Vampire bat saliva contains a plasminogen activator that presumably assists these hematophagous animals during feeding. Bat-PA (H), the full-length form of Vampire Bat Salivary Plasminogen Activator (DSPAalpha1), is homologous and similar efficacy to tissue-type plasminogen activator (t-PA). The strict fibrin dependence of activity is a characteristic which could be desirable in the fibrinolytic therapy. It is a unique fibrinolytic enzyme that does not promote neurodegeneration. In this study, according to the reported gene sequence (GenBank Accession No. J05082) of Vampire bat (D. rotundus) plasminogen activator. It was the first time to synthesize the full sequence of DSPAalpha1 in vitro and clone it into the expression vector pPIC9K, the recombinant plasmid was linearized and transformed into Pichia pastoris GS115 strain. Secreted expression of recombinant DSPAalpha1 was attained by methanol induction and its molecular mass is 47 kD. To get recombinant GS115 with high amount of protein, hundreds of His+ transformants had been screened to isolate clones resistant to high levels G418 (2-4 mg/mL), the selected clones mini-expressed in Pichia pastoris, and tested their fibrinolytic activities and expressed protein bands by fibrin plate assay and SDS-PAGE. DSPAalpha1 was determined by optical density after SDS-PAGE, the yield is about 30 mg per liter of fermentation culture. DSPAalpha1 derived often from mammalian cells: Chinese hamster ovary (CHO) cells, Baby hamster kidney (BHK) cells, COS cells, which might be produced at high cost. In Pichia pastoris, it is expected to higher yield and lower cost, thus it might be able to serve as new thrombolytic candidate.
Animals ; Base Sequence ; Chiroptera ; genetics ; Genetic Vectors ; genetics ; Molecular Sequence Data ; Pichia ; genetics ; metabolism ; Plasminogen Activators ; biosynthesis ; genetics ; Recombinant Proteins ; biosynthesis ; genetics ; Recombination, Genetic ; Sequence Analysis, DNA

Objective To investigate the influencing factors of short-term and long-term outcomes in patients with acute ischemic stroke. Methods From March 2015 to March 2016, patients with acute ischemic stroke treated at the Department of Neurology, the Second Hospital of Tianjin Medical University were enrolled prospectively. The demographic and baseline clinical data of the patients were recorded. The modified Rankin scale(mRS)was used to assess the short-term outcomes at 14 d and one year after onset. Good outcome was defined as mRS 0-2, and poor outcome was defined as > 2. Death or new cardiocerebrovascular events were recorded.Multivariable logistic regression analysis was used to determine the influencing factors of short-term and long-term outcomes in patients with acute ischemic stroke.Results A total of 514 patients with acute ischemic stroke were enrolled, including 338 (65.8％) with short-term good outcome and 176 (34.2％) with poor outcome. There were significant differences in age, atrial fibrillation, history of past stroke or transient ischemic attack (TIA), etiological typing of stroke, baseline NIHSS score, severity of stroke, pulmonary infection, dysphagia, and urinary incontinence between the short-term good outcome group and the poor outcome group (P<0.05). Multivariable logistic regression analysis showed that history of past stroke or TIA(odds ratio[OR]2.188, 95％ confidence interval[CI] 1.192-4.014;P=0.011),baseline NIHSS score(OR 1.504,95％ CI 1.362-1.661;P=0.001),and urinary incontinence (OR 4.114, 95％ CI 1.934-8.751; P=0.001) were the independent influencing factors of short-term outcome. A total of 467 patients completed 1-year follow-up, including 315 (67.5％)with long-term good outcome and 152(32.5％)with poor outcome.There were significant differences in age,history of ischemic heart disease, stroke or TIA, etiological typing of stroke, baseline NIHSS score, severity of stroke, pulmonary infection, dysphagia, urinary incontinence, secondary prevention of antiplatelet drugs and statins between the long-term good outcome group and the poor outcome group(all P<0.05).Multivariable logistic regression analysis showed that age(OR 1.029,95％ CI 1.004-1.055; P=0.022), past history of stroke or TIA(OR 1.983, 95％ CI 1.082-3.633; P=0.027), baseline NIHSS score (OR 1.271, 95％ CI 1.153-1.400; P=0.001), urinary incontinence (OR 4.996, 95％ CI 2.308-10.813; P= 0.001), and secondary prevention using antiplatelet drugs (OR 0.227, 95％ CI 0.125-0.414; P=0.001) were the independent influencing factors of long-term outcome. The baseline NIHSS score (OR 1.184, 95％ CI 1.070-1.310;P<0.001) and secondary prevention using antiplatelet agents (OR 0.064, 95％ CI 0.014-0.284; P< 0.001) were the independent influencing factors of death or occurring vascular events. Conclusion Age, past history of stroke or TIA, baseline NIHSS score, urinary incontinence, antiplatelet drugs for secondary prevention were independently associated with the outcomes in patients with acute ischemic stroke. Early and targeted intervention of modifiable factors and the emphasis on the use of antiplatelet agents in secondary prevention might reduce stroke recurrence and improve outcomes.

