Urate oxidase (Uox) plays a pivotal role in uric acid (UA) degradation, and it has been applied in controlling serum UA level in clinical treatment of hyperuricemia (HUA). However, because Uox is a heterogenous protein to the human body, the immune rejections typically occur after intravenous administration, which greatly hampers the application of Uox-based agents. In this study, we used Lactococcus lactis NZ9000, a food-grade bacterium, as a host to express exogenous Uox genes, to generate the Uox-expressing engineered strains to treat HUA. Aspergillus flavus-derived Uox (aUox) and the "resurrected" human-derived Uox (hUox) were cloned into vector and expressed in NZ9000, to generate engineered strains, respectively. The engineered NZ9000 strains were confirmed to express Uox and showed UA-lowering activity in a time-dependent manner in vitro. Next, in an HUA mice model established by oral gavage of yeast paste, the UA levels were increased by 85.4% and 106.2% at day 7 and day 14. By contrast, in mice fed with NZ9000-aUox, the UA levels were increased by 39.5% and 48.3% while in mice fed with NZ9000-hUox were increased by 57.0% and 82.9%, suggesting a UA-lowering activity of both engineered strains. Furthermore, compared with allopurinol, the first-line agent for HUA treatment, mice fed with NZ9000-aUox exhibited comparable liver safety but better kidney safety than allopurinol, indicating that the use of engineered NZ9000 strains not only alleviated kidney injury caused by HUA, but could also avoided the risk of kidney injury elicited by using allopurinol. Collectively, our study offers an effective and safe therapeutic approach for HUA long-term treatment and controlling.
Animals ; Lactococcus lactis/metabolism* ; Urate Oxidase/genetics* ; Mice ; Uric Acid/blood* ; Hyperuricemia ; Humans ; Administration, Oral ; Aspergillus flavus/genetics* ; Male

Objective:To investigate the safety and efficacy of MR-guided focused ultrasound surgery (MRgFUS) in treating localized prostate cancer.Methods:Six patients with localized prostate cancer who underwent MRgFUS treatment from August 2020 to September 2021 in Beijing Hospital were prospectively enrolled in this study. The patients were all over 18 years old, with an average age of (68±10) years, and had not received any prior treatment for prostate cancer. Pretreatment pelvic MR and CT scans were performed to determine the region of treatment (ROT). Different urinary catheterization measures were given based on the location of the lesion. Under general anesthesia, the lesions were treated by MRgFUS using a specialized rectal ultrasound transducer on the treatment bed. The patients were followed up at 1, 3, and 6 months after treatment and annually thereafter. During follow-up, prostate-specific antigen (PSA) levels, pelvic MR scans, International Prostate Symptom Score (IPSS), International Index of Erectile Function-15 (IIEF-15) scores, and adverse events were assessed.Results:(1) All six patients underwent MRgFUS treatment for six lesions, with an average duration of (126±56) minutes, an average number of (7.3±3.2) focal ultrasound pulses per lesion, and an average non-perfusion volume of (3.8±1.1) cm 3, which covered the entire treatment target area. No treatment-related adverse events were reported. (2) The PSA levels at baseline, 1, 3, 6, and 12 months after treatment were (6.6±0.8), (3.6±1.3), (3.4±3.0), (2.5±1.7), and (2.3±1.8) ng/ml, respectively. PSA levels increased in 2 out of 6 patients during follow-up, and pelvic MR scan revealed recurrent lesions, while PSA levels continued to decrease in the remaining 4 patients, and pelvic MR scan were normal. (3) The IPSS scores at baseline, 1, and 3 months after treatment were 13.0 (4.0, 16.0), 10.0 (4.0, 12.0), and 5.0 (3.0, 6.0) points, respectively. For the three sexually active patients, the IIEF-15 scores at baseline were 40, 51, and 14 points, respectively, and IIEF-15 at 1 month after treatment were 9, 8, and 14 points, respectively, and at 3 months after treatment were 9, 66 and 26 points, respectively. (4) One patient was diagnosed with recurrence 10 months after treatment, and another patient was found to have a new lesion 6 months after treatment. Conclusions:MRgFUS might be a safe, non-invasive, and effective treatment for localized prostate cancer, but regular follow-up is vital for detecting tumor recurrence.

