Imaging evaluation of cerebrospinal fluid otorrhea associated with inner ear malformation in children / 中华耳鼻咽喉头颈外科杂志

Objective: To explore the imaging evaluation of cerebrospinal fluid (CSF) otorrhea associated with inner ear malformation (IEM) in children. Methods: The clinical data of 28 children with CSF otorrhea associated with IEM confirmed by surgical exploration in Beijing Children's Hospital, from Nov, 2016 to Jan, 2021, were analyzed retrospectively,including 16 boys and 12 girls, aged from 8-month to 15-year and 8-month old, with a median age of 4-year old. The shapes of stapes were observed during the exploration surgery, and the imaging features of temporal bone high resolution CT(HRCT) and inner ear MRI pre- and post-operation were analyzed. Results: In 28 children with CSF otorrhea, 89.3%(25/28) had stapes footplates defect during exploration. Preoperative CT showed indirect signs such as IEM, tympanic membrane bulging, soft tissue in the tympanum and mastoid cavity. IEM included four kinds: incomplete partition type I (IP-Ⅰ), common cavity (CC), incomplete partition type Ⅱ (IP-Ⅱ), and cochlear aplasia (CA); 100%(28/28) presented with vestibule dilation; 85.7%(24/28) with a defect in the lamina cribrosa of the internal auditory canal. The direct diagnostic sign of CSF otorrrhea could be seen in 73.9%(17/23) pre-operative MRI: two T2-weighted hyperintense signals between vestibule and middle ear cavity were connected by slightly lower or mixed intense T2-weighted signals, and obvious in the coronal-plane; 100%(23/23) hyperintense T2-weighted signals in the tympanum connected with those in the Eustachian tube.In post-operative CT, the soft tissues in the tympanum and mastoid cavity decreased or disappeared as early as one week. In post-operative MRI, the hyperintense T2-weighted signals of tympanum and mastoid decreased or disappeared in 3 days to 1 month,soft tissues tamponade with moderate intense T2-weighted signal were seen in the vestibule in 1-4 months. Conclusions: IP-Ⅰ, CC, IP-Ⅱ and CA with dilated vestibule can lead to CSF otorrhea. Combined with special medical history, T2-weighted signal of inner ear MRI can provide diagnostic basie for most children with IEM and CSF otorrhea.HRCT and MRI of inner ear can also be used to evaluate the effect of surgery.

Mechanism of Marsdenia tenacissima against ovarian cancer based on network pharmacology and experimental verification / 中国中药杂志

The present study aimed to explore the main active components and underlying mechanisms of Marsdenia tenacissima in the treatment of ovarian cancer(OC) through network pharmacology, molecular docking, and in vitro cell experiments. The active components of M. tenacissima were obtained from the literature search, and their potential targets were obtained from SwissTargetPrediction. The OC-related targets were retrieved from Therapeutic Target Database(TTD), Online Mendelian Inheritance in Man(OMIM), GeneCards, and PharmGKB. The common targets of the drug and the disease were screened out by Venn diagram. Cytoscape was used to construct an "active component-target-disease" network, and the core components were screened out according to the node degree. The protein-protein interaction(PPI) network of the common targets was constructed by STRING and Cytoscape, and the core targets were screened out according to the node degree. GO and KEGG enrichment analyses of potential therapeutic targets were carried out with DAVID database. Molecular docking was used to determine the binding activity of some active components to key targets by AutoDock. Finally, the anti-OC activity of M. tenacissima extract was verified based on SKOV3 cells in vitro. The PI3K/AKT signaling pathway was selected for in vitro experimental verification according to the results of GO function and KEGG pathway analyses. Network pharmacology results showed that 39 active components, such as kaempferol, 11α-O-benzoyl-12β-O-acetyltenacigenin B, and drevogenin Q, were screened out, involving 25 core targets such as AKT1, VEGFA, and EGFR, and the PI3K-AKT signaling pathway was the main pathway of target protein enrichment. The results of molecular docking also showed that the top ten core components showed good binding affinity to the top ten core targets. The results of in vitro experiments showed that M. tenacissima extract could significantly inhibit the proliferation of OC cells, induce apoptosis of OC cells through the mitochondrial pathway, and down-regulate the expression of proteins related to the PI3K/AKT signaling pathway. This study shows that M. tenacissima has the characteristics of multi-component, multi-target, and multi-pathway synergistic effect in the treatment of OC, which provides a theoretical basis for in-depth research on the material basis, mechanism, and clinical application.

