[Clinical implications of thrombolytic therapy for patients with acute intermediate-high risk pulmonary embolism in Tibet plateau, China].

At present, clinical studies and case reports of systemic thrombolytic therapy for patients with acute pulmonary embolism in Tibet Plateau are very rare. There is little understanding of the risk factors, clinical characteristics, and thrombolytic therapy for patients with acute pulmonary embolism at intermediate-high risk in Tibet Plateau. In this paper, we reported the data of 4 patients with acute intermediate-high risk pulmonary embolism treated with thrombolytic therapy in Lhasa People's Hospital. The demographic characteristics, clinical manifestations, treatment, and outcomes were analyzed. We summarized the clinical features and raised scientific issues. We aimed to provide basic data to improve the standardized diagnosis and treatment of acute pulmonary embolism in plateau, and to point out the direction of future clinical research in this field.

[Clinical observation of superior rectus transposition with/without augmented suture and vertical rectus transposition for the treatment of strabismus caused by complete abducens nerve palsy].

Objective: To evaluate the efficacy and safety of superior rectus transposition (SRT) with/without augmented suture and vertical rectus transposition (VRT) for the treatment of strabismus caused by complete abducens nerve palsy. Methods: This was a retrospective cohort study. Forty-two patients (42 eyes) with complete abducens nerve palsy underwent strabismic surgeries from January 2015 to November 2020 in Tianjin Eye Hospital. According to the different procedures, the patients were divided into three groups: SRT group (16 cases, SRT with medial rectus recession), superior rectus transposition with augmented suture (SRTA) group (13 cases, SRT with Buckley suture and medial rectus recession) and VRT group (13 cases). The preoperative and postoperative (1, 6 and 12 months) data including deviations, ocular motility, binocular vision and surgical complications among three groups were analyzed and compared. χ2 test was used for comparison of count data among three groups. The measurement data were compared among three groups by the repeated measures ANOVA. LSD-t test was used for within-group comparison and between-group comparison. Results: There was no difference in sex ratio, age and course of disease among the groups (all P>0.05). The horizontal deviations of the three groups at 1, 6 and 12 months after surgeries was lower than that before surgeries, and the difference was statistically significant (all P<0.001). The horizontal deviations of the SRT group, SRTA group and VRT group at 12 months after surgeries were (+0.8±5.8), (+0.8±4.5), (+1.2±2.5) prism diopters (PD), respectively, lower than that of the preoperative (+82.8±17.2), (+77.7±26.1), (+71.5±18.6) PD. However, there was no significant difference among different postoperative follow-up timepoints (all P>0.05). There was no difference in horizontal deviations before surgeries and at 1, 6 and 12 months after surgeries among three groups (P>0.05). There were significant differences in the scales of abduction motility among preoperative, postoperative 1, 6 and 12 months measurements for three group (all P<0.001). The scales of abduction before surgeries in the SRT group, SRTA group, and VRT group were (-4.4±0.5), (-4.4±0.5), (-4.5±0.5) scale and at 12 months after surgeries were (-2.3±0.7), (-2.2±0.5), (-2.1±0.6) scale respectively. But there was no change among different postoperative follow-up timepoints (all P>0.05). Preoperative and postoperative 1-, 6-and 12-month abduction motility was similar among three groups (P>0.05). There were significant differences in the scales of adduction limitation among preoperative, postoperative 1-, 6-and 12-months measurements for three group (all P<0.05). But there was no change among different postoperative follow-up timepoints (all P>0.05). There were significant differences between the SRT group [(-0.9±0.6), (-0.8±0.6) scale] and the SRTA groups [(-1.5±0.5), (-1.4±0.5) scale] (t=-2.62, -2.52) and between the SRTA group and the VRT group [(-0.8±0.8), (-0.6±0.7) scale] (t=2.62, 3.01) at 6 and 12 months after surgeries (all P<0.05). The outcomes of binocular vision at postoperative 12 months were similar among three groups (P>0.05). No patient had torsional diplopia and anterior segment ischemia. Only 2 patients from the SRTA group had hypotropia of 4 to 5 PD in the primary position associated with supraduction limitation. Conclusions: SRT with/without augmented suture and VRT are effective and safe procedures for the treatment of strabismus caused by complete abducens nerve palsy. They could correct deviations, improve abduction motility and restore binocular vision, with stable outcomes and a small risk of vertical and torsional diplopia.

