Molecular and immunochemical characterization of profilin as major allergen from Platanus acerifolia pollen.

BACKGROUND: The Platanus acerifolia (P. acerifolia) pollen is one of the most common causes of allergic respiratory symptoms in China. However, the allergenic components in P. acerifolia are not fully studied yet. The study aimed to determine the molecular and immunochemical characterization of the profilin from P. acerifolia pollen. METHODS: The coding sequence of profilin was amplified, cloned, and then expressed in Escherichia coli BL21 cells and purified by nickel affinity chromatography. Protein refolding was followed by structural characterization and homology 3D model building. The allergenicity and cross-reactivity were assessed by ELISA, immunoblotting, or basophil activation test (BAT) using the sera of P. acerifolia allergic patients. RESULTS: The cDNA sequence of profilin was cloned with a 396 bp open reading frame coding for 131 amino acids. The molecular weight of the profilin was approximately 14 kDa, and the predicted structure consisted of 3 α-helixes and 7 ß-sheets. Physicochemical analysis indicated the profilin was a stable, relatively thermostable, and relatively conserved protein. The allergenicity determined by ELISA, western blot, and BAT suggested 76.9% (30/39) of the P. acerifolia pollen allergic patients displayed specific IgE recognition of the profilin. The profilin shared > 80% sequence identity with Pop n 2, the profilin from Populus nigra, and observed a significant cross-reactivity with Pop n 2 in IgE-inhibition assay. CONCLUSION: Profilin, as one of the major component allergens in P. acerifolia pollen, was identified and characterized at molecular and immunochemical levels in this study. These findings would contribute to developing diagnostic and therapeutic strategies for P. acerifolia pollen allergic patients.

Identification of Per a 13 as a novel allergen in American cockroach.

BACKGROUND: Cockroaches are an important source of indoor allergens. Environmental exposure to cockroach allergens is closely associated with the development of immunoglobulin E (IgE)-mediated allergic diseases. However, the allergenic components in the American cockroaches are not fully studied yet. In order to develop novel diagnostic and therapeutic strategies for cockroach allergy, it is necessary to comprehensively investigate this undescribed allergen in the American cockroach. METHODS: The full-length cDNA of the potential allergen was isolated from the cDNA library of the American cockroach by PCR cloning. Both the recombinant and natural protein molecules were purified and characterized. The allergenicity was further analyzed by enzyme linked immunosorbent assay, immunoblot, and basophil activation test using sera from cockroach allergic patients. RESULTS: A novel allergen belonging to glyceraldehyde 3-phosphate dehydrogenase (GAPDH) was firstly identified in the American cockroach and named as Per a 13. The cDNA of this allergen is 1255 base pairs in length and contains an open reading frame of 999 base pairs, encoding 332 amino acids. The purified Per a 13 was fully characterized and assessed to react with IgEs from 49.3 % of cockroach allergic patients, and patients with allergic rhinitis were more sensitized to it. Moreover, the allergenicity was further confirmed by immunoblot and basophil activation test. CONCLUSIONS: We firstly identified GAPDH (Per a 13) in the American cockroach, which is a novel type of inhalant allergen derived from animal species. These findings could be useful in developing novel diagnostic and therapeutic strategies for cockroach allergy.

Undiagnosed pleural effusion treated with traditional Chinese medicine: A case report.

The main clinical manifestations of pleural effusion are exertional dyspnea, predominantly dry cough, and pleuritic chest pain. To treat pleural effusion appropriately, it is important to determine its etiology; which however, remains unclear in nearly 20% of cases.A 73-year-old man with a history of invasive pulmonary tuberculosis (TB), had been experiencing chest congestion and dyspnea with undiagnosed pleural effusion for six years. After a series of clinical examination and laboratory tests, there was still no clear diagnosis. Despite administering diuretics and intermittent draining, the patient's condition aggravated progressively. He sought further treatment at Dongzhimen Hospital Respiratory Outpatient Clinic. The patient was treated with Zanthoxylum and Trichosanthes Decoction (Jiao Mu Gua Lou Tang). After one and a half years, his symptoms greatly improved and ultrasound revealed that the pleural effusion had apparently absorbed.It is suggested that TCM herbal formulas can play a critical role in preventing the progression of complicated, undiagnosed pleural effusion, especially in cases of poor response to conventional therapy and thoracentesis. Additional studies on the functions and mechanisms of the medicinals are warranted.

[Impact of a Sewage Treatment Plant on the Accumulation of Microplastics in Freshwater Organisms in the Lijiang River of the Guilin Urban Section].

