Objective To construct and express a recombinant plasmid pGEX-Sj26GST of Schistosoma japonicum(Sj) in Escherichia coli(E.coli) BL21 (DE3).Methods Total RNA was extracted from Sj adult worms by RNeasy Mini kit,26 kilodalton glutathione-S-transferases of Schistosomajaponicum (Sj26GST) antigen gene was amplified by real-time PCR(RT-PCR) from the total RNA,then cloned into a prokaryotic expression plasmid pGEX1λT and transformed into E.coli BL21 (DE3) to construct pGEX-Sj26GST; BL21 (pGEX-Sj26GST) was induced with isopropyl-beta-D-thiogalactopyranoside (IPTG),and the expressed products were analyzed and identified by sodium dodecyl sulfate polyacrylamide gel electrophoresis(SDS-PAGE) and Western blotting.Results The 676 bp Sj26GST gene was successfully amplified by RT-PCR and restriction enzyme double-digestion technique confirmed that Sj26GST antigen gene was successfully cloned into pGEX-1λT vector,the relative molecular mass of the expressed recombinant protein was approximately 52 × 103 by SDS-PAGE,and the amount of expressed protein was 20％ of the total bacterial proteins; the fusion protein could be recognized by sera from rabbits infected with Sj by Western blotting.Conclusions The recombinant plasmid pGEX-Sj26GST is successfully constructed and highly expressed in E.coli and the expressed fusion protein shows specific antigenicity.

This paper has reported the effect of preservation myceh'al fungus by the methods of slant and rubber plug. The mycelial fungus preserved include 29 genera, 69 species, 128 strains. The preservation time is from 2 years to 17 years. By slant inoculation, their survival conditions are still good keep their original charaters. The result shows that it is simple and effective preserving some mycelial fungus by the methods of slant and rubber plug.

OBJECTIVETo explore the protective effect and mechanism of total flavones of Bidens pilosa L. (TFB) on IgA1 induced injury of venous endothelial cells in Henoch-Schönlein purpura (HSP) children patients. METHODS Human umbilical venous endothelial cells (HUVECs) were taken as subject. They were intervened by normal IgA1 and HSP children patients' serum IgA1, and added with different concentrations TFB at the same time. Then they were divided into the blank control group, the normal control group, the HSP IgA1 group, and HSP IgA1 plus TFB (1.0, 0.5, 0.25 mg/mL) groups. Levels of TNF-α and IL-8 in supernate were detected by ELISA. The NO level was detected by nitrate reductase method. mRNA and protein expressions of NF-κB and ICAM-1 in HUVECs were detected by fluorescent quantitative PCR and Western blot respectively.

RESULTSCompared with the normal control group and the blank control group, levels of IL-8, TNF-α, and NO all significantly increased in the HSP group (P < 0.05). Compared with the HSP group, levels of IL-8, TNF-α, and NO significantly decreased after intervention of TFB (1.0 and 0.5 mg/mL; P < 0.05, P < 0.01). Results of fluorescent quantitative PCR and Western blot showed, as compared with the blank control group and the normal control group, mRNA and protein expressions of NF-κB and ICAM-1 in HSP children patients' serum IgA1 induced venous endothelial cells significantly increased with statistical difference (P < 0.05, P < 0.01). Compared with the HSP group, mRNA and protein expressions of NF-KB and ICAM-1 were obviously down-regulated after intervention of TFB (1.0, 0.5, 0.25 mg/mL), with statistical difference (P < 0.05, P < 0.01).

CONCLUSIONTFB could protect vascular damage by inhibiting in vivo high expression of NF-κB, reducing the production of IL-8, TNF-α, and NO in vascular endothelial cells of HSP children patients.