[Multiple pulmonary nodules with interstitial changes].

A 53-year-old female patient with pulmonary nodules for more than 3 years was admitted to Beijing Chao-Yang Hospital because of cough and sputum with shortness of breath after exercise for 4 months. In the first two and a half years, her pulmonary nodules remained stable, after that the nodules increased obviously with interstitial changes. After admission, a venous thromboembolic (VTE) event was quickly detected with a marked increase in D-dimer. Then, based on the clues of VTE examination, bronchoscopy, gastroscope, positron emission tomography-CT, head magnetic resonance and other examinations were performed. The final pathological diagnosis was lung adenocarcinoma, mainly solid with mucus secretion, with mediastinal hilar lymph node metastasis, intrapulmonary metastasis and gastric metastasis. Gene detection of lung and stomach histopathological tissues showed positive EML4-ALK fusion gene. The patient received therapies with crizotinib, alectinib in sequence and anticoagulation. After 20-month treatment, a telephone follow-up showed that there was no significant limitation in her daily activities.

[Advances in genome-wide association study of pulmonary arterial hypertension].

Pulmonary arterial hypertension (PAH) is a complex disease caused by multiple factors, including idiopathic PAH, heritable PAH, disease related PAH etc. Due to the high genetic heterogeneity, clinical characteristics and prognosis of PAH patients vary greatly. At present, the specific pathogenesis of PAH is unclear, and the diagnosis and treatment of PAH remain to be explored. Therefore, the study of genetic susceptibility to PAH is of great significance for understanding the occurrence and development of the disease. With the development of genome-wide association study (GWAS), a large number of genetic variations related to etiology, clinical manifestations, prognosis and treatment of PAH have been identified. This review summarizes the recent progress in the application of GWAS in the study of genetic susceptibility of PAH, and provides new insights for further exploration of the development and individualized management of PAH.

[Research progress of non-invasive examination in the screening of chronic liver disease among the general community population].

Due to its insidious nature, chronic liver disease usually only appears after a decompensating event, and delays in diagnosis lead to a high mortality. Non-invasive examination can help diagnose and intervene early stage liver fibrosis in patients with chronic liver disease. In a hospital population, this test has been widely used and appropriate; however, its value is unclear in the general community population. In recent years, there have been many studies on non-invasive examinations for foreign community populations. These non-invasive examination tools can screen patients with chronic liver disease and liver cirrhosis among the general community population. Further studies have found that screening community populations with liver disease risk factors (heavy alcohol consumption, type 2 diabetes mellitus, and obesity) can more efficiently screen out patients with chronic liver disease in the community population. In brief, these noninvasive examination can find a large number of previously undiscovered liver disease patients in the community population, compared to traditional community screening that rely on abnormal liver function testing. This paper discusses the non-invasive examination in the screening of chronic liver disease among the community population, so as to find a pioneering community non-invasive liver disease screening path for our country.

VASPIN reduces inflammation and endoplasmic reticulum stress of renal tubular epithelial cells by inhibiting HMGB1 and relieves renal ischemia-reperfusion injury.

