[Challenges and strategies in the treatment of neovascular age-related macular degeneration].

Neovascular age-related macular degeneration (nAMD) is a leading cause of blindness in the elderly, and anti-vascular endothelial growth factor (VEGF) therapy is currently the primary treatment approach. However, the real-world effectiveness of nAMD treatment is not always satisfactory and faces various challenges. Frequent administration and follow-up burdens can lead to decreased patient compliance during long-term treatment, resulting in suboptimal outcomes. Some lesions exhibit poor or no response to anti-VEGF treatment, leading to difficulties in maintaining or even declining visual acuity. Factors such as lesion fibrosis and tissue atrophy can contribute to visual deterioration. Therefore, standardizing and individualizing treatment plans, along with enhancing comprehensive monitoring and management throughout the disease course, are crucial improvement measures. The evidence-based guidelines for diagnosis and treatment of age-related macular degeneration in China, released in 2023, provide guidance for standardized clinical diagnosis and treatment. Meanwhile, research and development of new drugs and administration methods are anticipated for the future.

[Establishment of a prognostic model for non-nephrotic membranous nephropathy based on unbalanced data].

Objective: To explore the construction of a machine learning model based on unbalanced data to predict the progression of non-nephrotic membranous nephropathy. Methods: The clinical and pathological data of patients diagnosed with non-nephrotic membranous nephropathy by renal biopsy in Shanxi People's Hospital from January 2018 to December 2021 were retrospectively analyzed.The prediction models were constructed based on logistic regression, support vector machine (SVM) and light gradient boosting machine (lightGBM), respectively. The mixed sampling technology was used to process the unbalanced data, and the area under the receiver operating characteristic curve (AUC) was used to evaluate the predictive performance of the models. Finally, Shapley additive explanation (SHAP) was used to interpret the results of the optimal prediction model. Results: A total of 148 patients were included in the study, including 84 males and 64 females, with a mean age of (47.2±12.5) years. The follow-up time [M(Q1, Q3)] was 14(7, 20) months. Twenty-three patients (15.5%) achieved the renal end-point event in the study. The SVM model had the highest AUC (0.868, 95%CI: 0.813-0.925), followed by logistic regression (AUC=0.865, 95%CI: 0.755-0.899) and lightGBM (AUC=0.791, 95%CI: 0.690-0.882). The feature recursive elimination cross validation (RFECV) method based on random forest (RF) and the SHAP plot based on the SVM model showed that immunohistochemistry IgG, total protein (TP), anti-phospholipase A2 receptor (anti-PLA2R), blood chloride and D-Dimer were risk factors affecting the progress of non-nephrotic membranous nephropathy. Moreover, patients with high immunohistochemistry IgG, anti-PLA2R and D-Dimer had an increased risk of achieving the renal end-point event. Conclusion: The SVM model established in this study can effectively predict the progress of non-nephrotic membranous nephropathy, and provide a new method for the early identification of high-risk patients and precision therapy.

[Investigation of serum surface active protein D and clara cell protein levels in workers exposed to silica dust in ferrous metal foundry].