China experts' consensus on preventive and interceptive orthodontic treatments of malocclusions of children / 华西口腔医学杂志

Malocclusion is one of the three most common oral diseases reported by World Health Organization(WHO). In China, its incidence rate is rising. Malocclusion seriously affects the dental and maxillofacial function, facial appearance and growth development of nearly 260 million children in China, and what is more, it affects their physical and mental health development. Malocclusion occurrence is related to genetic and environmental factors. Early treatment of malocclusion can create a good dental and maxillofacial development environment, correct abnormal growth and control the adverse effects of abnormal genetic factors. It can effectively reduce the prevalence of children's malocclusion and enhance their physical and mental health. This is an urgent need from the economic perspective of our society, so it has great practical and social significance. Experts from the project group "standard diagnose and treatment protocols for early orthodontic intervention of malocclusions of children" which initiated by China National Health Institute of Hospital Administration wrote the "China Experts' Consensus on Preventive and Interceptive Orthodontic Treatments of Malocclusions of Children", which aims to guide and popularize the clinical practice, improve the clinical theory and practice level, and accelerate the disciplinary development of early treatment of children's malocclusion in China. The consensus elaborates the harmfulness of malocclusion and the necessity of early treatment, and brings up the principles and fundamental contents. Based on the law of dental and maxillofacial development, this paper puts forward the guiding suggestions of preventive and interceptive treatments in different stages of dental development ranging from fetus to early permanent dentition. It is a systematic project to promote and standardize the early treatment of malocclusion. Through scientific and comprehensive stratified clinical practice and professional training, the clinical system of early treatment of malocclusion in China will eventually be perfected, so as to comprehensively care for children's dental and maxillofacial health, and improve their oral and physical health in China.

Effects of Vitrification with Self-made Carriers and Slow Programmed Freezing on Ovarian Tissue of Sheep / 中国医学科学院学报

Objective To investigate the effects of self-made carriers on the cryopreservation of ovarian tissue of sheep. Methods Thirty-two ovaries were randomly assigned to fresh group,programmed freezing group,self-made carrier I vitrification group,and self-made carrier Ⅱ vitrification group.The morphology,proliferation,apoptosis,and estrogen level of the ovarian tissue in each group were observed. Results After cryopreservation,the morphology normal rate of the primordial follicles in programmed freezing group,self-made carrier I vitrification group,and self-made carrier Ⅱ vitrification group were 74.2%,72.8%,and 72.3%,respectively,lower than that(83.7%)in the fresh group(χ

In vitro inhibition by 79 herb injectables of nine human cytochrome P450 isoenzymes: a systematic evaluation of herb-drug interactions / 药学学报

Seventy-nine injectable herb extractions have been approved by the Chinese Food and Drug Administration (CFDA) and are frequently administered intravenously for various diseases. Unfortunately, herb-drug interactions are under-investigated and sometimes overlooked in the clinic. In the present investigation the in vitro inhibition of 9 drug metabolizing enzymes including CYP1A, CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP2E1, and CYP3A was assessed using an appropriate probe substrate for each enzyme with human liver microsomes. Metabolite formation was quantified using a validated and sensitive high-performance liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay. The IC50 of each herb extract was estimated using a concentration range from 5% to 0.5%, and the time-dependent inhibition of the nine CYP450 isoenzymes was also determined. Of the 79 approved iv herb injectables, 37 inhibited CYP1A, 24 inhibited CYP2A6, 41 inhibited CYP2B6, 36 inhibited CYP2C8, 31 inhibited CYP2C9, 41 inhibited CYP2C19, 13 inhibited CYP2D6, 25 inhibited CYP2E1, and 42 inhibited CYP3A with 50% or greater inhibition at a test concentration of 5% (v/v). IC50 differences were noted between pre-incubation or co-incubation assays with HLM for 30 min, with the time-dependent inhibitory (TDI) effects were observed with 2 injectables on CYP1A, 5 injectables on CYP2A6, 5 injectables on CYP2B6, 6 injectables on CYP2C8, 1 injectable on CYP2D6 and 6 injectables on CYP3A. Collectively, the results demonstrate that potential herb-drug interactions (HDIs) can occur with the concomitant use of herb injectables and prescription drugs that are cleared by CYP450 enzymes, and further investigation is warrant for the clinical relevance of these interactions.