Microplastics (MPs, particle size<5 mm), as a new pollutant, have attracted wide attention in recent years. The distributions of MPs in effluent of a sewage treatment plant (STP) were examined. Surface water, sediment, and freshwater organism samples were taken from the STP discharge outlet in the Lijiang River tributary (S1), the confluence of tributaries and main streams in the Lijiang River (S2), and downstream locations in the Lijiang River (S3). The impact of STP discharge effluent on the characteristics and spatial distribution of MPs pollution in freshwater organisms was studied. The results showed that the freshwater organisms had a probability of uptake of MPs by 94.2%. The mean abundance of MPs in S1 (2.7 n·ind-1) was significantly higher than that of S3 (1.9 n·ind-1, P<0.05). The MPs found in S1 and S3 were mainly <0.10 mm, accounting for 46.0% and 30.5%, respectively. The fiber type of MPs was observed in the body of freshwater organisms. Polyethylene terephthalate was the major polymer form in S1 organisms, while polypropylene was the major polymer form in S3. The effluent discharged from the STP led to the accumulation of MPs in freshwater organisms.

Shufeng Jiedu capsules for treating acute exacerbations of chronic obstructive pulmonary disease: a systematic review and meta-analysis.

BACKGROUND: Chinese herbal medicine is widely used in combination with usual care for acute exacerbations of chronic obstructive pulmonary disease (AECOPD) in China. Chinese patent medicine Shufeng Jiedu (SFJD) capsules is widely used for respiratory infectious diseases. This review aims to evaluate effectiveness and safety of SFJD for AECOPD. METHODS: A systematic review of randomised controlled trials (RCTs) in patients with AECOPD, who received SFJD as a single intervention or as add-on treatment to usual care. PubMed, the Cochrane Library, EMBASE, Scopus, Web of Science and four Chinese databases were searched from inception to April 2019. Two authors screened trials, extracted data, and assessed risk of bias, independently. Meta-analysis was performed using RevMan 5.3 software. We performed subgroup analyses and sensitivity analyses according to the predefined protocol. Quality of evidence was assessed using GRADE. RESULTS: Thirteen RCTs (1036 patients, with 936 inpatients) were included, all compared SFJD in combination with usual care (including antibiotics) to usual care alone. The mean age of participants ranged from 52 to 67 years, with approximately 60% male. Due to lack of blinding and other factors, all trials were of high risk of bias. SFJD was associated with a significant reduction in treatment failure, from 20.1 to 8.3% (11 trials; 815 patients; relative risk 0.43, 95% confidence interval [CI] 0.30 to 0.62), and duration of hospital stay (2 trials; 79 patients; mean difference - 4.32 days, 95% CI - 5.89 to - 2.75 days). No significant difference in adverse events was found between SFJD and control groups. CONCLUSION: Low certainty evidence suggests SFJD may bring additional benefit in reducing treatment failure, shorten hospital stay, and improving symptoms. Further large, high quality RCTs are needed to confirm its benefit and safety. TRIAL REGISTRATION: PROSPERO CRD42019133682.

Aerobic removal of iodinated contrast medium by nano-sized zero-valent iron: A combination of oxidation and reduction.

The removal performance and mechanisms of diatrizoate (DTA), a typical iodinated contrast medium, from water by nano-sized zero-valent iron (nZVI) under aerobic conditions were investigated in this study. Reactive oxygen species (ROS) and transformation products were detected with electron spin resonance and liquid chromatography electrospray ionization tandem mass spectrometry, respectively. Furthermore, the effects of several operational parameters on DTA removal were illustrated. The results showed that nZVI had a much higher DTA removal ability compared to microscale zero-valent iron (mZVI) in the presence of oxygen. Moreover, the detection of ROS and I- as well as the analysis of intermediate products suggested a combination of oxidation and reduction pathways for DTA removal by nZVI under aerobic conditions. Additionally, a high dosage of nZVI and acidic conditions led to the enhancement of DTA removal, while nZVI aging, as well as chloride and nitrate ions in the solution, had negative effects on the degradation of DTA by nZVI in the presence of oxygen.

Bioelectrochemical decolorization of a reactive diazo dye: Kinetics, optimization with a response surface methodology, and proposed degradation pathway.