Objective: To investigate the levels of serum surface active protein D (SP-D) and clara cell protein (CCl6) in workers exposed to black silica dust, and analyze its influencing factors. Methods: From July to September 2021, 174 workers in 37 positions exposed to silica dust in 5 ferrous metal foundry were investigated by cross-sectional research method. The exposure concentration of silica dust workers was obtained through occupational health field investigation and detection, and the general situation of the study subjects was obtained through questionnaire survey and peripheral blood was collected. Double antigen sandwich enzyme-linked immunosorbent assay (ELISA) was used to detect the concentrations of SP-D and CC16 in serum of workers. The mean values were compared by one-way ANOVA, and the influencing factors of SP-D and CC16 concentrations in serum were analyzed by ordered multiple logistic regression. Results: The time-weighted average concentration (C-TWA) of 174 workers exposed to silica dust (respirable dust) ranged from 0.09 mg/m(3)~3.58 mg/m(3), and the C-TWA overstandard rate of dust exposed workers was 32.18% (56/174) , with differences among workers in different positions (χ(2)=28.85, P<0.001) . The highest concentration of silica dust was (0.82±0.11) mg/m(3). Using C-TWA<50% OEL occupational exposure limit (OEL) as reference, serum SP-D concentration in workers with ≥50% OEL was increased (OR=4.95, 95%CI: 1.86~13.17, P=0.001) , while CC16 concentration was decreased (OR=0.15, 95%CI: 0.05~0.40, P<0.001) ; Serum CC16 concentration decreased in workers exposed to silica dust C-TWA≥OEL (OR=0.46, 95%CI: 0.28~0.98, P=0.043) . Compared with those with low occupational health literacy, the serum SP-D concentration of workers with high occupational health literacy decreased (OR=0.48, 95%CI: 0.25~0.92, P=0.027) and CC16 concentration increased (OR=2.09, 95%CI: 1.10-3.97, P=0.024) . Conclusion: When no abnormality was found in the physical examination of workers, the serum SP-D and CC16 concentration levels changed, and the change was related to the concentration of workers exposed to silica dust.

[D-dimer contributes to the diagnosis and prognosis in hepatitis B-related acute-on-chronic liver failure].

Objective: To investigate the diagnostic and prognostic value of D-dimer level in patients with hepatitis B-related acute-on-chronic liver failure (HBV-ACLF). Methods: A total of 142 cases diagnosed with ACLF were randomly selected as research objects in the open cohort using the Chinese Group on the Study of Severe Hepatitis B-ACLF (COSSH-ACLF). Plasma D-dimer levels were compared between patients with ACLF and non-ACLF and patients with different ACLF grades. Survival and death group D-dimer levels were compared with the end points of 28 days and 90 days, respectively. The correlation between D-dimer and other laboratory indicators and prognostic scores were investigated. Area under receiver operating characteristic curve (AUROC) was used to evaluate the D-dimer value for predicting the prognosis of ACLF patients. 125 external ACLF cases were used for validation. A Student t test or Mann-Whitney U test was used to compare continuous measurement data between two groups. Kruskal-Wallis test was used to compare continuous measurement data between multiple groups. Results: Plasma D-dimer levels in the ACLF [2 588.5 (1 142.8, 5 472.8) µg/L] ] and non-ACLF group [1 385.5 (612.0, 3 840.3) µg/L] had a significant difference (P<0.001). ACLF-3 patients had significantly higher D-dimer levels than ACLF-1/2 patients (ACLF-3 vs. ACLF-1, P<0.001; ACLF-3 vs. ACLF-2, P<0.05). Patients who died at 28/90 days had significantly higher D-dimer levels than those whom survived (P<0.001). There was a significant positive correlation between D-dimer level with prothrombin time (PT), international normalized ratio (INR), high-density lipoprotein C, as well as various prognostic scores (COSSH-ACLFs, CLIF-C ACLFs, CLIF-OFs, MELDs). AUROC of D-dimer in predicting the prognosis of ACLF patients at 28 days and 90 days was 0.751 (95% CI: 0.649-0.852) and 0.787 (95% CI: 0.695-0.878), respectively, which did not differ significantly compared with the predictive ability of other scores (P<0.05), and similar results were confirmed by an external validation group of 125 cases. Conclusion: D-dimer level is significantly higher in patients with ACLF, so it is an independent predictor of prognosis at 28 and 90 days.

[A meta-analysis on prevalence of primary angle-closure glaucoma in the middle-aged and elderly chinese population].