Effect of Wenjing Tongluo Decoction on VEGF/VEGFR2/ERK1/2 Signaling Pathway in Mice with Knee Osteoarthritis / 中国实验方剂学杂志

Objective:To investigate the possible mechanism of Wenjing Tongluo decoction （WTD） in alleviating articular cartilage defect in knee osteoarthritis （KOA） and delaying joint degeneration. Method:The KOA model was established by anterior cruciate ligament transection （ACLT）. Mice were classified into sham-operated group， model group， WTD high-dose and low-dose groups， and positive control group. Four weeks after modeling， WTD groups and the positive control group were given WTD （80， 20 g·kg^-1） and glucosamine sulfate capsules （0.29 g·kg^-1）， respectively， and the sham-operated group and model group received normal saline of the equivalent volume. After continuous intervention for 4 weeks， hemoxylin-eosin （HE） staining was used to observe the morphological changes of cartilage and Mankin scoring system was employed to score the knee cartilage. Western blot was combined with Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） to detect the protein and mRNA levels of vascular endothelial growth factor <italic>α</italic> （VEGFA）， vascular endothelial growth factor receptor 2 （VEGFR2）， extracellular signal-related kinase 1/2 （ERK1/2） and a disintegrin and metalloproteinase with thrombospondin motifs 4 （ADAMTS4）. Result:The Mankin score in the model group increased as compared with that in the sham-operated group （<italic>P</italic><0.01）. Compared with the model group， administration groups demonstrated alleviated articular cartilage defect and low Mankin score （<italic>P</italic><0.01）， but there was no statistical significance in Mankin score between the WTD groups and positive control group. The protein and mRNA levels of VEGFA， VEGFR2， ERK1/2， and ADAMTS4 in the model group were significantly higher than those in the sham-operated group （<italic>P</italic><0.01）. The protein expression of VEGFA and ERK1/2 was inhibited in each administration group as compared with that in the model group （<italic>P</italic><0.01）， and the inhibition in the positive control group was stronger than that in the WTD low-dose group （<italic>P</italic><0.05） but weaker than that in the WTD high-dose group （<italic>P</italic><0.01）. Glucosamine Sulfate capsules suppressed the expression of VEGFR2 and ADAMTS4 to the extent the same with low-dose WTD but weaker than the high-dose WTD （<italic>P</italic><0.05）. Conclusion:WTD can relieve the articular cartilage injury in KOA mice， and the mechanism may be related to VEGF/VEGFR2/ERK1/2 signaling pathway.

TAB1——Potential Biomarker of Paclitaxel Resistance in Ovarian Cancer Based on Label-Free Quantitative Proteomics / 中国实验方剂学杂志

