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and plRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

In order to construct a eukaryotic co-expression plasmid containing membrane-anchored Sjcl4FABP and Sjc26GST genes and identify their expression in vitro, Sj14 and Sj26 genes were obtained by RT-PCR with total RNA of Schistosoma japonicum adult worms as the template and cloned into eukaryotic expression plasmid pVAC to construct recombinant plasmids pVAC-Sj14 and pVAC-Sj26. Then a 23 amino-acid signal peptide of human interleukin-2 (IL-2) upstream Sj14 or Sj26 gene and a membrane-anchored sequence containing 32 amino-acids of carboxyl-terminal of human placental alkaline phosphatase (PLAP) downstream were amplified by PCR as the template of plasmid pVAC-Sjl4 or pVAC-Sj26 only to get two gene fragments including Sjl4 gene and Sj26 gene. The two modified genes were altogether cloned into a eukaryotic co-expression plasmid pIRES,resulting in another new recombinant plasmid pIRES-Sj26-Sj 14. The expression of Sj14 and Sj26genes was detected by RT-PCR and indirect immunofluorescent assays (IFA) when the plasmid pIRES-Sj26-Sj 14 was transfected into eukaryotic Hela cells. Restriction enzyme analysis, PCR and sequencing results revealed that the recombinant plasmids pVAC-Sj14, pVAC-Sj26 and pIRES-Sj26-Sj14 were successfully constructed and the expression of modified Sj 14 and Sj26 genes could be detected by RT-PCR and IFA. A bivalent membrane-anchored DNA vaccine encoding Sj14 and Sj26 genes was acquired and expressed proteins were proved to be mostly anchored in cellular membranes.

To find a new preventive strategy for the infection of Schistosoma japonica, plasmid pIRES-Sj97-Sj14-Sj26 that contains fatty binding protein (Sj14), GST (Sj26) and paramyocin (Sj97) that are expressed on the membrane, was constructed. RT-PCR was used to detect the expression of Sj14 mRNA, Sj26 mRNA and Sj97 mRNA in the Hela cells, the indirect immunofluorescent test was employed for the detection of the expression of trans-membrane Sj26 after the plasmid was trans-fected into Hela cells. Fifty BALB/c mice were randomly divided into 5 groups and pIRES-Sj97-Sj14-Sj26 plasmid DNA, pIRES-Sj14-Sj26 plasmid DNA, plRES-Sj26 plasmid DNA,plRES blank vector and normal saline were respectively injected into the quadriceps muscles of thigh.Eight weeks after the immunization the mice were killed and significantly higher level of IgG was detected in the pIRES-Sj97-Sj14-Sj26 group as compared with the plRES blank vector, normal saline and pIRES-Sj26 groups (P<0.01) and the pIRES-Sj14-Sj26(P<0.05). Single splenocyte suspension was prepared to detected the level of IFN-γ by ELISA and the lymphocyte stimulating index (SI) by MTT SI was significantly higher of in the pIRES-Sj97-Sj14-Sj26 group than in other groups (P<0.01), while the IFN-γ, level was significantly higher the pIRES-Sj97-Sj14-Sj26 group than in pIRES blank vector and normal saline groups (P<0.01), but no significant differences were found when compared with pIRES-Sj14-Sj26 and pIRES-Sj26 groups. Flow cytometery showed that the percent-ages of CD4+ and CD8+ T cells were much higher in the pIRES-Sj97-Sj14-Sj26 group (P<0.01,P<0.05). It was concluded that pIRES-Sj97-Sj14-Sj26 vaccine may induce stronger immune response in BALB/c mice.

Objective: To investigate the influencing factors of fertility outcome after laparoscopic conservative surgery for tubal pregnancy. Methods: From October 2010 to October 2016, 253 cases of tubal pregnancy treated by laparoscopic conservative surgery in General Hospital of Jizhong energy Fengfeng Group Hospital were analyzed retrospectively.All patients were followed up from 24 to 36 months after operation to observe the intrauterine pregnancy.Logistic regression was used to analyze the influencing factors of intrauterine pregnancy. Results: After 24-36 months follow-up, the patients were not contraception and pregnant under the guidance of doctors.Among the 253 cases, 182 (71.1%) were intrauterine pregnancy, 37 (14.6%) were ectopic pregnancy, and 34 (13.4%) were not pregnant.The results of logistic regression showed that high level of hCG, severe pelvic adhesions, obstruction of fallopian tube and history of ectopic pregnancy were the risk factors of intrauterine pregnancy (OR (95%CI) 1.982 (1.075-3.149), 2.410 (1.279-5.069), 2.485 (1.071-3.594), 5.071 (1.094-9.081), P<0.05 or P<0.01). Conclusion The reproductive outcome of laparoscopic conservative surgery for tubal pregnancy is influenced by many factors.The high level of hCG in preoperative blood, severe pelvic adhesions, obstruction of tubal and ectopic pregnancy history are the risk factors of postoperative pregnancy.

