[Clinical comprehensive evaluation of Jinsang Liyan Pills/Capsules in treatment of chronic pharyngitis].

According to the Guidelines for clinical comprehensive evaluation of Chinese patent medicine(2022 version), this study comprehensively compared the clinical value of Jinsang Liyan Pills/Capsules with that of another commonly used Chinese patent medicine(drug A).(1)Effectiveness: Jinsang Liyan Pills/Capsules had antimicrobial, anti-inflammatory, and pain-relieving effects and can improve the total response rate in the treatment of chronic pharyngitis. Moreover, they took effect faster than the control group.(2)Safety: Jinsang Liyan Pills/Capsules did not cause acute toxicity and long-term toxicity, with low incidence of adverse reactions, which were mild and alleviated after drug withdrawal. Therefore, the risk of Jinsang Liyan Pills/Capsules was under control.(3)Economy: Jinsang Liyan Pills/Capsules had lower cost per course of treatment than drug A. The incremental cost-effectiveness ratio(ICER) of Jinsang Liyan Pills combined with Jinsang Qingyin Pills was-39.97 yuan compared with conventional treatment. The ICER of Jinsang Liyan Pills compared with amoxicilin was 0.01 yuan. The results meant that Jinsang Liyan Pills/Capsules had a cost-effectiveness advantage.(4)Innovation: Jinsang Liyan Pills/Capsules had reasonably formula and wide indications, meeting the clinical needs. Moreover, they had been authorized four patents of advanced manufacturing technology.(5)Suitability: the storage and administration of Jinsang Liyan Pills/Capsules were convenient, with clear instruction of medication.(6) Accessibility: Jinsang Liyan Pills/Capsules had sufficient drug reserve, caused low economic burden of patients, and presented environmental bearing capacity. Finally, Jinsang Liyan Pills/Capsules were scored 79.10 points, and drug A 67.93 points. The experts reached the consensus of grade A for Jinsang Liyan Pills/Capsules, which can be directly converted into decision making. The result of this comprehensive evaluation of Jinsang Liyan Pills/Capsules highlight the clinical advantages in the treatment of chronic pharyngitis and lay a foundation for the standardized research on the clinical basic research of the drug in the future.

[Clinical comprehensive evaluation of Jinsang Sanjie Pills/Capsules in treatment of vocal nodule/polyp of vocal cord].

This study used health technology assessment methods and multi-criteria decision analysis(MCDA) model, according to the guideline for clinical comprehensive evaluation of Chinese patent medicine, we developed this assessment tool. The comprehensive evaluation score of Jinsang Sanjie Pills/Capsules is calculated based on the additive model. This score is calculated by "quantitative evaluation software v1.0 for clinical comprehensive evaluation of Chinese patent medicines" which developed by the project team. The evaluation yielded the following results.(1)Effectiveness: compared with the control group, Jinsang Sanjie Pills/Capsules can improve the total effectiveness rate of vocal nodule/polyp of vocal cord, and improve the symptoms and signs.(2)Safety: Jinsang Sanjie Pills/Capsules did not show acute toxicity and long-term toxicity. The most common adverse reaction was gastrointestinal system damage, all of the adverse reactions were either improved or cured.(3)Economy: from the perspective of the health system, evaluating the single use or combination of Jinsang Sanjie Pills/Capsules with conventional medication in the treatment of vocal nodule/polyp of vocal cord is relatively effective and cost-effective compared to conventional medication, with a stable cost-effectiveness advantage.(4) Innovation: Jinsang Sanjie Pills/Capsules are used for the treatment of slow throat paralysis(vocal nodules, polyp of vocal cord, thickening of vocal mucosa) caused by heat toxin accumulation, Qi stagnation and blood stasis, and the resulting hoarseness. Jinsang Sanjie Pills/Capsules have good innovation and targeted indications.(5) Suitability: the investigated doctors, pharmacists and patients all believed that Jinsang Sanjie Pills/Capsules have good suitability.(6)Accessibility: Jinsang Sanjie Pills/Capsules are included in the category B of the National Basic Medical Insurance, Work Injury Insurance, and Maternity Insurance Drug Catalogue(2021 edition), which have good cost-effectiveness and affordability for medical insurance and self-paid patients. Jinsang Sanjie Pills/Capsules do not contain endangered animals and plants. The supply of raw materials can meet the demand of production at present. The comprehensive evaluation score is 76.06 points. Based on all dimensions of evidence, 71.4% experts consensus on Jinsang Sanjie Pills/Capsules is class A, which can be directly converted into decision making. This study comprehensively evaluated the clinical application value of Jinsang Sanjie Pills/Capsules in the treatment of vocal nodule/polyp of vocal cord, so as to provide evidence for their rational clinical use and regulatory decision-making.

