Objective: To extract and purify total flavonoids from Periploca forrestii Schltr.(P. forrestii),and test the in vitro inhibitory activity of the total flavonoids and two flvaonoidal compounds in P. forrestii,so as to provide a reference for studies on the related medicinal substances in P. forrestii. Methods: Total flavonoids were extracted from P. forrestii and then purified by the column chromatography on macroporous resin and polyamide columns. The content of total flavonoids was determined according to the Lambert-Beer’s law. The in vitro xanthine oxidase(XOD)inhibitory ac- tivity was assayed for total flavonoids and the two flavonoidal compounds by the ultraviolet spectrophotometry. Results The purified total flavonoids had a content of more than 95%. The total flavonoids and two flavonoidal compounds all showed an inhibitory effect on XOD in vitro,with the inhibitory rate enhanced with increasing concentration. The IC50 of the total flavonoids as well as the two flavonoidal compounds,quercetin-3-O-α-L-pyranoside(QP)and quercetin-7-O-β- D-glucopyranoside(QG)were 608.9,221.2 and 261.2 μg/ml,respectively. Conclusion: The total flavonoids as well as the two flvaonoidal compounds QP and QG in P. forrestii all showed the in vitro inhibitory activity on XOD.

OBJECTIVETo better comprehend the molecular structure and physiological function of the housefly larval peritrophic matrix (PM), a mass spectrometry approach was used to investigate the PM protein composition.

METHODSThe PM was dissected from the midgut of the third instar larvae, and protein extracted from the PM was evaluated using SDS-PAGE. A 1D-PAGE lane containing all protein bands was cut from top to bottom, the proteins in-gel trypsinised and analysed via shotgun liquid chromatography- tandem mass spectrometry (LC-MS/MS).

RESULTSIn total, 374 proteins, with molecular weights varying from 8.225 kD to 996.065 kD and isoelectric points ranging from 3.83 to 11.24 were successfully identified, most identified proteins were mainly related to immunity, digestion, nutrient metabolism and PM structure. Furthermore, many of these proteins were functionally associated with pattern binding, polysaccharide binding, structural constituent of peritrophic membrane and chitin binding, according to Gene Ontology annotation.

CONCLUSIONThe PM protein composition, which provides a basis for further functional investigations of the identified proteins, will be useful for understanding the housefly larval gut immune system and may help to identify potential targets and exploit new bioinsecticides.

Animals ; Chitin ; metabolism ; Gastrointestinal Tract ; metabolism ; Houseflies ; metabolism ; Insect Proteins ; metabolism ; Larva ; metabolism ; Proteomics

ObjectiveTo compare the clinical effects of Shang Ring scissor circumcision (SC) and electrotome circumcision (EC) in the treatment of redundant prepuce or phimosis.Methods: Results: Conclusion.

METHODSThis retrospective study included 524 patients with redundant prepuce or phimosis, 422 treated by SC and 120 by EC. We made comparisons between the two groups of patients in the operation time, intra- and post-operative pain scores, pain scores before, at and after ring removal, wound healing time, and incidence rates of postoperative edema and incision dehiscence.

RESULTSThe operation time was longer in the SC than in the EC group (［59.99±5.39］ vs ［39.94±4.94］ sec, P<0.05), but there were no significant differences between the two groups in the intraoperative pain scores (1.02±0.74 vs 1.08±0.59, P>0.05) or the pain scores within 24 h after operation (6.74±1.01 vs 6.56±1.06, P>0.05), 24 h prior to ring removal (1.14±0.69 vs 1.10±0.64, P>0.05), and after ring removal (2.73±0.74 vs 2.85±0.75, P>0.05) except at ring removal, which was remarkably lower in the SC than in the EC group (3.56±0.47 vs 4.77±0.58, P<0.05). The wound healing time was markedly shorter in the former than in the latter (［14.11±1.26］ vs ［39.78±7.55］ d, P<0.05), but the incidence rate of incision dehiscence showed no significant difference between the two groups (4.03% ［17/422］ vs 9.17% ［11/120］, P>0.05). The rate of postoperative satisfaction with the external penile appearance was 100% in both of the two groups.

CONCLUSIONSShang Ring scissor circumcision is preferred to electrotome circumcision for its advantages of less pain at ring removal and shorter healing time despite its longer operation time.