Bioelectrochemical systems (BESs) have shown great potential for azo dye removal. However, comprehensive evaluation of the bioelectrochemical decolorization performance for reactive diazo dyes remains limited, particularly the kinetics and operation parameter optimization. This study evaluated the decolorization of the diazo dye Reactive Black 5 (RB5) in BESs, particularly with regard to kinetics, parameter optimization using response surface methodology (RSM), and the degradation pathway. The results indicated that the pseudo-first-order kinetic rate constant of RB5 decolorization increased from 0.023 ±â€¯0.001 to 0.146 ±â€¯0.008 h-1 with a decrease in cathode potential from -400 mV to -500 mV. RSM optimization suggested that the linear effects of RB5 concentration, cathode potential and hydraulic retention time (HRT), interaction of RB5 concentration with cathodic HRT, and the quadratic effect of cathodic HRT were most influential on the bioelectrochemical decolorization of RB5. Further, the decolorized RB5 products in the BESs were characterized by ultraviolet-visible spectrophotometry, Fourier-transform infrared spectroscopy, and liquid chromatograph-mass spectrometry. From this, a potential decolorization mechanism is proposed based on cleavage of azo bonds.

[Biomechanical research of antegrade intramedullary fixation for the metacarpal fractures].

OBJECTIVE: To study the biomechanical characteristics of antegrade intramedullary fixation for metacarpal fractures. METHODS: From March to May 2008, both the 4th and 5th metacarpals from 25 formalin embalmed cadaver hands had three-point bending test after transverse osteotomy followed by randomly fixation with one of the following three methods: plate and screw, antegrade intramedullary K-wire, crossed K-wire. While, both the 2nd and 3rd metacarpals had torsional loading test after the same management as the 4th and 5th metacarpal had undergone. RESULTS: In the three-point bending test, both the maximum bending moment (M(max)) and bending rigidity (EI) of the antegrade intramedullary K-wire were comparable with those of the plate and screw, and were significantly larger than those of the crossed K-wire. In the torsional loading test, the antegrade intramedullary K-wire had a statistically smaller maximum torque (T(max)) than the plate and screw, and had a comparable T(max) with the crossed K-wire; while, the torsional rigidity (GJ) of the intramedullary K-wire was statistically weaker than that of both the plate and screw and the crossed wire. CONCLUSIONS: One single antegrade intramedullary K-wire can provide a satisfactory M(max) and EI for metacarpal fixation and shows relatively weak in the torsional loading test. The injured finger should be well protected to avoid torsional deformity in clinical practice.

[Mutations in the D-loop region of mitochondrial DNA and the ROS level in the tissue of hepatocellular carcinoma].

To explore the relationship between ROS level and mutations in D-Loop region of mtDNA, mutations in the D-Loop region of mtDNA and the ROS level in primary hepatocarcinoma tissues were studied. We amplified the D-Loop region of mtDNA of 20 hepatocarcinomas and their adjacent tissue by PCR and then sequencing. ROS in tissue was measured by flow cytometry. mtDNA mutations were detected in 40% (8 of 20) tumor samples. 53 point mutations were detected in eight tumour samples, including 2 insertions, 11 deletions and 40 point mutations. 75% point mutations were T-C and C-T transition. They were four microsatellites among the mutations. Mutations in the adjacent tissues were always companied with mutations in tumour tissues. The mutation frequency in tumour tissues was higher than that in adjacent tissue. There was a larger unidentified deletion. The ROS level in hepatocarcinoma tissue was much higher than control (P<0.01). Meanwhile, we found the ROS level in hepatocarcinoma tissues with mutated mtDNA D-Loop was higher than that hepatocarcinoma tissue normal mtDNA D-Loop, and the ROS level in hepatocarcinoma adjacent tissue with mutated mtDNA D-Loop was higher than that in hepatocarcinoma adjacent tissue with normal mtDNA D-Loop. It was concluded that the D-Loop region of mitochondrial DNA was a highly polymorphoric and mutable region and mutation rate was relatively high in patients with hepaticellular carcinoma, and the abnormal ROS level might be the point mutation in the mitochondrial DNA and hepatocarcinogenesis related to ROS.

Herpes simplex virus thymidine kinase and ganciclovir suicide gene therapy for human pancreatic cancer.

AIM: To investigate the in vitro effects of suicide gene therapy system of herpes simplex virus thymidine kinase gene (HSV-TK) in combination with the treatment of nucleotide analog-ganciclovir (GCV) on human pancreatic cancer, and to provide a novel clinical therapeutic method for human pancreatic cancer. METHODS: We used a replication defective recombinant retrovirus vector GINaTK (bearing HSV-TK gene) to make packaging cell PA317 produce progeny virions. We then transferred the HSV-TK gene to target cells SW1990 using these progeny virions, and treated these gene-modified tumor cells with GCV to study the sensitivity of the cells to GCV and their bystander effects by routine MTT-method. RESULTS: Packaging cell PA317/TK was successfully constructed, and we acquired SW1990/TK through virus progeny infection. These gene-modified pancreatic cancer cells were sensitive to the treatment of GCV compared with unmodified tumor cells (t=4.15, n=10, P<0.0025). We also observed a remarkable bystander effect by mixing two kinds of cells at different ratio. CONCLUSION: Our data demonstrate that HSV-TK/GCV suicide gene therapy system is effective for treating experimental human pancreatic cancer, which is largely resistant to the common therapies, so the suicide gene therapy system may be a potential treatment approach for pancreatic cancer.