Distribution of α-Synuclein Aggregation in the Peripheral Tissues.

Parkinson's disease (PD) is a chronic neurodegenerative disease mainly characterized by movement disorders and other non-motor symptoms, including the loss of dopaminergic neurons in the substantia nigra parts. Abnormal α-synuclein aggregation in the brain is closely associated with the loss of dopaminergic neurons. α-synuclein can propagate in the central nervous system (CNS) and periphery under pathological conditions. Many researches have focused on its aggregation and distribution in the CNS and explored its relationship with PD. But in recent years, the distribution of α-synuclein in peripheral tissues have been paid much attention. This review summarized the distribution of α-synuclein in the choroid plexus, blood, saliva, gastrointestine and other tissues, and discussed the potential mechanism of α-synuclein aggregation, providing a basis for the early diagnosis and intervention of PD.

Risk factors for food sensitization in children with atopic dermatitis: a single-center cross-sectional study. / ç‰¹åº”æ€§çš®ç‚Žæ‚£å„¿é£Ÿç‰©è‡´æ•å±é™©å› ç´ åˆ†æžï¼šä¸€é¡¹å•ä¸­å¿ƒæ¨ªæ–­é¢ç ”ç©¶.

OBJECTIVES: To study the risk factors for food sensitization and the influence of food sensitization on quality of life and clinical signs in children with atopic dermatitis (AD). METHODS: A retrospective analysis was performed on the medical data of 241 children with AD, including demographic features, age of onset, severity of AD, quality of life, physical examination results, skin prick test (SPT) results, serum total IgE levels, and eosinophil count. According to the results of SPT, the children were divided into a food sensitization group (n=127) and a non-food sensitization group (n=114). The multivariate logistic regression analysis was used to identify the risk factors for food sensitization in children with AD. RESULTS: The prevalence rate of food sensitization was 52.7% (127/241) in the children with AD. The multivariate logistic regression analysis showed that birth in autumn or winter, age of onset of AD<12 months, severe AD, and total IgE>150 IU/mL were risk factors for food sensitization (P<0.05). Compared with the non-food sensitization group, the food sensitization group had a significantly poorer quality of life (P=0.008) and significantly higher prevalence rates of non-specific hand/foot dermatitis and palmar hyperlinearity (P<0.05). Compared with the single food sensitization group, the multiple food sensitization group had more severe AD and a significantly higher proportion of children with exclusive breastfeeding or total IgE>150 IU/mL (P<0.05). CONCLUSIONS: The AD children born in autumn or winter, or those with early onset (<12 months), severe AD or total IgE>150 IU/mL have a higher risk of food sensitization. The AD children with food sensitization have a poorer quality of life and are more likely to develop non-specific hand/foot dermatitis and palmar hyperlinearity.

Maternal yeast-based nucleotide supplementation decreased stillbirth by regulating nutrient metabolism.