Objective: To evaluate the prevalence and relevant factors of primary angle-closure glaucoma (PACG) in the middle-aged and elderly (40 years old and above) Chinese population. Methods: Epidemiological literature (of English and/or Chinese language) on prevalence of PACG in China between 2000 and 2022 were retrieved from PubMed, Embase, Web of Science, CNKI, and Wanfang database. Two researchers conducted the process of document retrieval, inclusion, quality assessment and data extraction independently based on inclusion and exclusion criteria. Disagreement was resolved by consensus with a third researcher. The heterogeneity among studies was assessed by using the I2 statistics, and a corresponding model was used for the meta-analysis. Subgroup analysis (gender, age, survey date, place of residence, diagnostic criteria, and temperature zone), as well as sensitivity analysis and publication bias analysis, were conducted. Results: Twenty-three studies were included in this research. Among them, eight were of high-quality and fifteen were of middle-quality. In the past 20 years, the PACG prevalence in middle-aged and elderly (40 years and older) Chinese population was 1.35% (95%CI:1.08% to 1.61%). Chinese women (1.40%) were more likely to have PACG than men (0.96%, P<0.01). The PACG prevalence had a positive correlation with age (P<0.01). The prevalence was similar between urban (1.3%) and rural (1.2%, P=0.61) areas. The PACG prevalence from studies in which the diagnostic criteria of PACG requested the symptom of vision or optic nerve damage (1.1%) was lower than that those without such requirement (1.7%, P<0.01). In mid-temperate zone, the prevalence was higher (1.7%, P=0.02). Conclusions: The PACG prevalence in middle-aged and elderly (40 years old and above) Chinese population was 1.35% in the last 20 years. The prevalence of PACG varies across age, regions, and other variations. This risk is higher in the elderly, women, and people in cold regions among Chinese population.

A novel bacteriocin BM1029: physicochemical characterization, antibacterial modes and application.

AIM: Bacteriocins with antimicrobial activity are considered as potential natural bio-preservatives to control the growth of food spoilage bacteria. The aim of this work was to characterize a novel bacteriocin BM1029 discovered from Lactobacillus crustorum MN047 and evaluate its antibacterial mechanism. METHODS AND RESULTS: Bacteriocin BM1029 was purified by cation-exchange chromatography and reversed-phase chromatography. Antibacterial activity assay showed that BM1029 is antagonistic against both Gram-positive and Gram-negative bacteria. Furthermore, it was found that BM1029 showed low haemolysis with high stability to the pretreatment with different temperatures, pH and surfactants. Moreover electron microscopy and flow cytometry suggested that BM1029 inhibit indicator strains by damaging the cell envelope integrity. Cell cycle assay suggested that BM1029 arrested cell cycle in R-phase. CONCLUSION: The novel bacteriocin BM1029 showed high bactericidal activity against Escherichia coli and Staphylococcus aureus through a cell envelope-associated mechanism. SIGNIFICANCE AND IMPACT OF THE STUDY: Application of BM1029 inhibited the growth of indicator strains on beef meat storage at 4°C suggesting that this bacteriocin is promising to be used as a novel preservative in food processing and preservation.

[Research progress of neural precursor cells-expressed developmentally down-regulated protein-8 in liver diseases].

Neural precursor cells-expressed developmentally down-regulated protein-8 (NEDD8) is one of the important members of the ubiquitin family, which plays an important role in maintaining cell stability, cell cycle regulation, signal transduction, transcription, and translation, DNA repair, and tumorigenesis through covalently bound substrates (also known as neddylation modification). In recent years, studies have found that the dysfunction of NEDD8 and its related enzymes is common in liver diseases, and is widely involved in the biological processes of hepatitis, liver fibrosis, proliferation, invasion, apoptosis and autophagy of liver cancer cells. This article focuses on the research progress of NEDD8 in liver diseases.

[Expression and diagnostic value of macrophage inflammatory protein-3α in patients with hepatitis B-related acute-on-chronic liver failure].