Objective:To explore the potential targets and related mechanism involved in the paclitaxel resistance to ovarian cancer. Method:Ovarian cancer A2780 cells and A2780 paclitaxel-resistant cells （A2780/T） were treated by 2， 4， 8， 16， 32， 64， 128， 256 μmol·L^-1 paclitaxel （PTX） for 24 h or 48 h respectively <italic>in vitro</italic>. The proliferation rate of A2780 cells and A2780/T cells treated with paclitaxel was determined by methyl thiazolyl tetrazolium （MTT） colorimetric method assay. A2780 and A2780/T cells were analyzed by LC-MS/MS Label-Free quantitative proteomics to identify and screen differentially expressed proteins in the two groups of cells. Gene ontology （GO） annotation and Kyoto encyclopedia of genes and genomes （KEGG） pathway analysis were used to determine the potential biomarkers of paclitaxel resistance in ovarian cancer. Conventionally cultured A2780 cells were used as a control group， and A2780/T cells were treated with 0， 1， 4 μmol·L^-1 PTX. Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR） and Western blot methods were used to detect and verify the mRNA and protein expression levels of potential target transforming growth factor-<italic>β</italic>-activated kinase 1 binding protein 1 （TAB1） and its downstream related molecules transforming growth factor-<italic>β</italic>-activated kinase （TAK1） and p38. Result:After PTX treatment for 24 h and 48 h， the cell viability of A2780 and A2780/T cells decreased. The inhibitory rate of PTX on A2780 cells was significantly higher than that of A2780/T cells. In A2780 cells， the IC₅₀ of PTX treatment for 48 h was 0.002 μmol·L^-1， while in A2780/T cells， the IC₅₀of PTX was greater than the maximum concentration of 128 μmol·L^-1， indicating that A2780/T cells were resistant to PTX compared with A2780 cells. 441 differentially expressed proteins and 421 special differentially expressed proteins between A2780/T and A2780 cells were screened by label-free quantitative proteomic analysis. GO function enrichment analysis showed that the binding proteins accounted for the majority （80%） among the differentially expressed proteins. According to the results of KEGG pathway analysis and expression site analysis， TAB1 might be a potential biomarker in paclitaxel-resistant ovarian cancer. Compared with A2780 cells， mRNA and protein expression levels of TAB1 in A2780/T cells were significantly reduced （<italic>P</italic><0.01）. mRNA expression of TAK1 and p38 that interacted with TAB1 were also significantly reduced （<italic>P</italic><0.05， <italic>P</italic><0.01）， while there was no significant change in protein expression. Conclusion:TAB1 may be a potential biomarker of paclitaxel resistance to ovarian cancer ， and its mechanism may be related to the TAB1/TAK1/p38 MAPK pathway.

Genetic Polymorphism and Forensic Application of 47 Autosomal InDel Loci in 5 Chinese Ethnic Groups / 法医学杂志

Objective To investigate the population genetic data of 47 autosomal insertion/deletion （InDel） polymorphism genetic markers involved in AGCU InDel 50 kit in Guangdong Han, Guangxi Zhuang, Guangxi Yao, Guangxi Jing, and Guangxi Mulam, and to evaluate their application in forensic DNA identification. Methods Multiplex amplification of the 768 unrelated individuals from the 5 ethnic groups mentioned above was performed with the AGCU InDel 50 kit. Genotyping was carried out by 3500xL gene analyzer, population genetic parameters were gathered and polymorphism analysis was performed. Results No linkage disequilibrium was found among 47 autosomal InDel loci in the 5 ethnic groups. The distribution of genotype frequency of 47 autosomal InDel loci confirmed to the Hardy-Weinberg equilibrium in Guangdong Han and Guangxi Zhuang. Except for rs139934789, the other 46 loci confirmed to the Hardy-Weinberg equilibrium in Guangxi Yao, Guangxi Jing, and Guangxi Mulam. The results of genetic variation analysis among the populations showed that 1.12% of genetic variation was caused by ethnic group differences. The cumulative discrimination power of 47 autosomal InDel loci for the 5 ethnic groups were all above 0.999 999 999 999 999. The cumulative probability of exclusion for each ethnic group was less than 0.999 9. The two Y-InDels were identified in all male individuals and were absent in all female individuals. Conclusion Except for rs139934789, the other 46 InDel loci have a relatively good genetic polymorphism in the 5 Chinese ethnic groups, and can be used for forensic individual identification and as effective supplements for paternity testing.