BACKGROUND: As an enzymatic product of yeast, yeast-based nucleotide (YN) is rich in nucleotides. To test the effects of maternal dietary supplementation with YN during late pregnancy on placental nutrient transport and nutrient metabolism in neonatal piglets, 64 pregnant sows (day 85 ± 3) were assigned into two groups: (i) control (CON) and (ii) treatment (YN; 4 g kg-1 ). Blood, placenta and liver samples of neonates during delivery were collected. RESULTS: The results showed that maternal YN supplementation decreased stillbirth rate and intra-uterine growth restriction rate (P < 0.05). In addition, maternal YN supplementation increased total serum protein, albumin and total cholesterol (P < 0.05). Furthermore, in neonatal piglets in the YN group, both serum amino acidand nucleotide profiles were affected, as well as liver amino acid, and fatty acid profiles were regulated (P < 0.05). Moreover, maternal YN supplementation increased liver mRNA expression of SLC28A3, SLC29A1, SLC29A2, PC, PCK1, FBP1, SREBP1c, HSL and CYP7a1 of neonatal piglets (P < 0.05). Meanwhile, there was a decrease in placental gene expression of EAAT2, EAAT3, LAT1 and PAT1, as well as lower protein expression of peroxisome proliferator-activated receptor (PPAR)γ, AKT, phosphorylated-AKT, phosphorylated-mammalian target of rapamycin (mTOR) and Raptor, in the YN group (P < 0.05). CONCLUSION: Taken together, these results indicate that maternal YN supplementation regulates placental nutrient transport by regulating the mTOR complex 1-PPAR pathway, and affects the liver metabolism of nucleotides, amino acids and fatty acids in neonatal piglets, thereby improving the reproductive performance of sow to a certain extent. © 2020 Society of Chemical Industry.

Bioactive flavonoid dimers from Chinese dragon's blood, the red resin of Dracaena cochinchinensis.

Seven flavonoid dimers, biflavocochins A-G, together with six known compounds were isolated from the red resins of Dracaena cochinchinensis (Chinese dragon's blood). Their structures were elucidated based on extensive spectroscopic analysis. The absolute configurations of 1-7 was assigned by experimental and quantum chemical calculated ECD spectra, and that of 4 was further established by X-ray diffraction analysis using Cu Kα radiation. Compounds 1-3 are novel dimers of homoisoflavonoid and dihydrochalcone with a unique dibenzopyran ring. Compounds 2, 6, 7 exhibited moderate PTP1B inhibitory activities in an enzyme assay. Compound 1 showed neuroprotective effect on serum deficiency-induced cellular damage in PC12 cells.

Exploring Genome-wide DNA Methylation Profiles Altered in Kashin-Beck Disease Using Infinium Human Methylation 450 Bead Chips.

To understand how differentially methylated genes (DMGs) might affect the pathogenesis of Kashin-Beck disease (KBD). Genome-wide methylation profiling of whole blood from 12 matched KBD and controls pairs was performed using a high-resolution Infinium 450 K methylation array. In total, 97 CpG sites were differentially methylated in KBD compared to the normal controls; of these sites, 36 sites were significantly hypermethylated (covering 22 genes) and 61 sites were significantly hypomethylated (covering 34 genes). Of these genes, 14 significant pathways were identified, the most significant P value pathway was type I diabetes mellitus pathway and pathways associated with autoimmune diseases and inflammatory diseases were included in this study. Subsequently, 4 CpG sites in HLA-DRB1 were validated using bisulfite sequencing polymerase chain reaction (BSP) in articular cartilage, and the results showed significant differences in the methylation status between KBD and controls, consistent with the results of the high-resolution array. These results suggested that differences in genome-wide DNA methylation exist between KBD and the controls, and the biological pathways support the autoimmune disease and inflammatory disease hypothesis of KBD.

[Effects of Shaoyao Gancao decoction on contents of amino acids and expressions of receptors in brains of spastic paralysis rats].

