Bioinformatics analysis and prokaryotic expression of Strongyloides stercoralis serine protease inhibitor 1 / 中国血吸虫病防治杂志

Objective To predict the structure and antigenic epitope of the Strongyloides stercoralis serine protease inhibitor 1 (Ss-SRPN-1) protein using bioinformatics tools, and to construct prokaryotic expression plasmids for expression of recombinant Ss-SRPN-1 protein, so as to provide the basis for unraveling the function of the Ss-SRPN-1 protein. Methods The amino acid sequence of the Ss-SRPN-1 protein was downloaded from the NCBI database, and the physicochemical properties, structure and antigenic epitopes of the Ss-SRPN-1 protein were predicted using bioinformatics tools, including ExPASy, SWISS-MODEL and Protean. Primers were designed according to the nucleotide sequences of Ss-SRPN-1, and the Ss-SRPN-1 gene was amplified, cloned and sequenced with genomic DNA extracted from the infective third-stage larvae of S. stercoralis as a template. The Ss-SRPN-1 protein sequence was cloned into the pET28a (+) expression vector and transformed into Escherichia coli BL21 (DE) cells for induction of the recombinant Ss-SRPN-1 protein expression. The recombinant Ss-SRPN-1 protein was then purified and identified using Western blotting and mass spectrometry. Results Bioinformatics analysis showed that the Ss-SRPN-1 protein, which was composed of 372 amino acids and had a molecular formula of C1948H3046N488O575S16, was a stable hydrophilic protein, and the subcellular localization of the protein was predicted to be extracellular. The Ss-SRPN-1 protein was predicted to contain 11 dominant B-cell antigenic epitopes and 20 T-cell antigenic epitopes. The Ss-SRPN-1 gene with a length of 1 119 bp was successfully amplified, and the recombinant plasmid pET28a (+)/Ss-SRPN-1 was constructed and transformed into E. coli BL21(DE) cells. The expressed recombinant Ss-SRPN-1 protein had a molecular weight of approximately 43 kDa, and was characterized as a Ss-SRPN-1 protein. Conclusions The recombinant Ss-SRPN-1 protein has been expressed successfully, and this recombinant protein may be a potential vaccine candidate against strongyloidiasis.

Diagnosis, treatment and prevention of biliary hemorrhage after percutaneous transhepatic choledochoscopic lithotomy / 中国医师杂志

Objective:To explore the diagnosis, treatment and prevention of biliary hemorrhage after percutaneous transhepatic choledochoscopic lithotomy(PTCSL).Methods:Retrospective summary and analysis were made on 48 cases of PTCSL completed by the hepatobiliary surgery department of Hunan Provincial People′s Hospital (the First Affiliated Hospital of Hunan Normal University) from January 2016 to December 2020.Results:Biliary hemorrhage occurred in 3 cases after operation, of which 1 case was considered to have a small amount of biliary mucosa bleeding, and the bleeding was stopped after blood transfusion, hemostatic agents and appropriate fluid rehydration. In the other case, a small amount of blood oozing around the T-tube sinus canal was accompanied by a small amount of bloody fluid in the T-tube, which was considered to cause arterioles or venules accompanied by bile duct bleeding during sinus expansion during operation, and the bleeding stopped after compression. Another patient underwent a short time drainage of hemorrhagic fluid ≥100 ml in T tube 15 days after operation, and was prepared for interventional therapy while receiving blood transfusion products and conservative treatment. Celiac arteriography was performed, and pseudoaneurysm was found in the right anterior lobe of the liver. Embolization was performed later and the patient discharged 10 days after embolization.Conclusions:Biliary hemorrhage is one of the more common complications after PTCSL. Increasing the understanding of this complication provides a guarantee for the safety of surgery and has clinical significance for patients with accelerated postoperative recovery.

Clinical characteristics and outcomes of adult critically ill patients with COVID-19 in Honghu, Hubei Province / 南方医科大学学报