Objective:To implement an intervention with community-dwelling middle-aged and elderly people through a new science promotion approach, and to evaluate changes in the level of bone health awareness.Methods:From April 2017 to June 2017, 200 community-dwelling middle-aged and elderly people aged 50 years and over were randomly included.They were divided into the intervention group receiving a new science promotion intervention(n=100, with 50 males and 50 females)and the control group(n=100, with 50 males and 50 females). Baseline and follow-up assessments of bone health awareness were performed before and after the one-year intervention.A questionnaire survey on preferences for different types of science promotion articles in WeChat accounts was conducted.Differences in scores for intervention effects before and after intervention were compared between the two groups.Potential influencing factors for the scale score were examined by a multi-factor analysis.Results:After one year of intervention, scores for intervention effects were higher in the intervention group than in the control group( P<0.001). In the intervention group, scores were higher after intervention than before intervention( P<0.001). In the control group, scores after follow-up were slightly higher than those at baseline(29.4±11.4 vs.27.9±10.1, P<0.001). The increase in scores before and after intervention was greater in the intervention group than in the control group( P<0.001). Baseline score, group designation, history of drinking and diabetes impacted the scores in middle-aged and elderly women, while other factors did not.Most middle-aged and older people preferred texts illustrated with pictures, and older people were more receptive to videos. Conclusions:The new science promotion method is beneficial to bone health management, can improve bone health awareness in middle-aged and elderly people.

Heat shock protein 65 (HSP65) is one of the most important protective immunogens against the tuberculosis infection. The signal sequence of antigen 85B and the whole HSP65 DNA sequence of human Mycobacterium tuberculosis (M. tuberculosis) were amplified from BCG genome and plasmid pCMV-MTHSP65 respectively by polymerase chain reactions (PCR). These two sequences were cloned into the plasmid pBCG-2100 under the control of the promoter of heat shock protein 70 (HSP70) from human M. tuberculosis, yielding the prokaryotic shuttle expression plasmid pBCG-SP-HSP65. Results of restriction endonuclease analysis, PCR detection and DNA sequencing analysis showed that the two cloned DNA sequences were consistent with those previously reported, and the direction of their inserting into the recombinant was correct and the reading frame had been maintained. The recombinants were electroporated into BCG to construct the recombinant BCG vaccine and induced by heating. The induced expression detected by SDS-PAGE showed that the content of 65 kD protein expressed in recombinant BCG was 35.69% in total bacterial protein and 74.09% in the cell lysate supernatants, suggesting that the recombinant HSP65 gene could express in BCG with high efficiency and the expressed proteins were mainly soluble. Western-blot showed that the secretive recombinant proteins could specifically combine with antibody against M. tuberculosis HSP65, indicating that the recombinant proteins possess the biological activity of HSP65.
BCG Vaccine ; biosynthesis ; immunology ; Bacterial Proteins ; biosynthesis ; immunology ; Chaperonin 60 ; Chaperonins ; biosynthesis ; immunology ; Cloning, Molecular ; Escherichia coli ; metabolism ; Genetic Vectors ; Humans ; Mycobacterium tuberculosis ; genetics ; immunology ; Plasmids ; genetics ; Sequence Analysis, DNA ; Vaccines, Synthetic ; biosynthesis ; immunology

Cardiac arrest is the fourth stage of sudden cardiac death, which is characterized by the cessation of electrical activity in the heart, rapid circulatory and respiratory failure, and the prognosis is often poor. How to effectively predict cardiac arrest is the key and difficult point in the diagnosis and treatment process. In recent years, the research on the application of early warning scoring system in cardiac arrest has made continuous breakthroughs, from initially formulating a traditional scoring system containing only basic vital signs indicators according to a certain number of samples to continuously increasing and changing indicators, increasing the sample size, and formulating an improved scoring system with better sensitivity and specificity. Nowadays, with the continuous development of electronic information technology, machine learning technology is introduced into the formulation of scoring system, which breaks through the limitations of previous scoring system and has achieved good results in clinic. This article analyzes and compares the relevant research and cutting-edge progress of different early warning scoring systems at home and abroad, and summarizes the research results, gaps and shortcomings. Finally, combined with the relevant policies of graded diagnosis and treatment in China, this paper discusses the development and application direction of cardiac arrest early warning scoring system in the future.