Aim: To investigate the effects of alpha lipoic acid (ALA) on Notch2, TLR4, NLRP3 and inflammatory factor expression in renal tissues of diabetes mellitus(DM) rats, and to explore its possible mechanism of anti-inflammatory and anti-fibrosis by Alpha lipoic acid. Methods: The diabetic rat model was established by streptozotocin. The rats were divided into two groups; the DM group and ALA group. Meanwhile, and another eight rats were used as normal control (NC group). After eight weeks, the rats were sacrificed to detect the relevant biochemical parameters and oxidative stress indexes. In addition, immunohistochemical staining was employed to detect the protein expression localization sites of Notch2, TLR4 and NLRP3. Western blot analysis was used to detect the expression of Notch2, TLR4, NLRP3 and collagen FV proteins in renal tissues. ELISA was utilized to detect the inflammatory factor expression of IL-6 and TNF-α. Results: Compared with NC group, the levels of blood glucose, 24h urine protein, total cholesterol and triglyceride all increased in DM group, and the activity of T-AOC was reduced whereas MDA content was up-regulated in DM group, all items but blood glucose were significantly reduced in ALA group, and the activity of T-AOC remarkably increased, while MDA content was reduced in ALA group. Compared with NC group, the protein levels of Notch2, TLR4, NLRP3 and collagen IV in kidneys were raised, and the inflammatory factor expression of IL-6 and TNF-α significantly increased in DM group. Conclusions: ALA may down-regulate the inflammatory signal of TLR4 and NLRP3 in kidney of diabetic rats, and reduce the expression of inflammatory factor and the accumulation of extracellular matrix via inhibiting the expression of Notch2 at protein level.

OBJECTIVE: To study the transport of lipoamide (LAM) and lipoic acid (LA) in Caco-2 cell monolayer model in vitro. METHODS: Effects of LAM and LA on the survival rate of Caco-2 cells were investigated by MTS, the bi-directional transport of lipoamide and lipoic acid from the intestinal cavity side (apical side, AP) to the basal side (basolateral side,BL) was investigated. The cumulative transport volume, apparent permeability coefficient (Papp) and transport percentage were calculated,and the relationships between transport volume and concentration and time were further studied. RESULTS: The transport amounts of LAM and LA were increased in time-and concentration-dependent manners, the Papps of LAM and LA (AP→BL) were 2.443 44×10-5-2.392 91×10-5 and 8.179 78×10-6-7.897 25×10-6 cm•s-1, and the Papps(BL→AP) were 2.258 13×10-5-2.214 3×10-5 and 8.267 98×10-6-7.926 73×10-6 cm•s-1, respectively. CONCLUSION: In the transport test of Caco-2 cells, LAM is superior to LA, suggesting that it is well absorbed orally and has high bioavailability. But it is still necessary to verify the pharmacokinetic data in vivo.

Objective: To investigate the correlation between the level of P34H expression and the activity of hyaluronidase in human spermatozoa. Methods: Eighty eight semen samples were collected, 68 cases were in the infertile group, and 20 in the normal control group. Semen routine analysis was referred to the WHO standard method. According to the difference of semen parameters, 68 cases of infertile males were divided into the infertile group with normal semen parameters and the abnormal semen parameters. Western blotting was used to detect the level of P34H expression on spermatozoa. The P34H-positive rate on human spermatozoa was determined by indirect immunofluorescent staining using anti-P34H antibody. The HYD-positive rate and HYD-activity intensity in all samples were examined by improved fixedsubstrate film method. Results: The level of P34H protein expression and the percentage of the P34H-positive rate in infertile groups with normal semen parameters and abnormal semen parameters were significantly lower than that in the control group (P<0.05). The activity of HYD (HYD-positive rate, HYD-activity intensity) in the infertile groups with normal semen parameters and abnormal semen parameters were also significantly lower than that in control group (P < 0.05). The relation between the P34H protein expression and HYD-positive rate, HYD-activity intensity had a significant positive correlation (r = 0. 449, 0. 431; P < 0.01); the relation between the the percentage of P34H-positive rate and HYD-positive rate, HYD-activity intensity had a significant positive correlation (r=0.727, 0.691;P <0.01). Conclusion: The level of P34H protein expression and the percentage of the P34H-positive rate are decreased, while the activity of HYD (HYD-positive rate, HYD-activity intensity) is reduced in male infertility.

