ObjectiveTo construct traditional Chinese medicine （TCM） diagnostic scale of turbid toxin syndrome in order to provide corresponding reference for the standardization of TCM syndromes and studies. MethodsWe systematically searched the Chinese Medical Dictionary （CMD）， China Knowledge Network （CNKI）， Wanfang Data Knowledge Service Platform （WF） and VIP database for TCM classics and modern literature on turbid toxin syndrome， and initially screened the four diagnosis information of turbid toxin syndrome， established a pool of information entries， and conducted a cross-sectional clinical survey. Discrete trend method， correlation coefficient method， Cronbach's coefficient method， and factor analysis method were applied to objectively screen the entries. The diagnostic scale of turbid toxin syndrome were constructed through three rounds of Delphi method expert survey to determine the scale entries， using hierarchical analysis to get the judgement matrix scores and relative weight of each entry， after passing consistency test and then isometric expansion of the relative weight of the entries to get the weight of each entry and assign the value. ResultsA total of 35 articles were included， 45 entries were obtained after the initial screening. After the clinical investigation， 12 entries were not suitable by the discrete trend method， 23 entries not suitable by correlation coefficient method， 13 entries by the internal consistency screening were removed with the Cronbach's alpha coefficient rising， and 10 entries not suitable by the factor analysis method. Twenty-two entries were retained after objective screening by the combined use of the four statistical methods. The positive coefficients of experts in the three rounds of Delphi method of expert consultation were 96.67%， the coefficients of expert authority were 0.834， 0.856， and 0.867， and the coefficients of co-ordination were 0.126， 0.326， and 0.312， respectively. After consulting with clinical experts， and three rounds of Delphi method survey and hierarchical analysis method weight assignment， the diagnostic scale entries of turbid toxin syndrome were finally established. Primary symptoms： dark red or purple and dusky tongue， yellowish greasy or dry coating （10 points）； sticky and unpleasant stools （8 points）； disharmony of tastes including halitosis， sticky and greasy taste in the mouth， dry mouth and bitter taste in the mouth （6 points）； unfavourable or yellowish or red urination （5 points）； and dark complexion （4 points）. Secondary symptoms： heavy body （3 points）； dizziness （3 points）； profuse， sticky， foul-smelling secretions （2 points）； wiry and slippery， or slippery， or slippery and rapid pulse （2 points）； feeling of hardness in the abdomen （1 point）. ConclusionUsing Delphi method combined with the hierarchical analysis method， combining qualitative and quantitative study， a diagnostic scale of turbid toxin syndrome was initially developed.

Objective:Novel research and development institutions are crucial strategic forces for technological innovation in the new era. This article analyzed the problems faced by novel research and development institutions in medicine field during their construction and operation, and proposed solutions and countermeasure suggestions, aiming to provide theoretical and practical guidance for their future development.Methods:The author summarized the construction experience of similar institutions domestic and international through literature research and multiple case studies, and explored and analyzed the operation and management mechanisms of novel research and development institutions based on the practice of Xiang An Biomedicine Laboratory.Results:The construction of novel research and development institutions in medicine field faced various problems, such as the existence of blind spots in policy support, limited attractiveness to high-level talents, weak self-renewal capabilities, and inadequate incentive and evaluation systems. It was necessary to establish a management model and operation system that were different from those of traditional research and development institutions.Conclusions:Novel research and development institutions in the medical field should fully leverage their systemic advantages to establish efficient management and incentive systems, build a suitable talent pool, and strengthen the transformation of scientific and technological achievements. Government departments at all levels should provide a comprehensive package of policy support, offer stable financial investment during the initial construction phase, and design appropriate evaluation systems.

WS9326A is a peptide antibiotic containing a highly unusual N-methyl-E-2-3-dehydrotyrosine (NMet-Dht) residue that is incorporated during peptide assembly on a non-ribosomal peptide synthetase (NRPS). The cytochrome P450 encoded by sas16 (P450_Sas) has been shown to be essential for the formation of the alkene moiety in NMet-Dht, but the timing and mechanism of the P450_Sas-mediated α,β-dehydrogenation of Dht remained unclear. Here, we show that the substrate of P450_Sas is the NRPS-associated peptidyl carrier protein (PCP)-bound dipeptide intermediate (Z)-2-pent-1'-enyl-cinnamoyl-Thr-N-Me-Tyr. We demonstrate that P450_Sas-mediated incorporation of the double bond follows N-methylation of the Tyr by the N-methyl transferase domain found within the NRPS, and further that P450_Sas appears to be specific for substrates containing the (Z)-2-pent-1'-enyl-cinnamoyl group. A crystal structure of P450_Sas reveals differences between P450_Sas and other P450s involved in the modification of NRPS-associated substrates, including the substitution of the canonical active site alcohol residue with a phenylalanine (F250), which in turn is critical to P450_Sas activity and WS9326A biosynthesis. Together, our results suggest that P450_Sas catalyses the direct dehydrogenation of the NRPS-bound dipeptide substrate, thus expanding the repertoire of P450 enzymes that can be used to produce biologically active peptides.