Effect of Bushen Tongluo Formula on Osteoporosis of Zebrafish and Study on Autophagy Mechanism of Osteoclast / 中国实验方剂学杂志

Objective::To evaluate the intervention effect of Bushen Tongluo formula on zebrafish osteoporosis model and to clarify its regulation of autophagy mechanism on osteoclast differentiation. Method::The 260 young zebrafishes (3 dpf) were selected, the zebrafish osteoporosis model was established with 25 mmol·L-1 prednisolone (Pred) for 3 days, confirming the successful model by calcein staining. The zebrafishes were divided into control group, Pred group (25 mmol·L-1), etidronate disodium (ED) group (300 mg·L-1), Bushen (BS) group (180 mg·L-1), Tongluo (TL) group (30 mg·L-1), and Bushen Tongluo (BSTL) group (210 mg·L-1), 40 tails per group. After intervened with medicine for 4 days, the calcein staining was adopted to count the vertebral bone fluorescence area of zebrafish, Real-time PCR was adopted to detect the mRNA expression of akaline phosphatase (ALP), bone morphogenetic protein 2b (BMP-2b), runt-related protein 2 (Runx2) and cathepsin K (CTSK), phosphorus and tartrate-resistant acid phosphatase (TRAP), nuclear factor of activated T-cells, cytoplasmic-1 (NFATC-1). Divided RAW264.7 cells into control, Rankl induction (10 μg·L-1), BS (180 mg·L-1), TL (30 mg·L-1), and BSTL group (210 mg·L-1) after they were cultured to 80%-90% density. The expression of actin ring was detected by phalloidin cytoskeleton staining. The mRNA expression of TRAP, CTSK and autophagy-related genes 5 (ATG5), autophagy-related genes 7 (ATG7), and ubiquitin-binding protein p62 (p62) were detected by Real-time PCR. Result::Compared with the control group in vivo, the vertebral area and ALP, BMP-2b, and Runx2 expression of zebrafish in the Pred group were significantly decreased (P<0.01), and CTSK, TRAP, and NFATC-1 expression of zebrafish were significantly increased (P<0.01). Compared with the Pred group, the vertebral area of the TL (P<0.05) and BSTL group (P<0.01) increased significantly. The expressions of ALP, BMP-2b, and Runx2 in the BS, TL, and BSTL group were significant increased (P<0.01). The expression of CTSK, TRAP, and NFATC-1 in BSTL group were significant decreased (P<0.05, P<0.01). Compared with the control group in vitro, the number of actin rings (P<0.01) and CTSK, TRAP, ATG5 (P<0.01) expression and ATG7, p62 (P<0.01) expression were significantly increased in the Rankl-induced group. Compared with the Rankl induction group, the number of actin rings and CTSK and TRAP expression were significantly decreased in the BS, TL, and BSTL group (P<0.05, P<0.01), while the expression of ATG5, ATG7, and p62 were significant decreased in the BSTL group (P<0.05). Conclusion::BSTL can significantly improve prednisolone-induced zebrafish osteoporosis and inhibit osteoclast differentiation by reducing autophagy-related gene expression.

Analysis of Mechanism of Artesunate in Treatment of Liver Cancer Based on Metabolomics / 中国实验方剂学杂志

Objective: To study on the antitumor mechanism of artesunate in the treatment of liver cancer based on gas chromatography-mass spectrometry (GC-MS). Method: CellTiter-Glo® Luminescent Cell Viability Assay was used to detect activity of artesunate with different concentrations (0, 12.5, 25, 50, 100 μmol·L-1) on human liver cancer Huh7, SMMC-7721 cells for 24, 48, 72 h. GC-MS was employed to analyze the changes of metabolites of artesunate in two kinds of hepatoma cells (Huh7, SMMC-7721) for 24 h. The data was preprocessed by Postrun Analysis 4.41 workstation. Partial least squares-discriminant analysis (PLS-DA) was used to analyze two sets of differential metabolites and to analyze metabolic pathways of differential metabolites based on MetaboAnalyst 3.0 software. Result: Compared with the normal group, after two kinds of liver cancer cells was treated by artesunate, a total of 39 identical metabolites in the cells have undergone significant changes, which were mainly related to five metabolic pathways,including biosynthesis of aminoacyl-transfer RNA (tRNA), metabolism of alanine, aspartic acid and glutamic acid, metabolism of glycine, serine and threonine, metabolism of arginine and proline, metabolism of glutathione. Conclusion: Artesunate (12.5-100 μmol·L-1) can inhibit the growth of liver cancer cells (Huh7, SMMC-7721), it mainly involves five metabolic pathways, which may be the pathway of artesunate against liver cancer.