Objective: To screen serum protein markers and evaluate their diagnostic application value in hepatitis B-related acute-on-chronic liver failure (HBV-ACLF). Methods: Serum samples of patients with HBV-ACLF, chronic hepatitis B (CHB) and normal healthy volunteers (n = 5/group) were determined by cytokine antibody chip in line with the Chinese Diagnostic Standards Study for HBV-ACLF (COSSH-ACLF) cohort. The differentially expressed proteins significance were identified by microarray analysis and prediction. The preliminary serological markers of HBV-ACLF were screened for diagnosis. The potential markers were determined by enzyme-linked immunosorbent assay (ELISA), area under the receiver operating characteristic curve (AUROC) analysis and liver tissue immunohistochemistry for the diagnosis of HBV-ACLF. Student t-test or Mann-Whitney U test were used to compare the continuous measurement data between the two groups, and analysis of variance and Kruskal-Wallis test were used to compare the continuous measurement data between multiple groups. Results: Cytokine antibody chip preliminary screening results showed that the expression levels of these six cytokines, namely, macrophage inflammatory protein 3α (MIP-3α), hepatocyte growth factor, E-selectin, osteopontin, growth differentiation factor 15 and carcinoembryonic antigen-related cellular adhesion molecule 1 were significantly increased in the HBV-ACLF group. Among them, the expression level of MIP-3α was significantly higher in the HBV-ACLF group (99.6 times higher than CHB group and 146.9 times higher than healthy volunteers' group, respectively, P < 0.0001) as validated by serum ELISA in 132 HBV-ACLF cases, 91 CHB cases and 72 healthy volunteers. AUROC analysis showed that the high expression of MIP-3α could be used as a marker to distinguish patients with HBV-ACLF from CHB. The AUROC was 0.995 (95% CI: 0.990 ~ 1.000), with sensitivity and specificity of 95.5% and. 98.9%, respectively. Immunohistochemistry showed that MIP-3α was positively expressed in HBV-ACLF-derived liver tissues, and negatively expressed in CHB-derived liver and normal liver tissues. Conclusion: Serum MIP-3α level is closely related to the pathological characteristics of HBV-ACLF. Therefore, it may be used as a potential serological marker for the diagnosis of HBV-ACLF.

[Application of dural puncture epidural technique for labor analgesia].

Objective: To investigate the effects of dural puncture epidural technique for labor analgesia on mothers and neonates. Methods: From January to June 2019, one hundred healthy and nulliparous women, scheduled for elective labor analgesia in the Second Affiliated Hospital of Wenzhou Medical University, met inclusion criteriaand were recruitedin this prospective study. The inclusion criteria are as follows: American Society of Anesthesiologists physical statusⅠorⅡ, New York Heart Association gradeⅠorⅡ,150-175 cm in height,50-90 kg in weight and 37-45 weeks of gestation. They were randomly divided into epidural analgesia group(group P, n=50)and dural puncture epidural group(group D, n=50) by using random number table. Parturients in group D received epidural catheterization immediate after successful epidural puncture, while parturients in group P received a single dural puncture into subarachnoid space with a 27 gauge needle (successful puncture: outflow of cerebrospinal fluid) before epidural catheterization. Epidural labor analgesia was performed with epidural infusion of 0.1% ropivacaine plus 0.25 µg/ml sufentanil in both groups. The VAS scores were evaluated at the following time points: before epidural infusion, each uterine contraction within 30 min after infusion, 30 min, 60 min and 90 min after infusion and withdrawal of infusion. Labor process, mode of delivery, cases of increased oxytocin using, effective PCA pressings, sufentanil and ropivacaine dosages, complications of analgesia, neonatal status were recorded, as well. Results: There were no significant differences in labor duration, mode of delivery, analgesia complications (nausea and vomiting, itching, headache after delivery and Bromage score for motor block), deceleration of fetal heart rate and neonatal Apgar score between the two groups (P>0.05). The number of effective PCA pressings, sufentanil dosage, ropivacaine dosage and cases of increased using of oxytocin were significantly more in group P(t=8.663,7.024,6.509,χ(2)=4.159,all P<0.05), with (8.6±2.5) times, (29±4) µg,(105±15) mg,28% in group P, compared with (4.6±2.1) times,(23±4) µg,(88±12) mg,10% in group D, respectively. The first four VAS scores of uterine contraction after analgesia in group P(VAS=7.9±1.1,6.8± 0.9, 5.6±0.8, 4.5±0.8)were significantly higher than those in group D (VAS=6.8±0.7,4.7±0.8,3.5±0.8,2.9±0.7,t=5.966,12.332,13.125,10.643,all P<0.05). The VAS scores at 90 min after analgesia and withdrawal of analgesia (VAS=2.7±0.6, 2.9±0.7) in group P were significantly higher than those in group D (VAS=2.4±0.6, 2.5±0.6, t=2.500, 3.068, all P<0.05). Conclusion: Compared with traditional epidural technique, dural puncture epidural technique can provide a rapid and effective analgesia with less analgesics, but without increasing adverse effects on mother and infant.