Bidens ; chemistry ; Child ; Flavones ; pharmacology ; Human Umbilical Vein Endothelial Cells ; drug effects ; Humans ; Immunoglobulin A ; blood ; Intercellular Adhesion Molecule-1 ; metabolism ; Interleukin-8 ; metabolism ; NF-kappa B ; metabolism ; Nitric Oxide ; metabolism ; Purpura, Schoenlein-Henoch ; blood ; RNA, Messenger ; metabolism ; Tumor Necrosis Factor-alpha ; metabolism

Objective To evaluate the effects of the sciatic nerve block on emergence agitation following sevoflurane-based anesthesia in pediatric patients undergoing foot orthomorphia.Methods Sixty pediatric patients,aged 3-7 yr,weighing 12-20 kg,of ASA physical status Ⅰ or Ⅱ,scheduled for elective foot orthomorphia,were randomly assigned to one of two groups (n =30 each):control group (group Ⅰ) and sciatic nerve block group (group Ⅱ).After induction of anesthesia,laryngeal mask airway was inserted,and artificial or mechanical ventilation was performed.Anesthesia was maintained with inhalation of sevoflurane (end-tidal concentration 0.5％-2.0％) and iv fentanyl 0.5-2.0 μg/kg.The sciatic nerve block on the affected side was performed under the guidance of a nerve stimulator,and 0.25％ bupivacaine 0.5 ml/kg was injected.Laryngeal mask airway removal time and emergence time were recorded.At 30 min of recovery from anesthesia,agitation was scored and the development of emergence agitation was recorded.Pain was evaluated with Faces Pain Scale-Revised (FPS-R) at 10 and 30 min of recovery from anesthesia.Adverse reactions including hypoxemia and nausea and vomiting were also recorded during recovery from anesthesia.Results Compared with group Ⅰ,the intraoperative consumption of fen tanyl and mean end-tidal concentration of sevoflurane were significantly decreased,laryngeal mask airway removal time and emergence time were shortened,and the incidence of emergence agitation,PPS-P scores and incidence of hypoxemia were decreased in group Ⅱ.Conclusion The sciatic nerve block is helpful in decreasing the development of emergence agitation following sevoflurane-based anesthesia in pediatric patients undergoing foot orthomorphia.

Bacillus cereus B-04 antagonist to Botrytis cinerea were isolated from samples of tomato soil infected by Botrytis cinerea in Zibo, which are identified through a series of morphological and biochemical characteristics and the sequence of 16SrDNA. Aiming at enhancing the inhibitory effect of this strain, a 4.1kb DNA fragment containing ?-1,3-glucanases gene from pUC1940 was inserted into vector pBE2 and pHY300PLK to construct recombination plasmids, PBE2-glu and pHY300PLK-glu, which were transferred into Bacillus cereus B-04, resulting in a new strain named B-04-glu. Restriction enzyme digestion and ?-1,3-glucanases plate culture confirmed that B-04-glu contained a functional ?-1,3-glucanases gene. Compared to the wild strain B-04, B-04-glu had an increased inhibitory effect against Botrytis cinerea on tomato.

Objective To explore the relationship between levels of serum 25-(OH) D3 and changes in Th1/Th2 cell balance in infants with recurrent wheezing.Methods Sixty cases of infants with recurrent wheezing were involved and 60 cases of healthy children were selected as controls.Enzyme linked immunosorbent assay (ELISA) was used to detect the levels of serum 25-(OH) D3 and double-antibody sandwich (ABC-ELISA) was used to detect the serum levels of interferon-γ(IFN-γ),interleukin (IL)-4,IL-13,and then the relationship between the levels of serum 25-(OH) D3 and changes in Th1/Th2 balance in infants with recurrent wheezing were explored.Results Serum 25-(OH) D3 levels decreased significantly in the infants with recurrent wheezing group compared with those of the healthy control group [(18.24 ± 5.64) μg/L vs (37.85 ± 7.78) μg/L] (t =15.810,P =0.000).Serum IFN-γ levels decreased significantly in the infants with recurrent wheezing group compared with those of the healthy control group [(11.20 ± 2.08) ng/L vs (20.68 ± 3.87) ng/L] (t =16.700,P =0.000).In contrast,serum IL-4,IL-13 levels increased significantly in the infants with recurrent wheezing group compared with those of the healthy control group[IL-4:(28.61 ±6.44) ng/L vs (22.14±5.29) ng/L;IL-13:(20.02±4.83) ng/L vs (17.72± 4.06) ng/L] (t =6.201,P =0.000 ; t =2.829,P =0.006).Th1/Th2 in the infants with recurrent wheezing group were lower than that those of the healthy control group,and there was statistically significant difference between two groups(0.41 ± 0.12 vs 1.00 ± 0.36) (t =11.796,P =0.000).Serum 25-(OH) D3 levels were negatively correlated with Th1/Th2 in the infants with recurrent wheezing(r =-0.649,P =0.000).There were no correlation between serum 25-(OH) D3 levels and Thl/Th2 in the healthy control group(r =-0.217,P =0.096).Conclusions Low serum 25-(OH) D3 may be the risk factor for recurrent wheezing in infants.Serum 25-(OH) D3 levels were negatively correlated with Th1/Th2 in the infants with recurrent wheezing group,which show that recurrent wheezing in the infants is closely related to allergic reaction.