Objective:To investigate the clinicopathological features, immunophenotype and differential diagnosis of clear cell hidradenoma, and to analyze the origin of clear cell hidradenoma and the underlying mechanism.Methods:The clinical data of 23 cases of clear cell hidradenoma who underwent surgical resection in Suzhou Municipal Hospital between December 2017 and July 2021 were retrospectively analyzed. Clinical manifestation, imaging features, pathological features and prognosis of the 23 cases of clear cell hidradenoma were analyzed. Expression levels of epithelial membrane antigen, cytokeratin 20, cytokeratin 7, cytokeratin 14, carcinoembryonic antigen, and gross cystic disease fluid protein 15 were detected by immunohistochemical staining technique using the EnVision system. Periodic acid-Schiff (PAS) staining was performed to visualize glycogen.Results:Among the 23 cases, 8 were male and 14 were female, aged 14-94 years, with a median age of 55 years. The first symptom of clear cell hidradenoma was epidermal bulgels in 18 cases.Contrast ultrasonography showed a subcutaneous cystic solid echo mass with abundant blood flow in the solid part. The tumor histologically consisted of two types of cells: secretory epithelial cells or glandular epithelial cells and clear cells. Twenty cases had tumors with the features of benign clear cell hidradenoma. Two cases had atypical clear cell hidradenoma with atypia and mitosis. One case had malignant clear cell hidradenoma. Tumor cells were positive for epithelial membrane antigen, cytokeratin 7, cytokeratin 14, carcinoembryonic antigen, and gross cystic disease fluid protein 15 and they were Periodic acid-Schiff-positive. Twenty-three patients were followed up for 2-36 months, of which 4 were lost to follow-up and the rest had no recurrence of clear cell hidradenoma.Conclusion:Clear cell hidradenoma is rare and has a good prognosis. Malignant clear cell hidradenoma is rarer and has a poor prognosis. Diagnosis of clear cell hidradenoma is mainly based on comprehensive analysis of pathological features and immunophenotypes. Clear cell hidradenoma should be differentiated from metastatic clear cell carcinoma, spiral adenoma, cortical adenoma, and malignant melanoma.

Objective:To investigate urinary microalbumin to creatinine ratio (ACR) and α1-microglobulin to creatinine ratio (MCR) of people aged 40 years old and above in Shanxi province, and analyze the influencing factors of abnormal ACR and MCR, and to provide evidence for the prevention and control of chronic kidney diseases.Methods:It was a cross-sectional study. The data came from a screening study of chronic kidney diseases conducted by Shanxi Provincial People's Hospital from April to November 2019, involving aged 40 years old and above from 10 counties (Ningwu county, Yu county, Yangqu county, Lin county, Shouyang county, Zezhou county, Huozhou city, Hejin city, Linyi county and Ruicheng county) in Shanxi province. The related data were collected through questionnaire surveys, physical examinations, and blood and urine sample collection. Urinary α1-microglobulin, creatinine, and microalbuminuria were measured. Urinary ACR and MCR were calculated using urinary creatinine correction. ACR abnormality was defined as ≥30 mg/g, and MCR abnormality was defined as >23 mg/g. Covariate analysis was used to control confounding factors, and adjusted urinary ACR and MCR of 10 counties were calculated. Spearman correlation analysis and chi-square test were performed to analyze the factors associated with abnormal urinary ACR and MCR. Logistic regression analysis model was used to identify the influencing factors of abnormal urinary ACR and MCR.Results:A total of 12 285 residents were enrolled in the study, including 5 206 males (42.4%) and 7 079 females (57.6%). The median age was 58.0 (51.0, 66.0) years old. The median urinary ACR was 7.5 (4.5, 15.7) mg/g, and the median urinary MCR was 10.2 (6.4, 16.2) mg/g. A total of 1 572 individuals (12.80%) had urinary ACR abnormality and 1 450 individuals (11.80%) had urinary MCR abnormality. Yangqu county, Yuxian county, and Ningwu county had higher urinary ACR with (35.58±3.04) mg/g, (34.08±4.50) mg/g and (32.09±3.19) mg/g, respectively. The urinary MCR was generally similar among the 10 counties and Yangqu county had higher urinary MCR with (13.86±0.41) mg/g. In addition to Yu county, female individuals had higher urinary ACR compared to males in other counties, whereas female individuals had lower urinary MCR compared to males in 10 counties. Multivariate logistic regression analysis results showed that elevated triglyceride, fasting blood glucose, glycated hemoglobin, systolic blood pressure, diastolic blood pressure, age, body mass index and gender were independent influencing factors of abnormal urinary ACR and MCR (all P<0.05). Elevated blood homocysteine and low educational level were independent influencing factors of urinary MCR abnormality (both P<0.05). Conclusions:There are differences of gender and region in urinary ACR and MCR among individuals aged 40 years old and above in the 10 counties of Shanxi province. Triglyceride, fasting blood glucose, glycated hemoglobin, systolic blood pressure, diastolic blood pressure, age, gender, and body mass index are independent related factors of abnormal urinary ACR and MCR. Blood homocysteine and education level are independent related factors of abnormal urinary MCR.