CRISPR/Cas9-mediated knockout of the eye pigmentation gene white leads to alterations in colour of head spots in the oriental fruit fly, Bactrocera dorsalis.

The intensely studied white gene is widely used as a genetic marker in Drosophila melanogaster. Here, we cloned and characterized the white gene in an important pest of the fruit industry, Bactrocera dorsalis, to understand its functional role in pigmentation. We obtained BdWhite knockout strains, based on the wild-type strain, using the CRISPR/Cas9 genome editing system, and found that mutants lost pigmentation in the compound eye and their black head spots. We then examined differences in the expression levels of genes associated with melanin pigmentation between mutants and the wild-type strain using quantitative reverse transcription PCR. We found that transcription levels of the Bd-yellow1 were lower in the head of mutants than in the wild-type strain, and there were no significant differences in expression of the other six genes between mutants and the wild type. Since yellow is critical for melanin biosynthesis (Heinze et al., Scientific Reports. 2017;7:4582), the lower levels of expression of Bd-yellow1 in mutants led to reduced dark pigmentation in head spots. Our results provide the first evidence, to our knowledge, that white may play a functional role in cuticle pigmentation by affecting the expression of yellow.

Intra- and inter-specific variation in alarm pheromone produced by Solenopsis fire ants.

Some fire ants of the genus Solenopsis have become invasive species in the southern United States displacing native species by competition. Although the displacement pattern seems clear, the mechanisms underlying competitive advantage remain unclear. The ability of ant workers to produce relatively larger amount of alarm pheromone may correspond to relative greater fitness among sympatric fire ant species. Here we report on quantitative intra-specific (i.e. inter-caste) and inter-specific differences of alarm pheromone component, 2-ethyl-3,6-dimethylpyrazine (2E36DMP), for several fire ant species. The alarm pheromone component was extracted by soaking ants in hexane for 48 h and subsequently quantified by gas chromatography-mass spectrometry at single ion monitoring mode. Solenopsis invicta workers had more 2E36DMP than male or female alates by relative weight; individual workers, however, contained significantly less pyrazine. We thus believe that alarm pheromones may serve additional roles in alates. Workers of Solenopsis richteri, S. invicta, and hybrid (S. richteri × S. invicta) had significantly more 2E36DMP than a native fire ant species, Solenopsis geminata. The hybrid fire ant had significantly less 2E36DMP than the two parent species, S. richteri and S. invicta. It seems likely that higher alarm pheromone content may have favored invasion success of exotic fire ants over native species. We discuss the potential role of inter-specific variation in pyrazine content for the relationship between the observed shifts in the spatial distributions of the three exotic fire ant species in southern United States and the displacement of native fire ant species.

[Advances in research on cell fate determinant Numb regulating liver cancer].

Liver cancer is one of the most common malignant tumors in China, ranking fifth in malignant tumors and the third in tumor-related deaths. As a membrane-related protein, the asymmetric distribution of cell fate determinant Numb plays a key role in cell differentiation. Research reports that Numb may be closely associated to the occurrence and development of tumors. Recently, scholars have gradually valued its important role in liver cancer. This article briefly reviews the structure of Numb molecule, relationship between Numb and tumorigenesis, the molecular mechanism of Numb-regulated tumors, and the role of Numb in the development of liver cancer.