OBJECTIVE: Renal ischemia-reperfusion injury (IRI) is a clinically common issue and the resulting acute kidney injury (AKI) seriously threatens the patient's life. Therefore, prevention and treatment of renal IRI are the key to alleviating AKI in such patients. The purpose of this study was to explore the effects of VASPIN on mouse renal IRI and human renal proximal tubular epithelial cells (HK-2 cells) to provide a new direction for the treatment of clinical renal IRI. MATERIALS AND METHODS: C57/BL6 mice were used to construct a renal IRI model and recombinant mouse VASPIN was subcutaneously injected to determine whether VASPIN can alleviate renal IRI in mice by histological examination and detection of mouse urine and serum related indicators. In addition, HK-2 cells were cultured and an IRI model was constructed at the cellular level by hypoxia reoxygenation to examine the effect and mechanism of VASPIN on endoplasmic reticulum stress (ERS) in HK-2 cells. RESULTS: Results revealed that in VASPIN-treated mice, edema of renal tubular epithelial cells was significantly improved and renal injury markers netrin-1 and L-FAPB were decreased in urine. In addition, VASPIN also reduced the expression of inflammatory factors in mouse serum and the level of oxidative stress in kidney tissue. The expression of ERS-related molecules (GRP78, ATF6, caspase12, and CHOP) in HK-2 cells treated with VASPIN was significantly reduced and VASPIN decreased the expression of the pro-inflammatory factor HMGB1. Moreover, VASPIN promoted the activity of the Nrf2/ARE/HO-1 signaling pathway and inhibited the NF-кB signaling pathway by inhibiting HMGB1. CONCLUSIONS: VASPIN reduces inflammation and ERS levels in kidney tissue and attenuates renal IRI by activating the Nrf2/ARE/HO-1 signaling pathway and inhibiting the NF-кB signaling pathway via inhibition of HMGB1.

[Application value of non-invasive ventilation combined with high flow nasal cannula oxygen therapy in sequential treatment of patients with chronic obstructive pulmonary disease after mechanical ventilation].

Objective: To investigate the value of non-invasive ventilation (NIV) combined with high flow nasal cannula oxygen therapy (HFNCO) in sequential treatment of patients with chronic obstructive pulmonary disease after mechanical ventilation. Methods: Chronic obstructive pulmonary disease with acute exacerbation (AECOPD) patients with invasive mechanical ventilation (MV) and successful withdrawal admitted into Huxi Affiliated Hospital of Jining Medical College from January 2018 to December 2019 were enrolled for perspective study. The patients were divided into treatment group (n=40) and control group (n=33) by random number table method. The treatment group was given NIV and HFNCO, the control group was given NIV treatment alone. Bedside ultrasound was used to measure the patients' diaphragmatic motion, and the differences between the two groups of patients before treatment, 24, 48 and 72 h after treatment were compared in diaphragmatic excursions during quiet breathing (DEq), diaphragmatic excursions during deep breathing(DEd), diaphragmatic shallow fast breathing index (D-RSBI), arterial oxygen partial pressure (PaO(2)), arterial partial pressure of carbon dioxide (PaCO(2)), re-tracheal intubation rate, mortality rate for 28 days and average duration of NPPV treatment within 3 days. Results: There were no statistically significant differences in DEq, DEd, D-RSBI, PaO(2) and PaCO(2) between the two groups before treatment (all P>0.05). After 24 h treatment, DEd decreased in both groups, D-RSBI increased in both groups, However, D-RSBI [(1.33±0.56) vs (1.62±0.59) times·min(-1)·mm(-1)] in the treatment group was significantly lower than the control group, P=0.034. After 72 h treatment, DEd [(41.4±8.1) vs (37.8±6.0) mm] was significantly higher than the control group, D-RSBI [(1.02±0.27) vs (1.22±0.43) times·min(-1)·mm(-1)] was significantly lower than the control group (all P<0.05). The average duration of NIV treatment time [(7.5±1.2) vs (9.3±2.6) h] in the treatment group was significantly shorter than that in the control group (P<0.01). There were no statistically significant differences in PaO(2), PCO(2), re-tracheal intubation rate and the mortality rate of 28 days. Conclusion: NIV combined with HFNCO sequential therapy can effectively relieve diaphragm fatigue and promote recovery of respiratory muscle strength, and it's better than NIV alone.

[The surgical effect of secondary esotropia with diplopia after orbital decompression for thyroid-associated ophthalmopathy].