OBJECTIVE@#To explore the clinical characteristics and outcomes of adult critically ill patients with COVID-19 and identify the risk factors correlated with in-hospital deaths.@*METHODS@#This study was conducted among 20 confirmed adult cases of COVID-19 in the Intensive Care Unit (ICU) of Honghu People's Hospital in Jingzhou City, Hubei Province. According to the final outcome, the patients were divided into survivor group and death group with 10 patients each. The demographic data, clinical manifestations and signs, laboratory findings, treatment measures and clinical outcomes were obtained from electronic medical records to compare the clinical characteristics and outcomes between the two groups. Univariate logistic analysis was used to analyze the risk factors associated with in-hospital death.@*RESULTS@#The mean age of patients with confirmed COVID-19 was 70 ± 12 years, and 40% of them were male. The patients were admitted to ICU 11 ± 9 days after symptom onset. The most common symptoms on admission were cough (19 cases), fatigue or myalgia (18 cases), fever (17 cases) and dyspnea (16 cases). Eleven (55%) of the patients had underlying diseases, among which hypertension was the most common (11 cases), followed by cardiovascular disease (4 cases) and diabetes (3 cases). Six (30%) of the patients received invasive mechanical ventilation and continued renal replacement therapy but eventually died. Acute cardiac injury was the most common complication (19 cases). Half of the patients died between the 2nd and 19th day after ICU admission. Compared with dead patients, the surviving patients had a lower average body weight (61.70±2.36 68.60±7.15 kg, =0.01) and a higher Glasgow Coma Index (14.69 ± 0.70 12.70 ± 2.45, =0.03), and were less likely to develop shock (2 10, =0.001) or acute respiratory distress syndrome (2 10, =0.001).@*CONCLUSIONS@#Critically ill patients with COVID-19 are generally older. A higher body weight and a lower lymphocyte count are potentially associated with a greater likeliness of fatality in ICU patients with COVID-19.

Resveratrol ameliorates intestinal barrier injury in rats with hemorrhagic shock through superoxide dismutase 2 activation / 中国中西医结合急救杂志

Objective To explore the protective effect of resveratrol (RSV) on intestine barrier injury induced by hemorrhagic shock and its mechanism in rats.Methods According to random number table method,sixty-four SPF grade male Sprague-Dawley (SD) rats were divided into four groups:Sham operation group (only the catheters were indwelled in arterial and venous passages after anesthesia),hemorrhagic shock model group (model group,the catheters were indwelled in arterial and venous passages after anesthesia,and 0.3 mL solvent was administrated after hemorrhagic shock),RSV group (the catheters were indwelled in arterial and venous passages after anesthesia,15 mg/kg RSV was administered after hemorrhagic shock),superoxide dismutase 2 (SOD2) specific inhibitor,2-Methoxyoestradiol (2-ME) group (on the basic treatment of RSV group,0.1 mmol/L 2-ME was administered).The hemorrhagic shock model was reproduced by femoral artery bleeding.After drug administration,all rats were divided into two parts.One part was used for observations on 24-hour survival rate and survival time,while in the other part,2 hours after the hemorrhagic shock,the blood was collected for determination of the content of serum D-lactic acid,and afterward the rats were executed to obtain small intestine tissues for the examination of histopathological changes and Chiu's score.Moreover,differences of expression levels of tight junction proteins (Occludin,Claudin,ZO-1) of small intestine tissue and the oxidative stress related indexes SOD2 activity and reduced glutathione (GSH),oxidized glutathione (GSSH),malonaldehyde (MDA) contents were compared among the groups.Results Compared with the sham group,the model group demonstrated decreased survival rate,SOD2 activity,GSH content,GSH/GSSH ratio,reduced survival time,significantly increased serum D-lactic acid activity,Chiu's score and MDA content,and decreased expressions of tight junction proteins in small intestine tissue.Compared with model group,the RSV group showed significant increased survival rate [75.0％ (6/8) vs.37.5％ (3/8)] and prolonged survival time (hours:21.0±4.3 vs.10.4±5.8,P ＜ 0.05),significantly decreased serum D-lactic acid (μg/L:380.18 ± 70.59 vs.500.88 ± 97.53) and Chiu's score (1.75 ± 0.71 vs.4.00± 0.53) in small intestine (both P ＜ 0.05),obviously increased expressions of tight junction proteins,SOD2 activity,GSH and GSH/GSSG [Occludin (gray value):0.89 ± 0.10 vs.0.43 ± 0.77,Claudin (gray value):0.78±0.06 vs.0.33 ± 0.05,ZO-1 (gray value):0.83 ± 0.06 vs.0.34 ± 0.07,all P ＜ 0.05],and the elevated SOD2 activity (kU/L:0.85 ± 0.12 vs.0.51 ± 0.11,P ＜ 0.05],as well as increased GSH content and GSH/GSSG ratio [GSH (μmol/L):7.25±1.01 vs.3.86±0.54,GSH/GSSG:6.39± 1.14 vs.1.56±0.25,both P ＜ 0.05] in the small intestine,and markedly reduced MDA content (ng/g:5.00± 1.31 vs.8.63±0.92,P ＜ 0.05).Compared with RSV group,the 2-ME group demonstrated significantly decreased survival rate [50.0％ (4/8) vs.75.0％ (6/8)] and further shorter survival time (hours:12.2 ± 5.7 vs.21.0±4.3),increased serum D-lactic acid (μg/L:463.88 ± 60.16 vs.380.18 ± 70.59),obviously elevated Chiu's score (3.13 ± 0.99 vs.1.75±0.71,P ＜ 0.05),decreased expressions of tight junction proteins [Occludin (gray value):0.55±0.04 vs.0.89±0.10,Claudin (gray value):0.38±0.05 vs.0.78±0.06,ZO-1 (gray value):0.41±0.04 vs.0.83±0.06,all P ＜ 0.05];moreover,the activity of SOD2,GSH content,GSH/GSSG ratio were greatly reduced [SOD2 activity (kU/L):0.58 ± 0.13 vs.0.85 ± 0.12,GSH (μmol/L):4.49 ± 0.52 vs.7.25 ± 1.01,GSH/GSSG:1.57 ± 0.39 vs.6.39 ± 1.14,all P ＜ 0.05],and increased MDA content (ng/g:6.25 ± 1.04 vs.5.00 ± 1.31,P ＜ 0.05).The small intestine tissue was basically normal in Sham group,and no significant pathological changes were seen;in the model group,the small intestine epithelial mierovilli were collapsed and the mucosal barrier was destroyed obviously;in the RSV group the damages of small intestine microvilli and barrier were markedly alleviated;in 2-ME group the pathological changes were more evident compared with those in the RSV group.Conclusion RSV can improve intestinal barrier injury following hemorrhagic shock in rats;its mechanism may be related to SOD2 activation.

