Heterologous Expression and Characterization of a pH-Stable Chitinase from Micromonospora aurantiaca with a Potential Application in Chitin Degradation.

To promote the bioconversion of marine chitin waste into value-added products, we expressed a novel pH-stable Micromonospora aurantiaca-derived chitinase, MaChi1, in Escherichia coli and subsequently purified, characterized, and evaluated it for its chitin-converting capacity. Our results indicated that MaChi1 is of the glycoside hydrolase (GH) family 18 with a molecular weight of approximately 57 kDa, consisting of a GH18 catalytic domain and a cellulose-binding domain. We recorded its optimal activity at pH 5.0 and 55 °C. It exhibited excellent stability in a wide pH range of 3.0-10.0. Mg2+ (5 mM), and dithiothreitol (10 mM) significantly promoted MaChi1 activity. MaChi1 exhibited broad substrate specificity and hydrolyzed chitin, chitosan, cellulose, soluble starch, and N-acetyl chitooligosaccharides with polymerization degrees ranging from three to six. Moreover, MaChi1 exhibited an endo-type cleavage pattern, and it could efficiently convert colloidal chitin into N-acetyl-D-glucosamine (GlcNAc) and (GlcNAc)2 with yields of 227.2 and 505.9 mg/g chitin, respectively. Its high chitin-degrading capacity and exceptional pH tolerance makes it a promising tool with potential applications in chitin waste treatment and bioactive oligosaccharide production.

[Introduction of pertussis surveillance system and evaluation of its operational effect in Tianjin].

The pertussis surveillance system has been established since 2009 in Tianjin, and continuously improved over the past 10 years. This system determines the definition and classification of pertussis, establishes simple and feasible sampling methods and laboratory detection methods in clinical practice, standardizes the report management of pertussis cases and the treatment of epidemic situations. After the implementation of the surveillance system, the number of reported pertussis cases increased from 26 in 2009 to 802 in 2017, the number of diagnosed cases increased from 19 in 2009 to 662 in 2017, the reported incidence rate of pertussis increased from 0.16/100 000 in 2009 to 4.28/100 000 in 2017, and the number of medical institutions of reporting perutssis cases increased from 2 in 2009 to 53 in 2017. The specimen collection rate of the reported cases reached up to 93.66%. These results show that the sensitivity of pertussis surveillance has been improved and show that the data from the surveillance system may reflect more precisely the epidemical characteristics of perutssis in Tianjin.

Impact of the number of examined lymph nodes on outcomes in patients with lymph node-negative gallbladder carcinoma.

AIM: To determine whether the number of examined lymph nodes (LNs) is correlated with the overall survival of gallbladder carcinoma (GBC) patients. METHODS: Patients were collected from the Surveillance Epidemiology and End Results database (2004-2013) and categorized by the number of LNs into six groups: 1 LN, 2 LNs, 3 LNs, 4 LNs, 5 LNs, and ≥ 6 LNs. Survival curves for overall survival were plotted with a Kaplan-Meier analysis. The log-rank test was used for univariate comparisons. RESULTS: In a cohort of 893 patients, the median number of examined LNs was two for the entire cohort. The survival for the 1 LN group was significantly poorer than those of the stage I and II disease groups and for the entire cohort. By dichotomizing the number of LNs from 1 to 6, we found that the minimum number of LNs that should be examined was four for stage I, four or five for stage II, and six for stage IIIA disease. Therefore, for the entire cohort, the number of examined LNs should be at least six, which is exactly consistent with the American Joint Committee on Cancer criteria. CONCLUSION: The examination of higher numbers of LNs is associated with improved survival after resection surgery for N0 GBC. The guidelines for GBC surgery, which recommend that six LNs be examined at least, are statistically valid and should be applied in clinical practice widely.

Stage III should be subclassified into Stage IIIA and IIIB in the American Joint Committee on Cancer (8th Edition) staging system for pancreatic cancer.

AIM: To ascertain the prognostic role of the T4 and N2 category in stage III pancreatic cancer according to the 8th edition of the American Joint Committee on Cancer (AJCC) classification. METHODS: Patients were collected from the Surveillance Epidemiology and End Results (SEER) database (2004-2013) and were divided into three groups: T(1-3)N2, T4N(0-1), and T4N2. Overall survival (OS) and disease-specific survival (DSS) of patients were evaluated by the Kaplan-Meier method. RESULTS: For the first time, we found a significant difference in OS and DSS between T(1-3)N2/T4N(0-1) and T4N2 but not between T(1-3)N2 and T4N(0-1). A higher grading correlated with a worse prognosis in the T(1-3)N2 and T4N2 groups. CONCLUSION: Patients with stage T4N2 had a worse prognosis than those with stage T(1-3)N2/T4N(0-1) in the 8th edition AJCC staging system for pancreatic cancer. We recommend that stage III should be subclassified into stage IIIA [T(1-3)N2/T4N(0-1)] and stage IIIB (T4N2).