Objective: To evaluate the effect of extraocular muscle surgery in treating secondary esotropia with diplopia after orbital decompression for thyroid-associated ophthalmopathy (TAO). Methods: Retrospective case series study. Eleven secondary esotropic patients with diplopia after orbital decompression for TAO who underwent extraocular muscle surgery during March 2016 and October 2018 in Tianjin Eye Hospital were included. All patients had new onset esotropia after decompression surgery and underwent strabismus surgery. Deviation angle, diplopia, and ocular movement were observed preoperatively and postoperatively. Surgeries were carried out under the monitored anesthesia care combined with local anesthesia, and the eye alignment was adjusted to orthotropia with no diplopia in the operation with the technique of intraoperative adjustable suture and forced duction test results. All the patients were followed up. Results: The ages of the patients were from 26 to 42 years (1 male and 10 females). The deviation angles were 10-98 prism diopter. Orbital CT scan showed that the horizontal rectus thickness was increased at different levels, and the thickness of the medial rectus was increased more than that of the lateral rectus. Two patients had monocular medial rectus recession only, 2 had bilateral medial rectus recession only, 2 had monocular medial rectus recession combined with lateral rectus resection, and the other 5 had bilateral medial rectus recession with monocular lateral rectus resection. The recession of medial rectus muscle was 3.5 to 7.5 mm in 11 patients and 2.0 to 6.0 mm in 7 patients with the resection of lateral rectus muscle. Diplopia of the 11 patients disappeared with orthotropia at primary gaze position after the extraocular muscle surgery. The limitation of abduction changed from preoperative (-1.91±1.04) to postoperative (-0.64±0.81). All the patients were satisfied with the surgical results. At the end of the follow-up (6 to 24 months), the results of the patients were stable, and no over-correction was found. Conclusion: Extraocular muscle surgery with intraoperative adjustable suture technique can effectively treat TAO patients with secondary esotropia with diplopia after orbital decompression. (Chin J Ophthalmol, 2020, 56: 183-188).

[Specifications for diagnosis and treatment of non-neonatal tetanus].

Tetanus consists of neonatal tetanus and non-neonatal tetanus. Non-neonatal tetanus remains a serious public health problem, although neonatal tetanus has been eliminated in China since 2012. Non-neonatal tetanus is a potential fatal disease. In the absence of medical intervention, the mortality rate of severe cases is almost 100%. Even with vigorous treatment, the mortality rate is still 30%-50% globally. These specifications aim to regulate non-neonatal tetanus diagnosis and treatment in China, in order to improve medical quality and safety. These specifications introduce the etiology, epidemiology, pathogenesis, clinical manifestations and laboratory tests, diagnosis, differential diagnosis, grading and treatment of non-neonatal tetanus.

[Specifications for diagnosis and treatment of non-neonatal tetanus].

Tetanus consists of neonatal tetanus and non-neonatal tetanus. Although neonatal tetanus in China has been eliminated since 2012, non-neonatal tetanus remains a serious public health problem. Non-neonatal tetanus is a potential fatal disease, and the mortality rate of severe cases is almost 100% in the absence of medical intervention. Even with vigorous treatment, the mortality rate is still 30~50% globally. In order to standardize the diagnosis and treatment of non-neonatal tetanus in China, this specification is hereby formulated. This standard includes etiology, epidemiology, pathogenesis, clinical manifestations, laboratory tests, diagnosis, differential diagnosis, classification, grading and treatment of non-neonatal tetanus.

[Analysis of clinical features and targeted drug therapy of portopulmonary hypertension].

Objective: To analyze the clinical characteristics and the effect of targeted drug therapy of portopulmonary hypertension (PoPH). Methods: A total of 5 patients with PoPH who were admitted to the Department of Pulmonary and Critical Care Medicine of Beijing Chao-Yang Hospital from January 1, 2017 to December 31, 2018 were included. The clinical information and follow-up data were collected. The patient's medical history, clinical manifestations, right cardiac catheterization (RHC), classification of cardiac and hepatic function, treatment and prognosis were analyzed. Results: Among the 5 patients with PoPH, 3 were male and 2 were female. The median age was 56 years. The underlying diseases of portal hypertension were all cirrhosis, and 1 patient combined with hepatopulmonary syndrome (HPS). Dyspnea was the main respiratory symptom in all the 5 patients, and the median time from symptom onset to diagnosis was 1 year (5 months to 8 years). RHC was used as the diagnostic criteria for pulmonary hypertension in all patients, with a median mean pulmonary arterial pressure of 42 mmHg (1 mmHg=0.133 kPa) and a median pulmonary vascular resistance of 538 dyn·s·cm(-5). 3 cases were in Child-Pugh liver function grade B, and 2 were in grade A. The hepatic reserve function was not matched with the severity of cardiac insufficiency. Liver transplantation was performed in 1 patient, whose right ventricular dysfunction can be alleviated by targeted drug therapy after operation. All the 5 patients received targeted drug therapy of pulmonary hypertension. In the 3 patients who were regularly treated with targeted drugs and followed up on time, the cardiac function was improved during the follow-up period. There was no improvement or even deterioration of cardiac function in 2 patients who were not regularly treated or followed up. One patient died after liver transplantation. The cause of death was severe pneumonia and right ventricular dysfunction. The survival time after transplantation was 1 year. Conclusions: In PoPH patients, the hepatic reserve function is not matched with the heart function classification. PoPH can coexist with HPS. Regular application of pulmonary hypertension targeting drugs may benefit patients with PoPH.