[Ion Chromatography Method of Four Carboxylic Acids in Workplace Air].

Objective: To assess psychological acceptance and occupational stress of medical staff, analyze the relationship among personality, psychological acceptance and occupational stress and discuss the direct or indirect effects of personality to occupational stress. Methods: The gaseous four kinds of carboxylic acids in the workplace air were simultaneously collected by silica gel tube, and then desorbed by deionized water and eluted by ion chromatograph. The the content was detected by conductivity detector. Results: The linear relationship was good in the concentration range of about 0~140 mg/L. The correlation coefficient r>0.999, and the maximum detection limit was 4.5 µg/mL. The sampling efficiency of the four carboxylic acids ranges from 96.10%~100.27%. Through the sample added recovery experiments, the low and high content of the silicone tube samples were detected; and the range of desorption efficiency was 82.18%~100.12%; the range of precision was 0.70%~3.71%. Conclusion: This method adopts deionized water to desorb samples, and the application of ion chromatography detection have reached the requirements of《Guidelines for the establishment of occupational health standards, Part 4: Determination of chemical substances in workplace air》, which can be used in four kinds of workplace air detection of carboxylic acid compounds.

[A concentration-response observation of hydromorphone combined with ropivacaine in labor analgesia].

Objective: To investigate the median effective dose (ED(50)) of hydromorphone and the appropriate concentration of ropivacaine combined with hydromorphone in epidural labor analgesia. Methods: One hundred and forty nulliparous women undergoing labor selected for delivery with epidural analgesia were enrolled in our hospital from January to June 2016. The first of top 50 women received 0.12% ropivacaine plus 20 µg/ml hydromorphone complex solution, then sequential women were used the modified sequential method to determine the ED(50) and ED(95) of hydromorphone. The other 90 women were randomly divided and receieved 0.08% ropivacaine and 15 µg/ml hydromorphone(H1 group), 0.10% ropivacaine and 15 µg/ml hydromorphone (H2 group), 0.12% ropivacaine and 15 µg/ml hydromorphone (group H3) respectively for epidural labor analgesia. In the course of labor, block levels of epidural analgesia, the Bromage scores, analgesia scores and fetal heart rate-uterine concraction were monitored. In addition, onset time of anesthesia, labor time, mode of delivery, cases of increased oxytocin using, neonatal Apgar score, incidence of nausea and vomiting, itching and fetal heart reduction were recorded. Results: The ED(50) and ED(95) values of hydromorphone were 10.49 (95% CI: 8.89-11.79) and 15.15 (95% CI: 13.25-22.25) µg/ml respectively. The onset time in group H1 was significantly longer than those in group H2 and H3((14.23±3.82) , ( 11.32±2.16), (10.83±2.56)min, respectively), the difference was statistically significant (t=5.854, 6.212, all P<0.05). Analgesic VAS score at 30, 60 and 90 min time points in H1 group was significantly higher than that in H2 group and H3 group (all P<0.05). VAS score at withdrawal in H1 group was significantly higher than that in group H3 ( (3.25±0.75) vs (0.27±0.12) ), the difference was statistically significant ( t=9.314, P<0.05). VAS scores at the fourth, fifth, sixth contractions after analgesia in H1 group were significantly higher than those in H2 and H3 groups (all P<0.05). The incidence of motor nerve block in group H3 was higher than that in group H1 and group H2 (26.67%, 6.66%, 3.33%, respectively), the difference was statistically significant (χ(2)=6.413, 4.320, all P<0.05). Conclusions: 0.10% ropivacaine combined with 15 µg/ml hydromorphone has a good analgesic effect, slight motor block, high safety and worthy clinical application for labor analgesia.

Reduced expression of serum miR-204 predicts poor prognosis of gastric cancer.