Effects and action mechanisms of sodium fluoride (NaF) on the growth and cephalotaxine production of Cephalotaxus mannii suspension cells.

To explore the effects of NaF on the growth and cephalotaxine production of Cephalotaxus mannii suspension cells, NaF was added into the C mannii cell suspension cultures at day 15 of culture time. It is documented that NaF suppressed cell growth but enhanced cephalotaxine production. The largest yield obtained of cephalotaxine reached to 9.57mg/L when the cultures were treated with an appropriate dosage of 15mg/L NaF, which was 3.7 times that of the control cultures (2.58mg/L). Additionally, NaF markedly enhanced the activity of glucose 6-phosphate dehydrogenase (G6PDH) and reduced the level of hydrogen peroxide (H2O2) of cells. It was also found that NaF weakened the oxidative damage of cell membrane and led to lower content of malonyl dialdehyde (MDA) in NaF-treated cells compared with the control cells. The MDA content of NaF-treated cells decreased 91% compared to the controls. Although lower membrane lipid peroxidation in NaF-treated cells, its membrane permeability was higher than the control cells and showed a high product secret rate. What is more, NaF boosted the activity of phenylalanine ammonium-lyse (PAL), but did not burst a peak of PAL activity in the time curve of PAL activity. These results indicated NaF acted as an inhibitor of the Enbden Parnas (EMP) pathway, not as an elicitor to promote cephalotaxine production.

[Compensation-fitting extraction of dynamic spectrum based on least squares method].

Extraction method for dynamic spectrum (DS) with a high signal to noise ratio is a key to achieving high-precision noninvasive detection of blood components. In order to further improve the accuracy and speed of DS extraction, linear similarity between photoelectric plethysmographys (PPG) at each two different wavelengths was analyzed in principle, and an experimental verification was conducted. Based on this property, the method of compensation-fitting extraction was proposed. Firstly, the baseline of PPG at each wavelength is estimated and compensated using single-period sampling average, which would remove the effect of baseline drift caused by motion artifact. Secondly, the slope of least squares fitting between each single-wavelength PPG and full-wavelength averaged PPG is acquired to construct DS, which would significantly suppress random noise. Contrast experiments were conducted on 25 samples in NIR wave band and Vis wave band respectively. Flatness and processing time of DS using compensation-fitting extraction were compared with that using single-trial estimation. In NIR band, the average variance using compensation-fitting estimation was 69.0% of that using single-trial estimation, and in Vis band it was 57.4%, which shows that the flatness of DS is steadily improved. In NIR band, the data processing time using compensation-fitting extraction could be reduced to 10% of that using single-trial estimation, and in Vis band it was 20%, which shows that the time for data processing is significantly reduced. Experimental results show that, compared with single-trial estimation method, dynamic spectrum compensation-fitting extraction could steadily improve the signal to noise ratio of DS, significantly improve estimation quality, reduce data processing time, and simplify procedure. Therefore, this new method is expected to promote the development of noninvasive blood components measurement.

[Noninvasive measurement of serum total protein content by near-infrared reflection spectra with tongue inspection].

The technology of tongue near-infrared reflectance spectra was used for human serum total protein (TP) content of noninvasive testing for the first time. Reflectance spectrum on the tongue tips of 58 volunteers was collected, and the biochemical values of serum total protein were recorded at the same time. The samples were separated into two parts: training set and prediction set. Two prediction models were established using PCA combined with BP neural network and PLS. Using PCA-BP model to predict the prediction set, the average relative error is 7.35%, RMSEP was 6.3771 g x L(-1), and the correlation coefficient was 0.9021. Using PLS model to predict the prediction set, the average relative error is 4.77%, RMSEP was 0.1304 g x L(-1), and the correlation coefficient was 0.9718. It was approved that reflectance spectra of tongue can be used to predict TP accurately and noninvasively.

[D-value estimation of dynamic spectrum based on the statistical methods].

To realize the noninvasive concentration detection of blood components and improve the drawbacks existing in the time-domain single-trial estimation method of "dynamic spectrum" (DS), the D-value estimation method based on the statistical properties was proposed. We extracted the absolute difference between two corresponding values of each wavelength to make up the DS, selected the valid DSs from the DSs of different times by statistic method, and the valid DSs were superimposed and averaged as the final output of the DS. Data collected from 48 volunteers were processed by the D-value estimation and the single-trial estimation, respectively; and then the comparison was carried out between the two methods. Compared with the single-trial estimation, the valid DSs extracted by the D-value estimation were slightly better in denoising; And the average number of the remained valid DSs is improved from 48 to 130; the average of mean square error among the valid DSs is improved from 0.39 to 0.006; the speed of data processing is increased by nearly 20 times. The new method can significantly improve the quality of the extraction of DS.

Syntheses and characterization of novel oxoisoaporphine derivatives as dual inhibitors for cholinesterases and amyloid beta aggregation.