[Pulmonary cavities with Takayasu arteritis: report of 3 cases and literature review].

Objective: To analyze the clinical features of 3 cases of Takayasu arteritis(TA) with pulmonary cavities on chest computed tomography(CT). Methods: The clinical data of 3 TA patients with cavities on the chest CT who were admitted into Beijing Chaoyang Hospital were retrospectively analyzed. A literature search was performed with "Takayasu arteritis" and "pulmonary" as the key words in China Knowledge Resource Intergrated Database (CNKI) and Pubmed Database for publications from Jan 1, 2000 to Dec. 31,2017. The relevant literatures were reviewed. Results: Among the 3 patients, 2 were males and 1 was female, aging 49, 28 and 28 years, respectively. They presented with cough, fever and chest pain, and chest CT showed cavities, single or multiple, either with thick or thin wall, or wedge-shaped consolidation, residual stripes after being absorbed, and one case had pulmonary biopsy results which showed hemorrhagic infarction. They were all misdiagnosed before as pneumonia, pulmonary tuberculosis, pulmonary thromboembolism. After being treated by combination therapy of glucocorticoids and immunosuppressive agents, the disease improved significantly. A total of 777 cases with TA involving pulmonary arteries were reported, from which 13 cases with involvement of pulmonary parenchyma were described. Therefore total 16 cases including the 3 cases in this article were included for analysis. Twelve cases showed patchy or wedge-shaped ground-glass opacity and consolidation, and peripheral lung stripes remained after being absorbed. Two cases showed pleural effusion, and 4 cases showed cavities, 3 cases were misdiagnosed as pulmonary tuberculosis, 7 as pulmonary infection, and 5 as pulmonary thromboembolism. Conclusions: TA with pulmonary arteries involved is susceptible to be misdiagnosed and missed, and therefore, in patients with cough, hemoptysis, chest pain and cavities in pulmonary parenchyma, TA should be suspected. Early diagnosis and appropriate treatment can lead to a better prognosis.

Evaluation of the application of high volume hemofiltration in sepsis combined with acute kidney injury.

OBJECTIVE: To investigate the safety and effectiveness of the application of high-volume hemofiltration (HVHF) in children with sepsis combined with acute kidney injury. PATIENTS AND METHODS: A total of 76 child patients were enrolled and randomly divided equally (n=38) into control and the observation groups respectively. The control group received conventional volume hemofiltration (the ultrafiltration rate of 35-50 ml/kg/h), and the observation group received HVHF (50-100 ml/kg/h). RESULTS: The serum Interleukin-6 (IL-6), Tumor Necrosis Factor-a (TNF-α) and creatinine levels were significantly lower in the observation group than the control group at 6 h, 12 h, 24 h and 48 h of hemofiltration (p<0.05). The efficacy rate of treatment was improved. The mortality rate and incidence rate of complications were decreased, and the treatment course was significantly shortened (p<0.05). CONCLUSIONS: The application of HVHF in children with sepsis combined with acute kidney injury has a better safety and effectiveness.

Purification and characterization of a novel α-glucosidase from Malbranchea cinnamomea.