Objective:To investigate the application value of dual-energy CT in the differential diagnosis of lung metastases and benign nodules in breast cancer.Methods:The data of 96 patients with pathology-confirmed breast cancer at the Fifth Affiliated Hospital of Wenzhou Medical University from March 2017 to June 2021 were analyzed retrospectively. All patients received dual-energy chest CT scans within 2 weeks before surgery. All 96 patients were female, aged 31-84 (56±12) years. A total of 207 pulmonary nodules from 96 patients were classified into 81 lung metastases and 126 benign nodules according to pathological findings. Conventional CT features [longest diameter, boundary, location and CT value difference between arterial and venous phases (ΔCT) of nodules] and dual-energy CT parameters [standardized iodine concentration (NIC), slope of energy spectrum (λ HU) and normalized effective atomic number (nZ eff) in arterial and venous phases] were analyzed and measured. The χ 2 test, independent samples t test and Kruskal-Wallis rank-sum test were used to analyze the differences of conventional CT features and dual-energy CT parameters between lung metastases and benign nodules. First, the least shrinkage and selection operator (LASSO) regression method was used to screen conventional CT features and dual-energy CT parameters, and then logistic regression analysis was performed to screen out independent risk factors for lung metastases. Receiver operating characteristic (ROC) curves were used to evaluate the efficacy of CT parameters alone and logistic model in differentiating lung metastases from benign lung nodules. Results:There were statistically significant differences between lung metastases and benign nodules in longest diameter, ?CT, NIC, λ HU and nZ eff in arterial and venous phases (all P<0.05). LASSO regression and binary logistic regression analysis showed that the venous phase λ HU (OR=59.413, 95%CI 14.233-248.002, P<0.001) and the venous phase nZ eff (OR=4.508, 95%CI 2.787-7.290, P<0.001) were independent risk factors for predicting lung metastases. Among them, the venous phase λ HU had the highest diagnostic efficiency, with an area under curve (AUC) of 0.794 and an accuracy of 74.88%. The AUC of the logistic model constructed by combining the venous phase λ HU and the venous phase nZ eff could reach 0.958, and the accuracy was improved to 92.27%, which was significantly higher than the efficacy of the two alone ( Z=6.02, 9.54, all P<0.001). Conclusion:Dual-energy CT has great application value in the identification of lung metastases and benign nodules in patients with breast cancer, especially when combined with venous phase λ HU and venous phase nZ eff, the diagnostic efficiency is further improved.

Objective:To evaluate the efficacy and safety of lenalidomide plus dexamethasone (LD) in patients with proliferative glomerulonephritis with monoclonal immunoglobulin deposits (PGNMID).Methods:The clinicopathological data of PGNMID patients who were treated with LD protocol from January 2010 to October 2019 were retrospectively analyzed.Results:All of 6 patients received LD treatment for≥3 months after renal biopsy in Jinling Hospital. During the follow-up period of 6 to 19 months, 3 patients achieved renal remission, and the renal remission rate was 50%(3/6). Light microscopy showed membranoproliferative glomerulonephritis and immunofluorescence showed single kappa type IgG3 was deposited in the mesangial region and the vascular loop. Before taking LD scheme, the median urinary protein were 7.76(1.27, 14.57) g/24 h, the median serum creatinine was 118.5(70.7, 289.1) μmol/L, and the median albumin was 34.5(22.4, 37.5) g/L. The concentration of serum free kappa and lambda light chain was increased in 5 patients, but the serum free light chain ratio was normal. Hypocomplementemia was detected in two cases. Six patients underwent bone marrow flow cytometry, and 2 patients had elevated monoclonal plasma cells, accounting for 0.7% and 0.5%, respectively. Immunofixation electrophoresis suggested that 1 patient had positive serum M protein for kappa type IgG3. At the last follow-up, median urine protein was 3.33(0.33, 11.23) g/24 h, median serum creatinine was 108.7(80.4, 160.9) μmol/L, and median albumin was 35.9(24.5, 45.6) g/L. The concentration of serum free light chain in 4 patients from 5 patients with elevated serum free light chain was lower than that before taking the drug. Decreased level of serum complement in two cases returned to normal after treatment. The M spike did not turn negative during the follow-up in one patient. Adverse events included anemia, neutropenia, limb numbness and upper respiratory tract infection.Conclusion:This study reports for the first time that LD protocol may be effective in treating PGNMID, but more attention should be paid to the hematological adverse events of lenalidomide.

DNA, Neoplasm ; Humans ; Image Cytometry ; Pleural Effusion, Malignant ; diagnosis

Objectives To explore the pathological characteristics, treatment and prognosis of medulloblastoma (MB) in children. Methods Pathological characteristics, treatment outcomes and other clinical data were retrospectively analyzed in 60 children with MB. Results The MB in all 60 children were high-grade tumor. The pathological type was mainly classic (45 cases, 75.00%) and other subtypes were seen in 15 cases (25.00%). Forty-eight (80.00%) children had total resection by micro-surgery, 9 children (15.00%) subtotal resection, and 3 children (5.00%) partial resection. Twenty-seven children were treated by radiotherapy after surgery, 6 children were treated by chemotherapy after surgery, and 5 children were treated with radiotherapy combined chemotherapy after surgery. Thirty-eight cases were followed up. The 2-year survival and disease-free survival rates were 63.16%and 55.26%. The 5-year survival and disease-free survival rates were 26.32%and 18.42%. The children with des-moplastic/nodular type survived significantly longer than the children with large cell anaplastic. The children with total resection survived significantly longer than the children with partial resection. The children with radiotherapy or chemotherapy survived significantly longer than the children without chemotherapy (P<0.05). Conclusions MB in most of children is high-grade tu-mors and the prognosis is poor. The pathological type is related with prognosis. Microsurgical total resection is the first choice of treatment. Postoperative radiotherapy and chemotherapy can increase the survival rate.