A series of 3-substituted (5c-5f, 6c-6f) and 4-substituted (10a-10g) oxoisoaporphine derivatives were synthesized. It was found that all these synthetic compounds had IC50 values at micro or nano molar range for cholinesterase inhibition, and most of them could inhibit amyloid ß (Aß) self-induced aggregation with inhibition ratio from 31.8% to 57.6%. The structure-activity relationship studies revealed that the derivatives with higher selectivity on AChE also showed better inhibition on Aß self-induced aggregation. The results from cell toxicity study indicated that most quaternary methiodide salts had higher IC50 values than the corresponding non-quaternary compounds. This study provided potentially important information for further development of oxoisoaporphine derivatives as lead compounds for the treatment of Alzheimer's disease.

Paclitaxel production using co-culture of Taxus suspension cells and paclitaxel-producing endophytic fungi in a co-bioreactor.

The co-culture of the suspension cells of Taxus chinensis var. mairei and its endophytic fungi, Fusarium mairei, in a 20-L co-bioreactor was successfully established for paclitaxel production. The co-bioreactor consists of two-unit tanks (10 L each) with a repairable separate membrane in the center, culturing Taxus suspension cells in one tank and growing fungi in another. By optimizing the co-culture conditions, there was a desirable yield of paclitaxel in Taxus cell cultures. The Taxus cell cultures by co-culture produced 25.63 mg/L of paclitaxel within 15 days; it was equivalent to a productivity of 1.71 mg/L per day and 38-fold higher than that by uncoupled culture (0.68 mg/L within 15 days). The optimum conditions for co-culture in the co-bioreactor were: B5 medium, inoculating fungi when Taxus cells had grown for 5 days in the co-bioreactor, hydrophilic separate membrane in the center of the co-bioreactor, and air flow rate of 1:0.85 v/v/m in fungus cultures.

Interactions of Taxol-producing endophytic fungus with its host (Taxus spp.) during Taxol accumulation.

Endophytic fungi (Fusarium mairei) culture broth (EFCB) was added to cell suspension cultures of Taxus cuspidata. After 5 days, cultures of T. cuspidata given 4 ml of EFCB produced a maximal yield of 6.11 mg/l paclitaxel, with a release ratio of 75%, 2- and 6.8-fold, respectively, greater than the controls. The active element in EFCB is an exopolysaccharide of approximately 79 kD. Endophytic fungi produced 0.19 mg/l of paclitaxel in its producing medium. However, when the supernatant of Taxus cell suspension cultures from day 20 was added to the paclitaxel-producing medium, the biomass of fungi decreased by 24% and the yield of paclitaxel by 45%. In a co-culture system of plant and fungus, the yield of paclitaxel (12.8 mg/l) was >2-fold higher than that in the EFCB-treatment system.

[Effects of occupation on health of traffic policemen in a city].

OBJECTIVE: To investigate the health and work status of traffic policemen in a city, and analyze the effect of the occupation on health of them. METHODS: The prevalent investigation was used to investigate the health status of on-the-job traffic policemen and the effect of the occupation on the health of traffic policemen was studied. RESULTS: Among 4486 traffic policemen, 1604 working outside. The average time of working outside was (12.0 +/- 7.9) years and the average time of standing was (7.0 +/- 1.7) hours/day. The incidence of naso-pharyngitis, arthropathy, photosensitive dermatitis, heat stroke and chilblain, noise-induced deafness in the outside-traffic policemen were higher than that of the policemen working indoors. Eye diseases were higher in those who worked outside for 20-25 years. The varicosis of the lower extremity was higher in those who stood outside over 4 hours every day. The average period of onset of disease was 7-8 years. CONCLUSION: The occupational stress of outside-traffic policemen was bigger in the city. The environmental pollution and the bad condition are the risky factors of 8 kinds of diseases among traffic policemen. The average period of onset of the disease is 7-8 years.

[Evaluation on the role of hepatitis A vaccine in the prevention and control of hepatitis A in Tianjin city].

OBJECTIVE: To evaluate the effects of prevention and control strategies on hepatitis A. METHODS: Surveillance data on hepatitis A from 1990 to 2006 in Tianjin was analyzed, and the coverage rate of hepatitis A vaccine among targeted population was estimated, to compare the anti-HAV IgG level of children younger than 15 years old in 1999 and in 2005. RESULTS: Results showed that a) the morbidity of hepatitis A decreased from 25.26/10(5) in 1990 to 0.82/10(5) in 2006; b) the ratio of hepatitis A in viral hepatitis decreased from 30.43% in 1990 to 1.05% in 2006; c) the estimated coverage rate was 72.7%; d) the positive rate of anti-HAV among children younger than 15 years old in 2005 was distinctly higher than that in 1999. CONCLUSION: Positive results showed that it was successful to use hepatitis A vaccine as the strategy to prevent and control hepatitis A in the past five years in Tianjin.