Information technology has become an important driver to facilitate higher education developments in the context of new medical sciences. A new “virtual-real combination” experimental teaching model was designed and created through integrating information technology with experimental teaching by Experimental Teaching Center of Basic Medical Sciences and Department of Pathogen Biology, Nanjing Medical University and was applied in Human Parasitology teaching, which achieved satisfactory teaching effectiveness. This new model showed effective to deepen the understanding of the basic human parasitology knowledge, improve the operative skills, and cultivate the moral literacy and comprehensive capability among medical students. This report presents the teaching protocols and implementation, teaching effectiveness and evaluation, and experiences of comprehensive schistosome experiments.

Objective: To explore the status of sugar sweetened beverages(SSBs) and its associated factors among primary and middle school students in Shenzhen, providing reference for nutrition and health education. Methods: A random number table and convenience sampling method was used to select 40 135 primary and middle school students aged 6-18 years in Shenzhen. Data was collected to investigate their SSBs knowledge and associated factors. Results: The proportion of SSBs knowledge score less than 60 points, between 60-79 points and 80-100 points were 5.6%, 41.9% and 52.4%. Multivariate analysis showed that age(4-6 grade, junior middle and high school), gender(female), parents education(high school or vocational schools, colleges and universities as bachelors, masters or doctors), students pay attention to the ingredient list (seldom, none), parental dissuasion or reward behavior (only forced dissuasion, giving no dissuasion, occasional reward, no reward), and the storing beverages at home (seldom, none) were associated with total SSBs knowledge score and milk containing beverage knowledge score ( β =-0.79，-1.19，-1.74，0.58， 1.20 ，1.81，2.98，3.13，2.70，4.85，6.34,6.41，-0.99，-0.78，-1.81，-2.40，5.85，6.26，0.61，1.92， P <0.05). Age(4-6 grade, junior middle and high school), gender(female), father s education background(with colleges and universities as bachelor, masters or doctors), mother s education background(high schools or vocational schools, colleges and universities as bachelors, masters or doctors), parents dissuation behaviors(giving no dissuation), parents rewarding behavior(seldom, none), storing beverages at home(seldom, none) were associated with the total SSBs knowledge score and milk containing beverage knowledge score( β =-0.68， -0.92 ，-0.49，0.26，0.51，1.05，1.09，0.90，1.93，2.62，2.55，-0.68，0.93，1.13，0.21，0.92， P <0.05). Conclusion Primary and middle school students have moderate to high level of SSBs knowledge. It is necessary for students and their parents to learn more SSBs related nutrition healthy knowledge, and to reduce home availability of SSBs.

Background Long-term excessive consuming sugar-sweetened beverages have a negative impact on health. In order to decrease the consumption of sugar-sweetened beverages and create a healthy food environment, the Health Commission of Shenzhen Municipality pioneered to enforce health warning labels presented in commercial locations vending sugar-sweetened beverages based on relevant provisions of the Health Regulations of Shenzhen Special Economic Zone，but its effect has not yet been evaluated. Objective To evaluate the impact of presenting health warning labels in commercial locations vending sugar-sweetened beverages in Shenzhen. Methods A multi-stage stratified sampling method was used to randomly select one street in each of the 10 districts (excluding the Shenzhen Shantou Special Cooperation Zone) of Shenzhen followed by a convenience sampling to select sampling sites to conduct an undercover investigation on the presentation of health warning labels for sugar-sweetened beverages in six different types of venues (n=232) such as shopping malls, ordinary supermarkets/convenience stores, self-service vending machines, catering service places, medical institutions, and venues serving minors' education and activities. At the same time, 238 site managers, 1002 adult consumers, and 7396 child and adolescent consumers were interviewed. Results Among 213 commercial locations vending sugar-sweetened beverages, the rate of health warning label installation was 26.3%, with the highest installation rate in shopping malls (55.0%). Among site managers, 47.8% were aware that commercial locations were required to install health warning labels, and 50.0% were aware of the standards for setting up health warning labels. The higher the awareness of relevant regulations, the higher the rate of installation of health warning labels. More than half of site managers (55.3%) believed that after installing health warning labels, the sales of sugar-sweetened beverages and sugar-sweetened beverages in large-volume packages had decreased compared to the same period in previous years. Most of the interviewed consumers indicated that if they saw the health warning labels for sugar-sweetened beverages, they would buy less, give up purchasing, or choose low-sugar or sugar-free beverages, and also discourage their family members or friends from drinking such beverages. Compared with participants without awareness of the health warning labels, both adult and child and adolescent consumers with awareness of the health warning labels believed that the installation is beneficial to their good eating habits and reported a higher proportion of discouraging family members or friends from drinking such beverages, with a lower frequency of consuming sugar-sweetened beverages. Conclusion Health warning labels for sugar-sweetened beverages have a significant effect on promoting behavior changes, and both site managers and citizens have a high level of support for them. However, in view of the low voluntary compliance rate of commercial locations and the installation rate of health warning labels, the publicity and enforcement of the Health Regulations of Shenzhen Special Economic Zone should be enhanced.