Sjögren's syndrome is a kind of autoimmune disease, whose main clinical symptoms are dry mouth, dry eye and chronic parotid glandular inflammation. The conservative treatments include artificial tears or saliva,oral administration of corticosteroids,and immunosuppressantsl with limited effectiveness. Along with the development of molecular biology, vast attentions are being paid to researches on gene therapy for Sjögren's syndrome, hopefully to bring gospel to patients with Sjögren's syndrome. This article reviews the recent research progresses on transcatheter delivery of recombinant adenovirus vector with aquaporin gene in experimental treatment of Sjögren's syndrome.
Adenoviridae ; Aquaporins ; genetics ; Autoimmune Diseases ; therapy ; Catheters ; Genetic Therapy ; Genetic Vectors ; administration & dosage ; Humans ; Sjogren's Syndrome ; therapy

Objective: To systematically review available risk prediction models evidence on construction and verification of colorectal cancer risk prediction models. Methods: "Colorectal neoplasms", "risk assessment", "colorectal cancer", "colorectal tumor", "colon cancer", "colon tumor", "rectal cancer", "rectal tumor", "anal cancer", "anal tumor", "risk prediction", "malignancy", "carcinogenesis", "model" were used as search keywords. Journal papers and grey literature were searched from Chinese electronic databases (CNKI and Wanfang) and English electronic databases (PubMed and Embase) from their inception to 30 Apr 2018. The language of literature was restricted to Chinese and English. The inclusion criteria were human-oriented researches with complete information for model construction,verification and evaluation. The exclusion criteria were informal publications such as conference abstracts, Chinese disertation papers, and non-primary research materials such as reviews,letters,and news reports. Descriptive characteristics,targeted population, study design, model construction method and prediction results were extracted. A total of 36 papers involving 27 models were included. The population characteristics of all included studies,the type of research, the method of model construction and the prediction results of the model were analyzed. Results: As for model construction,there were 13 European and American population based model studies,14 Asian population based model studies,including 7 Chinese mainland based model studies. According to the factors selected into the model, these models can be divided into traditional epidemiological models (17 models), clinical index combined models (4 models),and genetic susceptibility index combined models (6 models). As for model verification,only 9 models were cross-verified in the internal population after model construction, and the extrapolation of model prediction effect was not effectively evaluated; 17 models were verified in an external population; there was only one model verified in two external populations in terms of risk prediction effect; the area under the curve of 27 models was 0.56-0.85. Conclusion The risk prediction model of colorectal cancer is in the development stage. The external evaluation of model prediction effect is less and the prediction ability is not good, and the existing models have limited exploration of clinical indicators.