Background Fungal corneal ulcer is a visual-threatening eye disease,and drug therapy has a limiting efficacy.Corneal transplantation or eye enucleation sometimes is necessary to the severe patients.Corneal collagen cross-linking (CXL) is an effective method for some corneal diseases,but the study on CXL for fungal corneal ulcer is lack.Objective This study was to evaluate the clinical effectiveness and safety CXL for fungal corneal ulcer.Methods Fifteen 8-week-old healthy New Zealand white rabbits were used in this study and other 5 rabbits served as normal controls.Fungal corneal ulcer models were established in the right eyes of other 10 rabbits by infecting sickle bacteria liquid after corneal scratching and removing corneal epithelium,then decellularized ostrich corneal patch covered the defected cornea.The models were randomly divided into the non-treatment group and the CXL treatment group.Corneal lesions were examined under the slit lamp microscope every day,and cornea was pictured by laser scanning confocal microscope on the 3rd,7th,14th,21st and 28th day individually after CXL.All rabbits were sacrificed and corneal tissues were obtained 4 weeks after treatment,and the collagen fiber diameter and fibrocytes were observed under the scanning electron microscope.Results Fungal corneal ulcer models were successfully established by corneal scratching and decellularized ostrich cornea covering.The gray ulcer lesions and hypbae like bean pod were seen by slit lamp microscope and laser scanning confocal microscope 3 days after modeling.Corneal ulcer deepened and expanded 1 week later,and there were a large number of spore and hyphae criss-crossing as short rod in shallow stroma.Inflammatory cells were observed in corneal endothelial cells and ocular anterior chamber.In the CXL treatment group,the range of corneal epithelial deficiency was less than that in the nontreatment group on the 3rd,7th,14th,and 21st (all at P＜ 0.05).The diameters of collagen fibers were (24.6± 1.8) nm,(24.9 ± 1.9) nm and (43.0 ± 7.4) nm in the normal control group,non-treatment group and CXL treatment group,showing a significant difference among the 3 groups (F =27.05,P =0.00),and the collagen diameters were thicker in the CXL treatment group than those in the normal control group and non-treatment group (t =5.40,-5.30,both at P＜0.05),and fibrocytes were seen among the collagen fibers.No significant difference was found in the collagen diameters between the non-treatment group and normal control group,and the fibrocytes were less in the non-treatment group.Conclusions CXL therapy can treat fungal corneal ulcer by enhancing collagen,promoting fibrocytes proliferation,suppressing fungus and inflammatory response and accelerating tissue repair.

Chemical investigatation of Drosera peltata var. multisepala led to the isolation of eleven compounds using various chromatographic techniques. The structures of these compounds were elucidated as isoshinanolone-4-O-beta-D-glucoside (1), isoshinanolone (2), epi-isoshinanolone (3), plumbagin (4), droserone (5), droserone-5-O-glucoside (6), quercetin (7), kaempferol (8) , gossypetin-8-O-glucoside (9), 3,3'-dimethoxy ellagic acid (10), and ellagic acid (11) by their physicochemical properties and spectral data analysis. Compound 1 was a new compound. Compounds 3, 8, 10, and 11 were isolated from this plant for the first time.
Chromatography, Liquid ; methods ; Drosera ; chemistry ; Drugs, Chinese Herbal ; chemistry ; Ellagic Acid ; analogs & derivatives ; analysis ; isolation & purification ; Glucosides ; analysis ; isolation & purification ; Kaempferols ; analysis ; isolation & purification ; Magnetic Resonance Spectroscopy ; methods ; Molecular Structure ; Naphthoquinones ; analysis ; isolation & purification ; Plant Extracts ; analysis ; isolation & purification ; Quercetin ; analysis ; isolation & purification ; Spectrometry, Mass, Electrospray Ionization ; methods ; Tetrahydronaphthalenes ; analysis ; isolation & purification

Objective To study the effects of rehabilitation training on learning and memory ability and the expression of growth-associated protein-43(GAP-43)in rats with vascular dementia.Methods Forty-four female Sprague-Dawley rats were randomly assigned to a rehabilitation group(n=20),an immobilization group(n=20),and a sham-operation group(n=4).The rats in the former 2 groups were operated on to establish the experimental vascular dementia model by repeatedly ischemia/reperfusion injury of brain induced by ligation of bilateral common carotid arteries and lowering of blood pressure induced by intraabdominal injection of sodium nitroprusside.The rats in rehabilitation group were administered with rotating bar and rolling cage exercises for 1 hour once daily,while those in the immobilization group were immobilized and without any exercise:the rats in sham-operation group could move freely in cage.Learning and memory tests were preformed by using step-down avoidance test at the 27th and 28th days after operation.Immunohistochemical staining was used to detect GAP-43 expression in hippocampus CA1 area at different time points after operation. Results The rats in rehabilitation group demonstrated better learning and memory ability than those in immobilization group(P<0.01),and more GAP-43 expression in hippocampus CA1 ar-ea than those in immobilization group and sham-operation(P<0.01).Conclusion Rehabilitation training can im-prove the learning and memory ability of rats with experimental vascular dementia,and the mechanism is probably re-lated to the increase of GAP-43 in hippocampus CA1 area.