To explore the effects of Shaoyao Gancao decoction on contents of amino acids and expressions of receptors in the brains of spastic paralysis rats, the spastic paralysis rat models of stroke convalescence were made by line tethering method. Baclofen was used as the control group, and the experiment group received Shaoyao Gancao decoction at 3∶1 proportions. After 3 weeks, the neurobehavioral scores, muscular tension and pain threshold were measured and compared. High performance liquid chromatography (HPLC) was used to detect the contents of GABA, Gly, Glu, Asp in cerebral cortex. The protein expressions of GABA receptors Aα1, B； NMDA receptor NR1, NR2A and NR2B in cerebral cortex were determined by immunohistochemistry assay. The results showed that the Shaoyao Gancao decoction at 3∶1 proportion could improve the spastic paralysis state after stroke, significantly improve neurological symptoms (P<0.01), decrease muscular tension (P<0.01) and improve pain threshold (P<0.05) as compared with model group. Simultaneously, the contents of inhibitory amino acids GABA and Gly were increased significantly (P<0.01), while with a decrease tendency in excitatory amino acids Glu and Asp (with no statistical significance). In addition, it could significantly increase the protein expressions of neurotransmitter GABA receptors Aα1, and B (P<0.05); reduce the expressions of neurotransmitter NMDA receptors NR1, NR2A and NR2B (P<0.05). These results suggested that the Shaoyao Gancao decoction at 3：1 proportion could effectively relieve spasm and pain. The mechanism might be associated with increasing the contents of inhibitory amino acids and increasing the expressions of their receptors in spastic paralysis rats after stroke, which would consequently enhance the signal transduction of inhibitory amino acids. Meanwhile, there was a decrease tendency in excitatory amino acids, although no significant effect was observed, and it could suppress the expressions of excitatory amino acids receptors, thus weaken the excitatory signal transduction. Thereby the neurotoxicity was relieved eventually. These findings indicated that Shaoyao Gancao decoction could regulate the balance of neurotransmitter system to relieve the spasticity, and eventually achieve tendon tonifying and spasm relieving effect.

Changing Grains for the Prevention and Treatment of Kashin-Beck Disease in Children: a Meta-analysis.

To evaluate the efficacy of changing grains on the prevention and treatment of Kashin-Beck Disease (KBD) in children, community-based trials were acquired from seven electronic databases (up to July 2014). As a result, the methodological quality of the six trials that have been included into our analysis was low. The pooled ORs favoring the prevention and treatment effects of changing grains were 0.15 (95% CI: 0.03-0.70) and 2.13 (95% CI: 1.44-3.16) respectively by meta-analysis. Subgroup analysis demonstrated the pooled OR favoring treatment effect of exchanging grains rather than drying grains both compared with endemic grains. The results showed that changing grains had obvious effects on the prevention and treatment of KBD in children. However, the evidences were limited by the potential biases and confounders. Large and well-designed trials are still needed.

[Comparative study on effects of blood enriching on mouse model of blood deficiency syndrome induced by cyclophosphamide of albiflorin, paeoniflorin on levels of GM-CSF, IL-3 and TNF-α].

OBJECTIVE: To compare the effects and mechanism of blood enriching on mouse model of blood deficiency syndrome induced by cyclophosphamide of albiflorin and paeoniflorin. METHOD: Albiflorin and paeoniflorin were determined by using animal models of blood deficiency syndrome induced by cyclophosphamide. The amount of WBC, RBC, HGB, index of thymus gland and spleen, and the changes of GM-CSF, IL-3 and TNF-α in serum were detected after the treatment. RESULT: Compared with the model group, the amount of WBC in the group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were increased obviously (P < 0.01). The amount of RBC in the group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were increased obviously (P < 0.01, P < 0.001), which did not had a significant difference compared with the same dose. The index of thymus gland in the group of 30 mg x kg(-1) albiflorin was superior to the model group (P < 0.01), the difference was significant compared with the same dose of paeoniflorin (P < 0.05). The GM-CSF in serum in all groups of 30 mg x kg(-1) albiflorin, 15 mg x kg(-1) albiflorin, 30 mg x kg(-1) paeoniflorin and 15 mg x kg(-1) paeoniflorin increased obviously (P < 0.01, P < 0.05, P < 0.01, P < 0.05); The IL-3 in serum in both group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin also increased (P < 0.001). The content of TNF-α in group of 30 mg x kg(-1) albiflorin and 30 mg x kg(-1) paeoniflorin were reduced (P < 0.01), which showed the obvious difference compared with the same dose group (P < 0.01). CONCLUSION: Albiflorin had the effect of blood enriching by regulating the immune function, same with the paeoniflorin. The probable mechanism of nourishing blood and liver of Paeoniae Radix Alba was not only the better effect of adjusting the content of TNF-α, but also might act synergistically with paeoniflorin.

Influence of TPU/EVA Phase Morphology Evolution on Supercritical Carbon Dioxide Extrusion Foaming.