Aristolochic acid (AA), extracted from Aristolochiaceae plants, plays an essential role in traditional herbal medicines and is used for different diseases. However, AA has been found to be nephrotoxic and is known to cause aristolochic acid nephropathy (AAN).AA-induced acute kidney injury (AKI) is a syndrome in AAN with a high morbidity that manifests mitochondrial damage as a key part of its pathological progression. Melatonin primarily serves as a mitochondria-targeted antioxidant. However, its mitochondrial protective role in AA-induced AKI is barely reported. In this study, mice were administrated 2.5 mg/kg AA to induce AKI. Melatonin reduced the increase in Upro and Scr and attenuated the necrosis and atrophy of renal proximal tubules in mice exposed to AA. Melatonin suppressed ROS generation, MDA levels and iNOS expression and increased SOD activities in vivo and in vitro. Intriguingly, the in vivo study revealed that melatonin decreased mitochondrial fragmentation in renal proximal tubular cells and increased ATP levels in kidney tissues in response to AA. In vitro, melatonin restored the mitochondrial membrane potential (MMP) in NRK-52E and HK-2 cells and led to an elevation in ATP levels. Confocal immunofluorescence data showed that puncta containing Mito-tracker and GFP-LC3A/B were reduced, thereby impeding the mitophagy of tubular epithelial cells. Furthermore, melatonin decreased LC3A/B-II expression and increased p62 expression. The apoptosis of tubular epithelial cells induced by AA was decreased. Therefore, our findings revealed that melatonin could prevent AA-induced AKI by attenuating mitochondrial damage, which may provide a potential therapeutic method for renal AA toxicity.

Objective:To study the safety and short-term clinical effect of no-touch technique for harvesting great saphenous vein in off-pump coronary artery bypass grafting(OPCABG).Methods:The clinical data of 117 patients in the single medical group who underwent OPCABG from December 2018 to April 2020 were analyzed retrospectively, including 55 patients in the no touch group (NTSV group) and 62 patients in the traditional open access group (traditional SV group). To evaluate the feasibility, safety and short-term clinical effect of using no-touch technique to harvest great saphenous vein for OPCABG. The no-touch technique was mainly based on the invention of Dr. Souza’s team in Sweden. For the treatment of isolated great saphenous veins, some improvements were made according to the clinical experience of the application of sequential bridging and radial artery in our center.Results:There were no significant differences between the two groups in hypertension, diabetes, hyperlipidemia, history of myocardial infarction, ventricular aneurysm formation, left ventricular systolic function reduction, history of cerebral infarction, chronic obstructive pulmonary disease and other complications ( P>0.05). The NTSV group had higher male proportion (96.4% vs. 61.3%) and lower mean age (53.49 years old vs. 63.76 years old), and the difference were statistically significant ( P<0.01). The patients in the two groups successfully completed the operation without cardiopulmonary bypass, and the proximal anastomosis was the first. There were no statistical significances in the average number of bypass, time to acquire great saphenous vein, operation time, 24 h postoperative thoracic drainage, blood bank transfusion, secondary thoracotomy and hemostasis, perioperative myocardial infarction, hospital death, ICU stay time, postoperative hospital stay time, and poor healing of lower limb incision of the two groups. One year after the operation, 36 patients were followed up, including 15 patients in NTSV group, and 21 patients in traditional SV group. There was no recurrence of angina pectoris, no out of hospital death and other major adverse cardiovascular events. Among them, 26 patients completed CTA reexamination of coronary artery, including 12 patients in NTSV group and 14 patients in traditional SV group, the patency rate of vein bridge was 96.4% vs. 87.2%. There was no reduction of left ventricular ejection fraction and no revascularization. Conclusion:The method of using no-touch technique to harvest great saphenous vein is safe and feasible. It does not increase the operation time and the incidence of poor healing of lower limb incision. The short-term effect of applying no-touch technique for harvesting great saphenous vein to OPCABG is better than the traditional open way.

In the paper, the study was reviewed on the infrared temperature characteristics of acupoints in recent 10 years. CNKI, WANFANG, VIP, SciVerse ScienceDirect and Springer databases were retrieved, with"infrared thermal imaging" and "acupoint" as the key words. The retrieving time was from January 1, 2006 to December 31, 2016. Totally, 468 relevant papers were searched and 169 papers of them were read carefully on acupoint infrared temperature. In terms of physiological condition, pathological condition and the stimulation methods such as acupuncture, moxibustion,, embedding therapy and cupping therapy, the general situation was reviewed on the infrared temperature characteristics of acupoints separately. It was found that the study on infrared temperature characteristics of acupoints in physiological condition was limited and the characteristics discovered were not enough to systematically review the physiological and physical properties of acupoints. The study in terms of the pathological condition objectively reflected the effects and rules of diseases. It was showed in the study of acupoint infrared temperature characteristics after stimulation that the changes of infrared thermal imaging tempe-rature at some specific region induced by different therapies and parameters might be used to deduce the potential mechanism and optimal parameters or schemes of intervention method and contributed to the formation and deve-lopment of quantitative diagnosis and treatment. The authors believe that the study on infrared temperature characteristics of acupoint provides the active significance in the exploration on the physiological and physical characteristics of acupoint, the effects and rules of diseases as well as the quantitative diagnosis and treatment.