Objective:To prepare camel-derived nanoantibodies that can bind to the recombinant protein VirB12 antigen with high affinity and lay the foundation for further research.Methods:Xinjiang Bactrian camels were immunized six times with VirB12 recombinant protein, total RNA was extracted from lymphocytes isolated from peripheral blood, and the VHH gene fragment was amplified by nested PCR to construct a phage VHH display library. ELISA solid-phase affinity and enrichment methods were used for screening. After three rounds of affinity screening, the clones enriched in the second and third rounds were randomly picked out, and the binding of a nanoantibody with soluble expression to VirB12 was analyzed by ELISA. After sequence determination and multiple alignment, repetitive sequences were removed, and finally five non-redundant sequences were obtained, which were named D1, E6, H8, H9, and H10. The five identified nanoantibody genes were transformed into the WK6 strain, and the soluble expression of an intercellular substance was carried out at 16 °C. After purified expression of Ni-NTA, the binding ability and thermal stability of nanoantibodies and the antigen VirB12 protein were detected by Western Blot and ELISA.Results:Five strains of nanoantibodies were expressed in WK6 bacteria in soluble form. SDS-PAGE showed that the purity of five anti-VirB12 nanoantibodies was close to 90%, and they had high antigen-binding activity and obvious antigen-antibody concentration dependence. All four strains of nanoantibodies showed high thermal stability, and after being treated at 90 ℃, they could still retain more than 60% binding activity.Conclusions:In the study, a VHH phage display library with a capacity of 2.8×10 8 cfu/ml was constructed from Xinjiang Bactrian camel lymphocytes immunized with VirB12 recombinant protein. Five anti-VirB12 nanoantibodies with high affinity and thermal stability were obtained through solid-phase screening and enrichment and soluble monoclonal ELISA detection. These results laid the foundation for further development of VirB12 nanoantibodies.

Age-related macular degeneration (AMD) is a multifactorial disease affected by environmental factors and genetic variation, which is a major cause of irreversible vision loss in the elderly. miRNA is a kind of endogenous non-coding RNA, which plays an important role in the pathogenesis of AMD, such as oxidative stress, pathological neovascularization and inflammation, by inhibiting or silencing the expression of transcription genes. miRNA has unique advantages in terms of ease synthesis, targeting and additive effect, a large number of experiments have proved the therapeutic potential of miRNA in AMD, which is expected to become a new method for the treatment of AMD in the future. Since the pathogenesis of AMD has not been fully elucidated, it is still necessary to continue to study the pathogenesis of AMD, the biological effects and mechanisms of various miRNA in the occurrence and development of AMD, and observe its therapeutic effects in AMD, so as to provide more effective options for the precise prevention and treatment of AMD.

Objective:To determine the content of triptolide in different parts of Tripterygii radix by high performance liquid chromatography. Methods:Tripterygii radix was determined by high performance liquid chromatography with Agilent Technologies C18 column (4.6 mm × 250 mm, 5 μm), acetonitrile-water (33:67) as mobile phase, flow rate of 1 ml/min, the column temperature of 30 ℃, injection volume of 20 ml and wavelength of 218 nm. Results:The linear relationship of triptolide was good in the range of 0.204-2.040 μg ( r=0.999 9), and the average recovery rate was 93.35%; RSD was 1.56% ( n=6). The lignin content in different parts of root was higher than that of the skin. Conclusions:The method is simple, rapid and accurate, which could be used to determine the content of triptolide in Tripterygii radix.

Objective To assess the correlation between serum bilirubin levels (including direct bilirubin and indirect bilirubin) and estimated glomerular filtration rate (eGFR), urinary albumin excretion in type 2 diabetic mellitus patients. Methods A total of 2 390 patients with type 2 diabetes were included. Serum total bilirubin, direct bilirubin, indirect bilirubin, eGFR, urine albumin/uric creatinine rate (UACR) and other information were collected retrospectively. Patients were divided into 4 groups based on the total bilirubin, direct bilirubin and indirect bilirubin separately (from low to high T1 to T4 group, D1 to D4 group and I1 to I4 group). Results UACR was significantly higher in T1 group than that in T2, T3 and T4 groups: 18.79 (10.67, 53.99) mg/g vs. 15.91 (9.86, 36.12), 16.10 (9.35, 29.75) and 16.38 (9.95, 33.44) mg/g (P＜0.01); and eGFR was significantly lower. In D1 to D4 group and I1 to I4 groups, there were statistical differences in UACR and eGFR (P＜0.01). Pearson correlation analyses showed that serum total bilirubin, direct bilirubin and indirect bilirubin were positively correlated with eGFR (r＝0.107, 0.076 and 0.110; P＜0.01), and inversely correlated with UACR (r＝-0.093,-0.078 and-0.090; P ＜ 0.01). Multiple linear regression analysis showed linear relationships between serum total bilirubin, direct bilirubin, indirect bilirubin and eGFR, UACR still existed after adjustment of age, gender, duration of diabetes, BMI, glycosylated hemoglobin A1c and diastolic blood pressure (P＜0.05). Conclusions Serum total bilirubin, direct bilirubin and indirect bilirubin are all positively correlated with eGFR, and inversely correlated with UACR.