OBJECTIVES: To characterize a novel α-glucosidase from the thermophilic fungus Malbranchea cinnamomea. RESULTS: The enzyme was purified to homogeneity with purification fold of 40 and a recovery of 7.2 %. It was a monomer with molecular mass of 65.7 kDa on SDS-PAGE. It was optimally active at pH 6 and 50 °C (measured over 10 min) and exhibited a wide range of substrate specificity with the highest specific activity of 47.4 U mg(-1) for p-nitrophenyl α-D-glucopyranoside (pNPGlu) followed by isomaltose, panose and sucrose, suggesting that the enzyme belongs to the type I α-glucosidases. The K m values of the α-glucosidase for pNPGlu and isomaltose were 1.1 and 19.3 mM, respectively. CONCLUSION: Because of its unique properties, the α-glucosidase may have a potential in several industrial applications.

High-level production of ß-1,3-1,4-glucanase by Rhizomucor miehei under solid-state fermentation and its potential application in the brewing industry.

AIMS: To improve the ß-1,3-1,4-glucanase production by Rhizomucor miehei under solid-state fermentation (SSF) for industrial application. METHODS AND RESULTS: The fermentation conditions for ß-1,3-1,4-glucanase production by R. miehei CAU432 under SSF were optimized using a 'one-factor-at-a-time' method. Under the optimized fermentation conditions, viz. oatmeal (0·45-0·9 mm) as sole carbon source, 5% (w/w) peptone as sole nitrogen source, initial moisture of 80% (w/w), initial culture pH of 5·0, incubation temperature of 50°C and incubation time of 6 days, the highest ß-1,3-1,4-glucanase activity of 20,025 U g(-1) dry substrate was achieved, which represents the highest yield for ß-1,3-1,4-glucanase production ever reported. The crude enzyme was extracted and purified to homogeneity with a purification fold of 4·6 and a recovery yield of 9·0%. The addition of the purified ß-1,3-1,4-glucanase in mash obviously reduced its filtration time (24·6%) and viscosity (2·61%). CONCLUSIONS: The optimal fermentation conditions for maximal ß-1,3-1,4-glucanase production under SSF was obtained, and the enzyme was suitable for application in the malting process. SIGNIFICANCE AND IMPACT OF THE STUDY: The high production yield and excellent capability of the enzyme may enable it great potential in industries, especially in brewing industry.

Low energy-consumption plasma electrolytic oxidation based on grid cathode.

Plasma electrolytic oxidation (PEO) has attracted widespread attention owing to the simplicity of operation and the excellent properties of the formed coating. However, wider applications of PEO have been limited due to the high power consumption. This work describes the design and performance of a novel technique named shorter distance PEO (SD-PEO), which is intended for lowering the energy consumption. The key feature of the method is the application of grid cathode to eliminate the gaseous envelope effect and to block of the exchange of charge carries during SD-PEO process. Compared to PEO carried out at a normal electrode distance, e.g., 50 mm, both the voltage drop and the joule heat consumed in the electrolyte at a shorter distance, e.g., of 5 mm (SD-PEO) are relatively small. Consequently, the energy consumption rendered by the novel SD-PEO method may decrease by more than 25%. Our results reveal that SD-PEO is a low energy-consumption microarc oxidation technique with more potential in industry applications.

Simple and rapid detection of Salmonella serovar Enteritidis under field conditions by loop-mediated isothermal amplification.

AIM: The objective of this study is to develop a serovar-specific loop-mediated isothermal amplification (LAMP) method for sensitive, rapid, and inexpensive detection of Salmonella serovar Enteritidis under field conditions. METHODS: A set of six specific primers was designed with Salmonella Enteritidis DNA as the target. LAMP conditions were optimized by incubating the target DNA with the Bst DNA polymerase large fragment in a simple water bath. The sensitivity and specificity of LAMP was then compared with those of fluorescent quantitative real-time polymerase chain reaction (FQ-PCR). RESULTS: The results were as follows. (1) Serovar-specific Salmonella Enteritidis DNA was amplified at 65°C in as early as 20min in a water bath. (2) A colour change visible to the naked eye indicated a positive amplification reaction. (3) The detection limit of the LAMP assay was 4 copies µl(-1) ; thus, the sensitivity and specificity of this assay is similar to those of the FQ-PCR. CONCLUSIONS: LAMP is a high-throughput detection technique with high sensitivity, specificity, and simplicity; these factors make it suitable for specifically detecting Salmonella Enteritidis under field conditions and in laboratory settings. Thus, LAMP eliminates the need for complicated equipment and technical training in the detection of this specific serovar. SIGNIFICANCE AND IMPACT OF THE STUDY: This is the first study involving the use of LAMP to detect Salmonella serovar-specific DNA sequences. It is also the first to report an ideal method of distinguishing between Salmonella Enteritidis and other Salmonella under field conditions.