Background Chronic excessive exposure to fluoride can cause damage to the central nervous system and a certain degree of learning and memory impairment. However, the associated mechanism is not yet clear and further exploration is needed. Objective Using 4D unlabelled quantitative proteomics techniques to explore differentially expressed proteins and their potential mechanisms of action in chronic excessive fluoride exposure induced brain injury. Methods Twenty-four SPF-grade adult SD rats, half male and half male, were selected and divided into a control group and a fluoride group by random number table method, with 12 rats in each group. Among them, the control group drank tap water (fluorine content<1 mg·L⁻¹), the fluoride group drank sodium fluoride solution (fluorine content 10 mg·L⁻¹), and both groups were fed with ordinary mouse feed (fluoride content<0.6 mg·kg⁻¹). After 180 d of feeding, the SD rats were weighed, and then part of the brain tissue was sampled for pathological examination by hematoxylin-eosin (HE) staining and Nissl staining. The rest of the brain tissue was frozen and stored at −80 ℃. Three brain tissue samples from each group were randomly selected for proteomics detection. Differentially expressed proteins were screened and subcellular localization analysis was performed, followed by Gene Ontology (GO) function analysis, Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis, cluster analysis, and protein-protein interaction analysis. Finally, Western blotting was used to detect the expression levels of key proteins extracted from the brain tissue samples. Results After 180 d of feeding, the average weight of the rats in the fluoride group was significantly lower than that in the control group (P<0.05). The brain tissue stained with HE showed no significant morphological changes in the cerebral cortex of the fluoride treated rats, and neuron loss, irregular arrangement of neurons, eosinophilic changes, and cell body pyknosis were observed in the hippocampus. The Nissl staining results showed that the staining of neurons in the cerebral cortex and hippocampus of rats exposed to fluoride decreased (Nissl bodies decreased). The proteomics results showed that a total of 6927 proteins were identified. After screening, 206 differentially expressed proteins were obtained between the control group and the fluoride group, including 96 up-regulated proteins and 110 down-regulated proteins. The differential proteins were mainly located in cytoplasm (30.6%), nucleus (27.2%), mitochondria (13.6%), plasma membrane (13.6%), and extracellular domain (11.7%). The GO analysis results showed that differentially expressed proteins mainly participated in biological processes such as iron ion transport, regulation of dopamine neuron differentiation, and negative regulation of respiratory burst in inflammatory response, exercised molecular functions such as ferrous binding, iron oxidase activity, and cytokine activity, and were located in the smooth endoplasmic reticulum membrane, fixed components of the membrane, chloride channel complexes, and other cellular components. The KEGG significantly enriched pathways included biosynthesis of secondary metabolites, carbon metabolism, and microbial metabolism in diverse environments. The results of differential protein-protein interaction analysis showed that the highest connectivity was found in glucose-6-phosphate isomerase (Gpi). The expression level of Gpi in the brain tissue of the rats in the fluoride group was lower than that in the control group by Western blotting (P<0.05). Conclusion Multiple differentially expressed proteins are present in the brain tissue of rats with chronic fluorosis, and their functions are related to biosynthesis of secondary metabolites, carbon metabolism, and microbial metabolism in diverse environments; Gpi may be involved in cerebral neurological damage caused by chronic overdose fluoride exposure.

OBJECTIVE: To explore the genetic basis of a child with holoprosencephaly. METHODS: Genomic DNA of the child was extracted and subjected to whole exome sequencing. Suspected variant was verified by Sanger sequencing of her family members. RESULTS: Cranial MRI suggested lobulated holoprosencephaly with partial absence of corpus callosum. Genetic testing revealed that she has carried a heterozygous c.517C>G (p.His173Asp) variant of the SIX3 gene, for which both of her parents were of wild type. Based on the American College of Medical Genetics and Genomics guidelines, the c.517C>G variant of SIX3 gene was predicted to be pathogenic (PS2+PM1+PM2+PM5+PP3). CONCLUSION The SIX3 gene c.517C>G variant probably underlay the multiple malformations in this child. Above finding has enabled her definite diagnosis.
Child ; Family ; Female ; Heterozygote ; Holoprosencephaly/genetics* ; Humans ; Mutation ; Whole Exome Sequencing

Objective To investigate the effect of whole body vibration training on biomechanics and Wnt3a protein expression of the femur. Methods Forty-eight female SD rats were randomly divided into sham operation group, osteoporosis group and whole body vibration group, 16 in each group. The bone morphometric parameters were measured by Micro-CT, mechanical parameters of bone structure and materials were measured by three-point bending test, protein expression of Wnt3a and β-catenin was measured by Western blotting, and gene expression of Wnt3a, β-catenin, cyclin D1 and tcf1 was detected by qRT-PCR. ResultsCompared with sham operation group, bone mineral density (BMD), bone volume fraction (BVF), trabecular number, trabecular thickness and cortical bone thickness in osteoporosis group were decreased, and trabecular space was increased; compared with osteoporosis group, BMD, BVF, trabecular number, trabecular thickness and cortical bone thickness in whole body vibration group were increased, and trabecular space was decreased. Compared with sham operation group, the maximum load, elastic load and deflection of osteoporosis group were significantly reduced; compared with osteoporosis group, the maximum load, elastic load and deflection of whole body vibration group were significantly increased. Compared with sham operation group, the maximum stress, elastic stress, maximum strain and elastic modulus in osteoporosis group decreased significantly; compared with the osteoporosis group, the elastic stress, maximum strain and elastic modulus in whole body vibration group increased significantly. Compared with sham operation group, Wnt3a, β-catenin protein and gene expression decreased, cyclin D1, tcf1 gene expression also decreased; compared with osteoporosis group, Wnt3a, β-catenin protein and gene expression increased, cyclin D1, tcf1 gene expression increased as well. Conclusions Whole body vibration training can improve biomechanical properties of the femur and expression of Wnt3a protein in osteoporotic rats. The research findings provide laboratory reference data for the prevention and treatment of osteoporosis by whole body vibration training.

Air Pollution ; China ; Humans ; Meteorological Concepts ; Outpatients ; Particulate Matter/analysis* ; Skin Diseases/epidemiology*