Deregulation of microRNAs (miRNAs) is implicated in the initiation and progression of gastric cancer (GC). Previous studies have demonstrated that miR-204 was downregulated in GC tissues. However, its expression profile in serum samples and its potential for clinical value remain unknown. Real-time PCR was performed to evaluate the expression level of serum miR-204 in patients with GC. The association between serum miR-204 expression level and the clinical outcome of GC was then investigated. Our results showed that the expression of miR-204 in serum samples from GC patients was significantly lower than that in the healthy controls (P < 0.01). Serum miR-204 expression level of GC patients was significantly upregulated after receiving surgical resection (P < 0.01). In addition, serum miR-204 was associated with lymph node metastasis (P = 0.016), tumor differentiation (P = 0.001), and TNM stage (P = 0.005). GC patients with low serum miR-204 expression had shorter overall survival than those with high serum miR-204 expression (P = 0.004). Multivariate analysis revealed that serum miR-204 expression level was an independent risk factor for this malignant disease (HR = 3.629, 95%CI = 2.828-8.146, P = 0.015). In conclusion, our findings indicate that serum miR-204 may be employed as a novel biomarker for monitoring the treatment response and predicting the prognosis of GC.

Step-flow kinetics model for the vapor-solid-solid Si nanowires growth.

Vapor-solid-solid (VSS) process has recently received continued attention as an alternative to grow Si nanowire. In comparison with common vapor-liquid-solid (VLS) growth with liquid catalyst, VSS growth can prevent the catalyst species from incorporating into nanowires with deep-level impurity, and achieve the compositionally abrupt interfaces by restraining the so-called "reservoir effect". However, despite the huge advances in experimental observations with in situ electron microscopy, VSS growth still remains much less understood in theory. Here, we developed a general mass-transport-limited kinetic model to describe the VSS growth process of Si nanowires by considering three surface diffusion processes and a slow interface diffusion process, where the former determines the atoms supplies way, while the latter dominates the growth of nanowires. The present model is not only well consistent with the available experimental data of Si nanowire, but also gives a clear physical image for the successive side-to-side ledge flow VSS growth.

[Determination of 1-hydroxy-2-butanone in urine by gas chromatography].

Objective: To establish a method for the determination of 1-hydroxy-2-butanone in urine by gas chromatography. Methods: Urine samples were acidified with hydrochloric acid, and then stored in the refrigerator . After thawing under natural conditions, urine volume of 2.0 ml was extracted by C18 solid phase, using methanol volume of 2.0 ml elution, detected by gas chromatography with FID detector and quantified by external standard method. Results: The measurement range of method is 0.986 ~32.88 µg/ml, correlation coefficient (r) =0.999 7, detection limit is 0.13 µg/ml; The recovery is 88.1% ~92.2% ; The precision is 1.4% ~3.2%. Conclusion: This method has a high sensitivity and simple processing. All technical indicators can meet the requirements of the determination method.

Impact of adherence to GOLD guidelines on 6-minute walk distance, MRC dyspnea scale score, lung function decline, quality of life, and quality-adjusted life years in a Shanghai suburb.

We performed a 1-year cluster-randomized field trial to assess the effect of standardized management of chronic obstructive pulmonary disease (COPD) on lung function and quality of life (QOL) measures in patients in China. We used the Global Initiative for Chronic Obstructive Lung Disease (GOLD) treatment guidelines and assessed indexes including pulmonary function, QOL, quality-adjusted life years (QALY), Medical Research Council (MRC) dyspnea scale, 6-min walk distance (6-MWD), number of emergency visits, and frequency of hospitalization. Of a total of 711 patients with chronic cough and asthma, 132 were diagnosed as having COPD and 102 participated in this study [intervention group (N = 47); control group (N = 55)]. We found that adherence to GOLD guidelines had a perceivable impact on 6-MWD, MRC dyspnea scale score, and QOL. The average QALY increased by 1.42/person/year in the intervention group, but declined by 0.95/person/year in the control group. We conclude that standardized management improves disease severity, QOL, and QALY in COPD patients when treatment protocols adhere to GOLD guidelines.