Spatiotemporal changes of the N-methyl-D-aspartate receptor subunit levels in rats with pentylenetetrazole-induced seizures.

The aim of this study was to examine the expression profiles of N-methyl-D-aspartate (NMDA) receptor subunits in rats during seizure development and kindled process induced by pentylenetetrazole (PTZ). Using quantitative Western blotting, the levels of NR1, NR2A and NR2B subunits were measured in the cortex and hippocampus of rats at different times after PTZ injection. In the early seizure developmental process, both NR1 and NR2A were markedly increased in the cortex, and NR1 was significantly increased in the hippocampus. On the other hand, in the kindled process both NR1 and NR2A decreased in the cortex and hippocampus. However, the NR2B subunit had no appreciable change in both the seizure developmental and kindled process. Therefore, these results showed that expression of NMDA receptors undergoes subunit- and region-related changes in the developmental and kindled seizure of rats induced by PTZ.

Relationship between telomerase activity and its subunit expression and inhibitory effect of antisense hTR on pancreatic carcinoma.

AIM: To directly investigate the relationship between telomerase activity and its subunit expression and the inhibitory effect of antisense hTR on pancreatic carcinogenesis. METHODS: We examined the telomerase activity and its subunit expression by cell culture, polymerase chain reaction (PCR), PCR-silver staining, PCR-ELISA, DNA sequencing, MTT and flow cytometry methods. RESULTS: PCR-silver staining and PCR-ELISA methods had the same specificity and sensitivity as the TRAP method. Telomerase activity was detected in the extract of the 10(th),20(th) and 30(th) passages of P3 cells,while it was absent in fibroblasts. Furthermore, after the 30th generation, the proliferation period of fibroblast cells was significantly prolonged. Telomerase activity and hTERTmRNA were detected in two pancreatic carcinoma cell lines, but were found to be negative in human fibroblast cells. Telomerase activity and hTERTmRNA were tested in pancreatic carcinoma specimens of 24 cases. The telomerase activity was positive in 21 of the 24 cases (87.5 %), and the hTERTmRNA in 20 cases (83.3 %). In adjacent normal tissues positive rates were both 12.5 %. There was a significant difference between the two groups. This indicated a significant correlation between the expression level of telomerase activity and histologic differentiation, metastasis and advanced clinical stage of pancreatic carcinoma. Our findings showed that the expressions of hTR and TP1mRNA were not correlated with the activity of telomerase but the expression of hTERTmRNA was. After treatment with PS-ODNs, telomerase activity in P(3) cells weakened and the inhibiting effect became stronger with an increase in PS-ODNs concentration. There was a significant difference between different PS-ODN groups (P<0.05). Inhibition of telomerase activity occurred most significant with PS-ODN1. The results of the FCM test of pancreatic cancer P(3) cells showed an increase in the apoptotic rate with increasing PS-ODN1 and PS-ODN2 concentrations. CONCLUSION: The expression of telomerase activity has a significant relationship to carcinogenesis. A strong correlation exists between telomerase activity and hTERTmRNA expression. The up-regulation of hTERTmRNA expression may play a critical role in human carcinogenesis. The expression of telomerase activity and its subunit level in pancreatic carcinoma significantly correlate with the clinical stage of pancreatic carcinoma and hence, may be helpful in its diagnosis and prognosis. The anti-hTR complementary to the template region of hTR is sufficient to inhibit P3 cell telomerase activity and cell proliferation in vitro, and can lead to a profound induction of programmed cell death.

[Inhibition effects of phosphorothioate-modified antisense hTR on the telomerase activity and the growth of P3 cells derived from pancreatic cancer in culture].

This paper is to investigate PS-ODN's (antisense-PS-ODN of hTR,sense-PS-ODN of hTR and random sequence) effects on telomerase activity and proliferation of P3 pancreatic cancer cells,and to find a novel method for gene therapy of pancreatic cancer. The results indicate that the anti-hTR complementary to the template region of hTR is sufficient to inhibit P3 cell telomerase activity and cell proliferation in vitro,and as a result, they can lead to a profound induction of programmed cell death. Telomerase represents an interesting and promising anticancer drug target and anti-telomerase technology may have potential significance in tumor therapy.