Purpose To explore the value of insulinoma associated protein 1(INSM1)in the pathological diagnosis and differential diagnosis of medullary thyroid carcinoma(MTC).Methods Twenty-eight cases of MTC were collected as the ex-perimental group and 25 cases of thyroid tumors(6 cases of transparent trabecular tumors,16 cases of poorly differentiated cancer,2 cases of spindle cell papillary carcinoma,and 1 case of thyroid paraganglioma)that needed to be differentiated from MTC in the same period were collected as the control group.The expression of INSM1 was detected using immunohistochemical EnVision method,and the sensitivity and specificity of INSM1 in MTC were analyzed,and compared with traditional MTC markers(Syn,CgA,CT,and CEA).Results The positive rate of IN-SM1 in the MTC group was96.4%(27/28),significantly high-er than the control group of 4.0%(1/25)(P<0.05).INSM1 had a sensitivity of 96.4%in MTC,significantly higher than CgA(71.4%)(P<0.05),and was positively expressed in CT negative MTC cases.The specificity of INSM1 in MTC was 96.0%,significantly higher than that of CEA(72.0%)(P<0.05).Conclusion INSM1 has high sensitivity and specificity in MTC,with a nuclear positive staining pattern,clear back-ground,and easy identification.It is superior to the traditional biomarkers and has high diagnostic value in MTC.

Objective The objective of this study was to explore the association between human papillomavirus( HPV) and prognosis of lung cancer by meta-analysis. Methods The PubMed,Embase and Cochrane literature databases studies were searched using a combination of subject terms and free words. As of October 2018,a total of 123 related documents were obtained. After screen-ing the literature according to inclusion and exclusion criteria,the basic information of the study,HPV detection methods,lung cancer patients,hazard ratio(HR)values and 95% confidence interval(CI)were extracted from each study. The meta-analysis of random effects models was used to evaluate the correlation between HPV infection and prognosis in patients with lung cancer. Heterogeneity was assessed using the Q test and I2statistics,and publication bias was tested using Egger′s linear regression test and Begg′s rank cor-relation test. Results The study finally included 11 articles(9 in Asia,2 in Europe and US),and 1439 patients with lung cancer. Meta-analysis using a random-effects model showed no significant association between HPV infection and prognosis of lung cancer (HR=0. 90,95% CI:0. 71~1. 13). A stratified analysis of lung cancer pathological subtypes showed that the prognosis of patients with HPV-infected lung adenocarcinoma was significantly better than that in patients without HPV-infected lung adenocarcinoma (HR=0. 65,95% CI:0. 49~0. 85). Sensitivity analysis was performed by sequentially removing the included studies,and the results were not statistically significant. The results of Egger′s test(P=0. 708)and Begg′s test(P=0. 784)suggest that there is no publica-tion bias in this study. Conclusion HPV infection may be related to the prognostic of patients with lung adenocarcinoma. More basic and clinical studies are needed to further explore the association between HPV infection and lung adenocarcinoma as well as the corre-sponding mechanisms in the future.

ObjectiveTo analyze the coping style and its relationship with anxiety status among middle school students aged 13 to 18 in Anhui province during the COVID-19 epidemic. MethodsFrom February 13 to 19， 2020， a cross-sectional survey was conducted among middle school students in Anhui province by using convenience sampling method and network questionnaire. The assessment tools included the Coping Style Scale for Middle School Students （CSSMSS） and the Screen for Child Anxiety Related Emotional Disorders （SCARED）. ResultsThe results of the CSSMSS evaluation showed that female students scored higher in emotion abreaction than male students ［（8.27±2.98） vs. （7.84±2.91）］. The scores of tolerance ［（9.74±2.73） vs. （9.11±2.60）］， escape ［（7.82±2.79） vs. （7.26±2.44）］， emotion abreaction ［（8.48±2.97） vs. （7.91±2.93）］ and fantasy/denial ［（9.79±3.56） vs. （9.26±3.47）］ of senior high school students were higher than those of junior high school students， and the score of problem solving ［（19.38±4.07） vs. （20.33±4.54）］ was lower than that of junior high school students， with statistical significance （P<0.05 or 0.01）. Correlation analysis results showed that the scores of tolerance， escape， emotion abreaction and fantasy/deny in CSSMSS were positively correlated with the scores of SCARED of middle school students in Anhui province （r=0.348， 0.287， 0.390， 0.501， P<0.01）. ConclusionDuring COVID-19 epidemic， students of different genders and grades in Anhui province have different coping styles， and some coping styles may induce anxiety status.

Animals ; Cell Differentiation ; Hepatocyte Growth Factor ; genetics ; metabolism ; Membrane Proteins ; genetics ; metabolism ; Mice ; Mouse Embryonic Stem Cells ; cytology ; metabolism ; Protein Precursors ; genetics ; metabolism ; Serine Endopeptidases ; genetics ; metabolism