First Report of Phomopsis longicolla Causing Leaf Spot on Soybean in China.

During July 2012, leaf spots affecting 60% of the leaves were observed on soybean cultivar He Feng 60 in fields near Shenyang City, Liaoning Province, leading to 5 to 10% yield loss. The leaf spots were associated with the leaf margins and were irregularly shaped, with brown to black margins and surrounded by a thin, yellow halo. Often, several spots merged to form large necrotic areas, which contained numerous pycnidia on the underside of the leaf. Small pieces (5 mm2) were excised from the margin of diseased and healthy tissue, surface-sterilized in 70% ethanol solution for 30 s and 0.1% mercuric chloride solution for 1 min, washed in three changes of sterile distilled water, and transferred to plates containing potato dextrose agar (PDA). Cultures were maintained in an incubator at 25°C with a 12 h dark/light photoperiod for 5 to 7 days. On PDA, colonies were white with yellow areas, floccose, dense, and moderately fast growing, attaining a diameter of 3.9 mm after 5 days and 9.0 mm after 14 days. Finally, large black stromata appeared after 28 days at 25°C. The conidiomata pycnidia were black, stomatic, globose, length 83.6 to 232 µm, width 37.9 to 146.3 µm and produced α-conidia that were unicellular, hyaline, sometimes two-guttulate, length 4.75 to 8.25 µm, width 1.50 to 3.00 µm. ß-Conidia were not observed. To confirm the morphological identification, the ribosomal internal transcribed spacers (ITS1-5.8S-ITS2) from isolates were sequenced (GenBank Accession No. KC460334). The PCR products were cloned into a pMD-19T Cloning Vector (Sangon Biotech, Shanghai, China). The clones were purified with TIANprep Mini Plasmid Kit (Tiangen Biotech, Beijing, China) to get the full-length ITS sequence. BLAST analysis of the isolates showed 100% nucleotide sequence identity with Phomopsis longicolla (AY745021). Four additional primer pairs-large subunit (NL1/NL4), beta-tublin gene (Bt2a/Bt2b), translation elongation factor 1α gene(EF1-728F/EF1-986R), and act gene(ACT-512F/ACT-783R) (1,2)-were amplified and sequenced as described above. The large subunit gene, ß-tubulin gene, and translation elongation factor 1α gene from isolates were sequenced (Sangon Biotech). BLAST analysis indicated that the isolates had 100% nucleotide sequence identity with P. longicolla (AB107259, HQ333514, and AF398896). Because the act gene sequence of P. longicolla was not in the NCBI database, this sequence had 94% nucleotide sequence identity with P. cuppatea (JN230389). To fulfill Koch's postulates, five leaves on five healthy soybean plants were inoculated with a conidial suspension (106/ml). Plants inoculated with sterile water served as the noninoculated controls. Plants were incubated in the greenhouse at 25°C. All the inoculated leaves developed pinhead spots on the leaves, gradually increasing to large brown spots. Spots were irregularly shaped, brown and necrotic in the center, and surrounded by a yellow halo. Black pycnidia appeared after 10 days, whereas the noninoculated control plants remained asymptomatic. P. longicolla was consistently recovered from all inoculated plants, except the control. Morphological description of isolates was similar to that of Hobbs (3). However, as described by Hobbs and others, P. longicolla conidiomata pycnidia have prominent necks more than 200 µm long, opening by apical ostioles; locules are uniostiolate or multiostiolate, globose, up to 500 µm wide. The pycnidia size of isolates by frozen section method was smaller than that of Hobbs. Based on morphological and sequence comparisons, the pathogen of leaf spot disease is caused by P. longicolla. This is the first reported leaf spot caused by P. longicolla on soybean in China. References: (1) T. Boekhou et al. Stud. Mycol. 38:75, 1995. (2) P. W. Crous et al. Stud. Mycol. 75:37, 2013. (3) T. W. Hobbs et al. Mycologia 77:535, 1985.