A dendritic cell-based tumour vaccine for lung cancer: full-length XAGE-1b protein-pulsed dendritic cells induce specific cytotoxic T lymphocytes in vitro.

XAGE-1b is regarded as one of the most immunogenic antigens and the most promising targets for lung adenocarcinoma immunotherapy. In this study, we sought to determine whether monocyte-derived dendritic cells (DCs) pulsed with purified full-length XAGE-1b could induce specific cytotoxic T lymphocytes (CTLs) against tumour cells from patients with non-small cell lung cancer (NSCLC) in vitro. XAGE-1b mRNA expression was examined in primary cultures of lung cancer cells and normal lung epithelial cells established from fresh tissues surgically resected from 30 patients with NSCLC using reverse transcription-polymerase chain reaction (RT-PCR). XAGE-1b mRNA expression was observed in 11 of 18 (61.1%) adenocarcinomas and one of 12 (8.3%) lung cancers of other histological types (P = 0.015). The 246-base pairs XAGE-1b gene was inserted into a recombinant expression vector. Full-length XAGE-1b was then expressed in BL21 (DE3) Escherichia coli and purified by AKTA-fast performance liquid chromatography (FPLC). DCs generated from peripheral blood mononuclear cells were pulsed with XAGE-1b by incubation with the protein at an immature stage. The XAGE-1b-pulsed DCs induced CTLs following 14 days of co-culture. Finally, an adherent target detachment (ATD) assay was performed to test the cytotoxicity of the XAGE-1b-specific CTLs against cancer cells and normal lung epithelial cells. The XAGE-1b-specific CTLs had a stronger lytic effect on autologous XAGE-1b mRNA-positive cancer cells than on autologous XAGE-1b mRNA-negative cancer cells or allogenous XAGE-1b mRNA-positive cancer cells. The CTLs had no lytic activity against normal lung epithelial cells. These results can be used to develop simple and effective cancer/testis antigen-based immunotherapies for NSCLC.

A xylose-tolerant beta-xylosidase from Paecilomyces thermophila: characterization and its co-action with the endogenous xylanase.

An extracellular beta-xylosidase from the thermophilic fungus Paecilomyces thermophila J18 was purified 31.9-fold to homogeneity with a recovery yield of 2.27% from the cell-free culture supernatant. It appeared as a single protein band on SDS-PAGE with a molecular mass of approx 53.5 kDa. The molecular mass of beta-xylosidase was 51.8 kDa determined by Superdex 75 gel filtration. The enzyme was a glycoprotein with a carbohydrate content of 61.5%. It exhibited an optimal activity at 55 degrees C and pH 6.5, respectively. The enzyme was stable in the range of pH 6.0-9.0 and at 55 degrees C. The purified enzyme hydrolyzed xylobiose and higher xylooligosaccharides but was inactive against xylan substrates. It released xylose from xylooligosaccharides with a degree of polymerization ranging between 2 and 5. The rate of xylose released from xylooligosaccharides by the purified enzyme increased with increasing chain length. It had a K(m) of 4.3mM for p-nitrophenol-beta-d-xylopyranoside and was competitively inhibited by xylose with a K(i) value of 139 mM. Release of reducing sugars from xylans by a purified xylanase produced by the same organism increased markedly in the presence of beta-xylosidase. During 24-hour hydrolysis, the amounts of reducing sugar released in the presence of added beta-xylosidase were about 1.5-1.73 times that of the reaction employing the xylanase alone. This is the first report on the purification and characterization of a beta-xylosidase from Paecilomyces thermophila.