Coronavirus disease 2019 (COVID-19) is a highly contagious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). More than one-third of patients with COVID-19 experience neurological symptoms, including confusion, headaches, and decreased/disordered taste. Alzheimer's disease (AD) is a slowly progressive neurodegenerative disease and the most common type of dementia. Alzheimer's disease patients are at high risk and susceptible to infection with COVID-19, which may cause severe illness and even death. There appears to be an interaction between AD and COVID-19, and on the one hand, patients with COVID-19 seem to be more likely to develop AD. AD patients, on the other hand, may be more susceptible to severe COVID-19. Therefore, understanding the common link between COVID-19 and AD may help to develop treatment strategies. Risk factors common to AD and COVID-19 are aging, ApoE ε4 allele, β-amyloid (Aβ) deposition, angiotensin-converting enzyme (ACE), neuroinflammation, oxidative stress. Here, this article focuses on the relationship between COVID-19 and AD, explores common risk factors and potential pathogenesis, and provides help for early prevention, treatment and recovery.

Objective To observe effects and mechanisms of insulin on reperfusion injury after cerebral ischemia.Methods Sixty-six male Wistar rats were used in this study.All rats were divided into 3 groups as treated group(A),control group(B) and random sham-operated group(C).Four-vessel occlusion was used to establish global cerebral ischemia reperfusion model in study groups.The treated group were divided into 5 groups(A1-A5) and intraperineally injected with biosynthetic human insulin 2 IU/(kg?d) and 50%glucose 2 g/(kg?d) for 7 days,the blood glucose was monitored in preoperative and postoperative 3,6,12,24 h,and the blood glucose was maintained between 3.5-6.5 mmol/L.These animals of control group were given with saline 2 mL/(kg?d) for 7 days in abdominal cavity.All the rats were killed in the seventh day,brain homogenate was collected for detection of neuron specific enolase(NSE)and nitric oxide(NO).The hippocampus was separated for observation of electronic microscope.Results Concentration of NSE in brain tissue in group C was significantly higher than that of group A and group B,while the level in group A was higher than that of group B.Concentration of NO in group C was lower than that of group A and group B while the level of NO in group A was significantly lower than that of group B.Electron microscope showed that the ultrastructure of sham-operated group was nearly normal,damage degree of hippocampal neuron and gliacyte and capillary was gradually worse from group A1,A2 to A4,the damage degree of group B1,B2 and B4 was serious and there was no difference among them.Conclusion Insulin can really promote recovery of the cerebral injury after ischemia reperfusion.

Diathesis education and innovative ability cultivating of students are a new position in higher education.Exploring experiment was applied in medical undergraduates of grade 2004 for enhancing integrative diathesis and cultivating innovative spirit and ability of students.It was proved that application of exploring experiment could increase adequately learning fervor and interest of students.Teaching quality of microbiological experiment was enhanced accordingly.Application of exploring experiment was useful supplement and attempt of traditional teaching mode.

Objective To investigate the expression of the cyclin D1 in pubertal mammary hypertrophy and micromastia. Methods The expression of cyclin D1 was detected by SP immunohisto-chemistry in 18 cases of pubertal mammary hypertrophy and 12 cases of micromastia. Results The expression rate of cyclin D1 within breast tissue was 83. 33 % and 16. 67 % in pubertal mammary hypertrophy and micromastia, respectively. There was significant difference between the two groups (P<0. 01). Conclusion The expression level of the cyclin D1 within breast tissue in pubertal mammary hypertrophy is higher than that in micromastia.