Diagnostic value of transcranial Doppler ultrasound in early stage cerebral arteriosclerosis / 中南大学学报(医学版)

OBJECTIVE@#To investigate the diagnostic value of transcranial Doppler ultrasound in early stage cerebral arteriosclerosis.@*METHODS@#We selected 50 patients with early cerebral arteriosclerosis as the disease group. At the same time we selected another 50 patients as a control group with no significant symptoms in the nervous system. By 2 MHz pulse Doppler probe through double-temporal windows and pillow windows Basilar artery (BA), the bilateral middle cerebral artery (MCA) was detected. In the TCD spectrum, we selected the spectrum of a single-family cardiac cycle, identified the starting point (ts), pulse-incisure point (ti), end point (te), and the peak velocity (tp), measured the time of the spectrum starting point to the peak velocity (Tp) and calculated the time required for the peak velocity in the share of ventricular systolic (Tp/Ti), the time required for the peak velocity in the share of the whole cardiac cycle (Tp/T). Tp, Tp/Ti and Tp/T were respectively named as time to peak velocity (TPV), peak-time index-1 (PTI-1) and peak-time index-2 (PTI-2). All data were analyzed by SPSS13.0.@*RESULTS@#There was no significant difference in blood vessel velocity, PI, RI and S/D of BA and RMCA (P>0.05) between the control group and the disease group. Compared with the control group, TPV of the BA, LMCA and RMCA significantly extended, PTI-1 and PTI-2 of BA, LMCA and RMCA increased significantly in the disease group (P0.05).@*CONCLUSION@#TPV, PTI-1 and PTI-2 are sensitive indicators of early stage cerebral arteriosclerosis.

Expressions of MDM2 and VEGF in osteosarcoma tissue and clinical significances / 吉林大学学报(医学版)

Objective To explore the relation of the expressions of MDM2 and VEGF in osteosarcoma with the pathological parameters and prognosis of the tumor.Methods The expressions of MDM2 and VEGF were detected with immunohistochemical(SP) method in specimens from 56 cases of osteosarcoma.The correlation between the expressions of MDM2,VEGF and pathological grade,metastasis and prognosis was analyzed statistically.while 8 cases of fibrous dysplasia of bone were used as negative control group.Results The positive rates of MDM2 and VEGF in osteosarcoma were 64.3%(36/56) and 67.9%(38/56),respectively .MDM2 and VEGF didn't express in negative control group.The expression of MDM2 and VEGF were not significantly correlated to the pathological grades of the osteosarcoma,but which were significantly correlated with tumor metastasis and prognosis(P