Ethylene-vinyl acetate copolymer (EVA) was added at different contents to the thermoplastic polyurethane (TPU) matrix to form a non-compatible blending system, and foaming materials with high pore density were prepared using the supercritical carbon dioxide extrusion method. The influence of the phase morphology and crystal morphology of the TPU/EVA blend on its foaming behavior was studied. The results show that EVA changed the phase morphology and crystal morphology of the blends, leading to the improved melt viscosity and crystallinity of the blend system. At the same time, interfacial nucleation increases the density of cells and decreases the cell thickness and size, which is beneficial for improving the foaming properties of the blends. For the EVA content of 10% (mass fraction), the cell size is small (105.29 µm) and the cell density is the highest (3.74 × 106 cells/cm3). Based on the TPU/EVA phase morphology and crystal morphology, it is found that the sea-island structure of the blend has better foaming properties than the bicontinuous structure.

Successful treatment of epidermolysis bullosa pruriginosa by dupilumab.

Epidermolysis bullosa pruriginosa (EBP) is a rare variant of dystrophic epidermolysis bullosa caused by COL7A1 gene mutation. Intense pruritus and nodular prurigo-like lesions are the main features of the disease. To date, the treatment strategies for this condition are not well established. Recent studies have indicated that type 2 inflammation plays a role in the pathophysiology of EBP, suggesting Th2 cytokines could be potential therapeutic targets. In this prospective case series study, we reported three patients with EBP, diagnosed by clinical manifestations, histopathological evaluations, and genetic sequencing, two of whom were treated with dupilumab for 20 weeks. Results showed that the clinical symptoms, pruritus, and quality of life of the patients were significantly improved, as measured by the Epidermolysis Bullosa Disease Activity and Scarring Index, the Visual Analog Scale, and the Children's Dermatology Life Quality Index. Serum immunoglobulin E levels also fell gradually over the 20-week treatment period. Immunotyping of Th1/2/17 cell subsets in peripheral blood by flow cytometry revealed a higher Th2 but parallel Th1 and Th17 cell subsets in patients compared to healthy controls, and a significant decrease in Th2 and an increase in Th17 cells after dupilumab administration. Of note, after 20 weeks of dupilumab treatment, the expression of type VII collagen in the basement membrane of the skin lesion of the patients significantly increased, which was evidenced by immunofluorescence analysis. No treatment-related adverse events were documented. Taken together, targeting type 2 inflammation with dupilumab may be an effective and safe treatment option for EBP.

The neuroinflammatory role of glucocerebrosidase in Parkinson's disease.

The lysosomal enzyme glucocerebrosidase (GCase), encoded by the GBA1 gene, is a membrane-associated protein catalyzing the cleavage of glucosylceramide (GlcCer) and glucosylsphingosine (GlcSph). Homologous GBA1 mutations cause Gaucher disease (GD) and heterologous mutations cause Parkinson's disease (PD). Importantly, heterologous GBA1 mutations are recognized as the second risk factor of PD. The pathological features of PD are Lewy neurites (LNs) and Lewy bodies (LBs) composed of pathological α-synuclein. Oxidative stress, inflammatory response, autophagic impairment, and α-synuclein accumulation play critical roles in PD pathogenic cascades, but the pathogenesis of PD has not yet been fully elucidated. What's more, PD treatment drugs can only relieve symptoms to a certain extent, but cannot alleviate neurodegenerative progression. Therefore, it's urgent to explore new targets that can alleviate the neurodegenerative process. Deficient GCase can cause lysosomal dysfunction, obstructing the metabolism of α-synuclein. Meanwhile, GCase dysfunction causes accumulation of its substrates, leading to lipid metabolism disorders. Subsequently, astrocytes and microglia are activated, releasing amounts of pro-inflammatory mediators and causing extensive neuroinflammation. All these cascades can induce neuron damage and death, eventually promoting PD pathology. This review aims to summarize these points and the potential of GCase as an original target to provide some ideas for elucidating the pathogenesis of PD.

Novel carrier-mediated membrane-assisted three-phase liquid-liquid extraction coupled with liquid chromatography-mass spectrometry for the determination of eight biogenic amines in foods.

