The ketogenic diet(KD) treatment program planned for a 2 years old and 5 months boy suffered from severe cerebral complications after ependymoma operation in the Children′s Hospital of Nanjing Medical University at December 2018 was analyzed retrospectively.The patient developed various complications, including cerebral hemorrhage, hydrocephalus and hernia after the operation of ependymoma, with coma, fever and convulsion, and 1 month of anti-infection and antiepileptic treatment was ineffective.After 2 weeks of 4∶1 KD, fever and convulsion were effectively relieved without obvious side effects.One month later, the patient regained consciousness, demonstrating that KD can be applied to treat severe cerebral complications after ependymoma operation.

In vitro evolution methods are often used to modify protein with improved characteristics. We developed a directed evolution protocol to enhance the thermostability of the beta-1,3-1,4-glucanase. The thermostability of the enzyme was significantly improved after two rounds of directed evolution. Three variants with higher thermostability were obtained. The mutant enzymes were further analyzed by their melting temperature, halftime and kinetic parameters. Comparing to intact enzyme, the T50 of mutant enzymes 2-JF-01, 2-JF-02 and 2-JF-03 were increased by 2.2 degrees C, 5.5 degrees C and 3.5 degrees C, respectively, the halftime (t1/2, 60 degrees C) of mutant enzymes 2-JF-01, 2-JF-02 and 2-JF-03 were shortened by 4,13 and 17 min, respectively, the V(max) of mutant enzymes were decreased by 8.3%, 2.6% and 10.6%, respectively, while K(m) of mutant enzymes were nearly unchanged. Sequence analysis revealed seven single amino acid mutant happened among three mutant enzymes, such as 2-JF-01 (N36S, G213R), 2-JF-02 (C86R, S115I, N150G) and 2-JF-03 (E156V, K105R). Homology-modeling showed that five of seven substituted amino acids were located on the surface of or in hole of protein. 42.8% of substituted amino acids were arginine, which indicated that arginine may play a role in the improvement of the thermostability of the beta-1,3-1,4-glucanase.This study provide some intresting results of the structural basis of the thermostability of beta-1,3-1,4-glucanase,and provide some new point of view in modifying enzyme for future industrial use.
Amino Acid Sequence ; Bacillus ; enzymology ; Directed Molecular Evolution ; Endo-1,3(4)-beta-Glucanase ; biosynthesis ; chemistry ; genetics ; Enzyme Stability ; Hot Temperature ; Molecular Sequence Data ; Recombinant Fusion Proteins ; biosynthesis ; chemistry ; genetics

BACKGROUNDTo investigate the diagnostic value of chest CT scan combined with telomerase activity and p16 gene methylation from exfoliated cells of sputum in 55 cases of solitary pulmonary nodules (SPN; ≤30 mm)suspected early peripheral lung cancer.

METHODSThe sputum specimens from 34 cases of cancer nodules and 21 cases of benign lesion were detected for telomerase activity by TRAP-PCR-ELISA and p16 gene methylation by PCR-based methylation analysis.

RESULTSThe qualitative diagnostic accuracy of CT scan was 61.8%(34/55) for SPN provided by pathology. Cytology analysis of sputum was positive in 13 cases (38.2%). Telomerase activity was positive in 29 cases: sensitivity was 79.4%, specificity was 90.5%, accuracy was 83.6%; p16 gene methylation was found in 11 cases: sensitivity was 32.4%, specificity was 100.0%, and accuracy was 58.2%. The sensitivity was increased to 86.1% by combination of telomerase activity and p16 gene methylation. Compared with nodules without malignant CT signs, expression of telomerase activity and p16 methylation of SPN with malignant CT signs (lobulation or spiculate protuberance or spicule sign) had a significant difference (P < 0.01).

CONCLUSIONSThe results suggest that chest CT scan combined with telomerase activity and p16 gene methylation detection in sputum for patients with peripheral lung cancer may enhance the diagnostic value of radiology and conventional cytology.