Analysis of biogenic amines (BAs) is of great importance due to their toxicity and usage as indicators of food freshness. In this study, membrane-assisted three-phase liquid-liquid extraction (MA-3pLLE) was proposed to integrate extraction and back-extraction into a single step using a specially designed U-shaped device. Parameters affecting the performance of the extraction method were optimized. Coupled with liquid chromatography-mass spectrometry, the method was successfully applied for the simultaneous determination of eight BAs without derivatization in apple juice, orange juice, red wine, soy sauce and milk granules, with satisfactory recoveries and RSDs. The calibration curve of the method was linear in the range of 1.00~100.00 ng/mL, with a correlation coefficient (r2) ≥ 0.9908. The limits of detection were in the range of 0.03~1.00 ng/mL. MA-3pLLE is efficient, reproducible, and green and has great potential for application in the one-step extraction of analytes from complex matrices.

Effect of Dual-Task Training on Gait and Balance in Stroke Patients: An Updated Meta-analysis.

OBJECTIVE: The main purpose of this meta-analysis was to evaluate the effect of dual-task training on gait and balance improvement in stroke patients. DESIGN: The PubMed, Embase, Cochrane Library, MEDLINE, CINAHL, CNKI, Wan Fang, and VIP databases were searched from inception to January 28, 2021, for randomized controlled trials investigating the effect of dual-task training on gait and balance intervention in stroke patients. RESULTS: A total of 17 studies with 575 stroke patients that compared the efficacy and safety of dual-task training with those of conventional physical therapy or single-task training were included in this meta-analysis. The meta-analysis showed that the data were as follows under the dual-task training: step length (mean difference = 2.7, 95% confidence interval = 1.33 to 4.08, P = 0.0001); cadence (mean difference = 5.06, 95% confidence interval = 3.37 to 6.75, P < 0.00001); stride length (mean difference = 7.34, 95% confidence interval = 5.47 to 9.22, P < 0.00001); 10-meter walk test times (mean difference = -2.36, 95% confidence interval = -3.70 to -1.02), P = 0.0006); Berg Balance Scale (mean difference = 3.8, 95% confidence interval = 0.04 to 7.55, P = 0.05); Fugl-Meyer motor assessment of lower extremities (mean difference = 2.27, 95% confidence interval = -1.04 to 5.59, P = 0.18). CONCLUSIONS: This meta-analysis showed that dual-task training can improve stroke patients' step length, cadence, stride length, and 10-meter walk test. However, possible advantages in improving balance function need further exploration.

Effects of robotic-assisted gait training on motor function and walking ability in children with thoracolumbar incomplete spinal cord injury.

BACKGROUND: Spinal cord injury (SCI) results in neurological dysfunction of the spinal cord below the injury. OBJECTIVE: To explore the immediate and long-term effects of robotic-assisted gait training (RAGT) on the recovery of motor function and walking ability in children with thoracolumbar incomplete SCI. METHODS: Twenty-one children with thoracolumbar incomplete SCI were randomly divided into the experimental (n = 11) and control groups (n = 10). The control group received 60 min of conventional physical therapy, and the experimental group received 30 min of RAGT based on 30 minutes of conventional physical therapy. Changes in walking speed and distance, physiological cost index (PCI), lower extremity motor score (LEMS), SCI walking index and centre-of-pressure (COP) envelope area score were observed in both groups of children before and after eight weeks of training. The primary outcome measures were the 10-metre walk test (10MWT) and six-minute walk distance (6MWD) at preferred and maximal speeds. In addition, several other measures were assessed, such as postural control and balance, lower limb strength and energy expenditure. RESULTS: Compared with control group, the self-selected walk speed (SWS), maximum walking speed (MWS), 6MWD, PCI, LEMS, COP, and Walking Index for Spinal Cord injury II (WISCI II) of experimental group were improved after treatment. The 6MWD, PCI, COP, and WISCI II after eight weeks of treatment were improved in experimental group. All indicators were not identical at three different time points when compared between two groups. Pairwise comparisons in experimental group suggested that the SWS, MWS, 6MWD, PCI, LEMS, COP, and WISCI II after treatment were higher than those before treatment. The 6MWD, LEMS, COP, and WISCI II after treatment were higher than at the one-month follow-up appointment. The SWS, PCI, LEMS, COP, and WISCI II at the eight-week follow-up appointment were improved. CONCLUSION: Robotic-assisted gait training may significantly improve the immediate motor function and walking ability of children with thoracolumbar incomplete SCI.

Identification of chitinases Is-chiA and Is-chiB from Isoptericola jiangsuensis CLG and their characterization.

A 274-bp conserved fragment of chiA (chiA-CF) was amplified from the genomic DNA of Isoptericola jiangsuensis CLG (DSM 21863, CCTCC AB208287) using the specific PCR primers. Based on chiA-CF sequences, a 5233-bp DNA fragment was obtained by self-formed adaptor PCR. DNA sequencing analysis revealed there were two contiguous open reading frames coding for the precursors of Is-chiA [871 amino acids (aa)] and Is-chiB (561 aa) in the 5233-bp DNA fragment. The Is-chiA and Is-chiB exhibited 58% and 62% identity with ArChiA and ArChiB chitinase from Arthrobacter sp. TAD20, respectively. The Is-chiA and Is-chiB genes were cloned into expression vector pET28a (+) and expressed in Escherichia coli BL21 (DE3) with isopropyl-ß-D-thiogalactopyranoside induction. Is-chiA and Is-chiB were 92 kDa and 60 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and showed chitobiosidase and endochitinase activity, respectively. Is-chiA and Is-chiB were purified by Ni-nitrilotriacetic acid affinity chromatography and the characteristics of both Is-chiA and Is-chiB were studied.

A bibenzyl compound 20C protects rats against 6-OHDA-induced damage by regulating adaptive immunity associated molecules.

Parkinson's disease (PD) is a neurodegenerative disease with complicated pathogenesis. A novel bibenzyl compound 2-[4-hydroxy-3-(4-hydroxyphenyl)benzyl]-4-(4-hydroxyphenyl)phenol (20C) has been shown to have some neuroprotective effects, and its mechanism still needs further research. In this study, we used a 6-hydroxydopamine (6-OHDA)-induced PD rat model to evaluate the protective effect of 20C. Our study found that 20C could improve behavioral defects in 6-OHDA-lesion rats, decrease neuroinflammation and protect their DA neurons. It could inhibit the activity of inducible nitric oxide synthase (iNOS) induced by 6-OHDA, and lead to a decrease in the expression of nitrated-α-synuclein. When exposed to AMT-an inhibitor of iNOS, the nitrated-α-synuclein in PC12 decreased, and 20C demonstrated the same function on nitrated-α-synuclein as AMT. Besides, we also found that nitrated-α-synuclein was displayed in microglia. And 20C could decrease the expression of antigen-presenting molecule major histocompatibility complex I (MHC I) in dopamine (DA) neurons and MHC II in microglia induced by 6-OHDA. So, these imply that nitrated-α-synuclein might act as an endogenous antigen activating adaptive immunity, and the neuroprotection of 20C might be associated with inhibiting the activity of iNOS, decreasing the expression of the antigen molecule nitrated-α-synuclein and the antigen presenting molecule MHC. Our results indicated that inhibiting iNOS might be an effective strategy to protect neurons from oxidative stress.

Tet1 Deficiency Leads to Premature Reproductive Aging by Reducing Spermatogonia Stem Cells and Germ Cell Differentiation.

Ten-eleven translocation (Tet) enzymes are involved in DNA demethylation, important in regulating embryo development, stem cell pluripotency and tumorigenesis. Alterations of DNA methylation with age have been shown in various somatic cell types. We investigated whether Tet1 and Tet2 regulate aging. We showed that Tet1-deficient mice undergo a progressive reduction of spermatogonia stem cells and spermatogenesis and thus accelerated infertility with age. Tet1 deficiency decreases 5hmC levels in spermatogonia and downregulates a subset of genes important for cell cycle, germ cell differentiation, meiosis and reproduction, such as Ccna1 and Spo11, resulting in premature reproductive aging. Moreover, Tet1 and 5hmC both regulate signaling pathways key for stem cell development, including Wnt and PI3K-Akt, autophagy and stress response genes. In contrast, effect of Tet2 deficiency on male reproductive aging is minor. Hence, Tet1 maintains spermatogonia stem cells with age, revealing an important role of Tet1 in regulating stem cell aging.