Objective To investigate the patency rate and restenosis after percutaneous transluminal angioplasty (PTA) for the treatment of arteriovenous fistula (AVF) and arteriovenous graft (AVG) stenosis in dialysis patients. Methods The patients who were successfully treated by PTA for the first time in the blood purification center of the 2nd Affiliated Hospital of Nanjing Medical University from January 2016 to June 2017, including 71 cases of AVF in the forearm, 52 cases of AVF in the upper arm and 59 cases of AVG were recorded. The data of different stenosis parts were analyzed before and after treatment and followed up for 12 months. The initial patency rate and assisted-PTA patency rate were observed at 3 months, 6 months, 9 months, and 12 months after ultrasound interventional therapy, and the initial patency time for patients who needed to reintervention among all types of pathways were recorded. Results The initial patency rates at 3 months, 6 months, 9 months and 12 months after ultrasound interventional therapy were 98.59％, 90.14％, 71.93％, 54.93％ respectively in forearm AVF, 90.38％, 65.38％, 42.31％, 32.69％ respectively in upper arm AVF, 91.53％, 32.20％, 6.78％, 1.69％ respectively in AVG, and the PTA-assisted patency rates were 98.59％, 97.18％, 95.77％, 94.37％ respectively in forearm AVF, 92.31％, 86.54％, 84.62％, 80.77％respectively in upper arm AVF, 100.00％, 98.31％, 96.61％, 93.22％ respectively in AVG, while the initial patency time was (8.99 ± 3.54) months in forearm AVF, (6.33 ± 3.01) months in upper arm AVF, (4.80 ± 1.40) months in AVG respectively. Conclusions Ultrasound can comprehensively evaluate the function of peripheral vascular access, guide PTA treatment, and evaluate treatment outcomes. Ultrasound intervention therapy has best initial patency rate for forearm AVF stenosis. The prognosis of upper arm AVF stenosis PTA is relatively poor due to the easy cephalic stenosis. Although AVG has a short interval of restenosis, it can achieve a better long-term patency rate through regular intervention with ultrasound intervention.

Objective:To perform cross-cultural adaption of the KING′s Parkinson′s Disease Pain Scale (KPPS), explore its reliability and validity in Chinese Parkinson′s disease (PD) patients, and to create the new version of the pain scale which adapts to the Chinese PD patients.Methods:This study enrolled 225 patients, including 121 men and 104 women who were selected from the Outpatient Center of Movement Disorders Clinic of the Second Affiliated Hospital of Soochow University from July 2018 to July 2020. All patients completed the evaluation of the Chinese Version of KPPS (KPPS-CV). According to the preliminary evaluation results, the expert group modified KPPS-CV after discussion, and developed a Modified KPPS-CV (MKPPS-CV). These patients then completed the MKPPS-CV evaluation during the 3-month follow-up. Cross-cultural adaptation was performed according to published international guidelines that include translation, back-translation, expert review, and pretesting. The following psychometric properties were evaluated: basic item analysis; floor and ceiling effects; construct validity; content validity; criterion validity (Spearman′s rho between the KPPS-CV and Numeric Rating Scale); internal consistency reliability (Cronbach′s alpha); test-retest reliability (intra-class correlation coefficient, ICC).Results:In item analysis, 50% of the items had poor discrimination (critical ratio<3.0), and floor effect was found in all domains (proportion of 0 point>15%). The items were reclassified after exploratory factor analysis. The content validity of item 3, item 10 and item 11 was low (item-level content validity index<0.78). Criterion validity showed the highest correlations (Spearman′s rho>0.88) between the KPPS-CV and Numeric Rating Scale. While overall scale reliability was minimally acceptable at 0.46, which showed a poor reliability of this scale. Test-retest reliability was excellent for each item (Spearman's rho>0.85). The Cronbach′s alpha of MKPPS-CV (0.76) was higher than that of KPPS-CV (0.46). It showed a great improvement after the modifying.Conclusions:When using scales that are not developed for local populations, differences in culture and clinical practices should be taken into account. MKPPS-CV is an acceptable, valid measure to evaluate pain in Chinese PD patients, which is more suitable for Chinese people.

OBJECTIVE: Identifying novel strategies to prevent particulate matter (PM)-induced lung injury is crucial for the reduction of the morbidity of chronic respiratory diseases. The combined intervention represented by herbal formulae for simultaneously targeting multiple pathological processes can provide a more beneficial effect than the single intervention. The aim of this paper is therefore to design a safe and effective medicinal and edible Chinese herbs (MECHs) formula against PM-induced lung injury. METHODS: PM-induced oxidative stress, inflammatory response and apoptosis A549 cell model were used to screen anti-oxidant, anti-inflammatory and anti-apoptotic MECHs, respectively. A network pharmacology method was utilized to rationally design a novel herbal formula. Ultra performance liquid chromatography-mass spectrometer was utilized to assess the quality control of MECHs formula. The excretion of magnetic iron oxide nanospheres of the MECHs formula was estimated in zebrafish. The MECH formula against PM-induced lung injury was investigated with mice experiments. RESULTS: Five selected herbs were rationally designed to form a new MECH formula, including Citri Exocarpium Rubrum (Juhong), Lablab Semen Album (Baibiandou), Atractylodis Macrocephalae Rhizoma (Baizhu), Mori Folium (Sangye) and Polygonati Odorati Rhizoma (Yuzhu). The formula effectively promoted the magnetic iron oxide nanospheres excretion in zebrafish. The mid/high dose formula significantly prevented PM-induced lung damage in mice by enhancing the activity of SOD and GSH-Px, reducing the MDA and ROS level and attenuating the upregulation of pro-inflammatory cytokine (IL-6, IL-8, IL-1β and TNF-α), down regulating the protein expression of NF-κB, STAT3 and Caspase-3. CONCLUSION Our findings suggest that the effective MECHs formula will become a novel strategy for preventing PM-induced lung injury and provide a paradigm for the development of functional foods using MECHs.