Application of zebrafish in field of pharmacy / 中国药理学通报

Zebrafish (Danio rerio) is a simple biological labora-tory animal, with similar biological structure and physiological functions to mammals. Almost transparent embryos, in vitro de-velopment of embryos and up to 87% of human genetic similari- ties enable a wide application of zebrafish in the field of pharma-ceutical research. In this paper, we have studied the application of zebrafish in drug metabolism, the research of Chinese medi-cine, the evaluation of drug toxicology and safety, the screening of drugs and the discovery of new drugs, the research of regener- ative drugs, so as to provide new ideas of zebrafish in the field of pharmacy.

Effects of ossotide injection on bone mineral density (BMD) , bone microstructure and biomechanical properties in ovariectomized rats / 中国比较医学杂志

Objective To observe the effect of ossotide injection on bone mineral density (BMD), bone microstructure and biomechanical properties and mRNA expression of small intestinal calcium binding protein (CaBp-D9K), and to study the mechanism of ossotide injection in the treatment of ovariectomized osteoporosis. Methods Forty-eight 3-month old SPF male rats with successful modeling (excision of bilateral ovaries) were randomly divided into the observation group and model group, 24 normal rats were divided into sham operation group (excised part of the mesenteric membrane), and 24 normal blank group. The blank group, sham operation group and model group were given normal saline, and the observation group was intragastrically given 1. 1 mg/kg ossotide. Two months after intervention, the bone volume (BV), trabecular bone volume (Tb. Th), trabecular number (Tb. N), trabecular separation (Tb. Sp), bone mineral density, bone biomechanics, serum 1,25(OH)2D3 levels and CaBp-D9K mRNA expression levels of small intestine were assessed and statistically analyzed. Results The bone mineral density, maximum load, fracture load, BV, Tb. Th, Tb. N, serum 1, 25 (OH ) 2D3 and intestinal CaBp-D9K mRNA expression level in the model group were significantly lower than those of the control group and sham operation group (P＜ 0. 05), Tb. Sp of the model group was significantly higher than that of the control group and sham operation group (P ＜ 0. 05 ). The bone mineral density, maximum load, fracture load, BV, Tb. Th, Tb. N, serum 1,25(OH)2D3 and intestinal CaBp-D9K mRNA expression level in the observation group were significantly higher than those of the model operation group, and the Tb. Sp of the observation group was lower than that of the model group (P ＜ 0. 05). Conclusions Ossotide injection treatment can reduce the degree of osteoporosis in ovariectomized rats, increasing intestinal CaBp-D9k mRNA expression and promoting intestinal calcium absorption may be its important mechanisms of action.

Effect of calcium dobesilate on vitreous hemorrhage in patients with PDR after pan retinal photocoagulation / 国际眼科杂志(Guoji Yanke Zazhi)

·AIM: To investigate the effect of calcium dobesilate on vitreous hemorrhage in patients with proliferative diabetic retinopathy ( PDR ) after pan retinal photocoagulation (PRP). ·METHODS:Totally 62 patients (30 cases with binocular lesions, 32 cases with monocular lesions, a total of 92 eyes) with PDR who were treated in our hospital from January 2015 to July 2017 were selected as the subjects. They were divided into the control group ( treated with pan retinal photocoagulation, n = 30, 17 cases with monocular lesions, 13 cases with binocular lesions, a total of 43 eyes ) and the study group ( treated with calcium dobesilate on the basis of treatment for the control group, n=32, 15 cases with monocular lesions, 17 cases with binocular lesions, a total of 49 eyes ). The recovery of visual acuity, blood rheology ( plasma viscosity, hematocrit, erythrocyte deformation index) and the incidence of complications such as vitreous hemorrhage in the two groups after surgery were observed. ·RESULTS: There was no significant difference between the two groups in the rate of excellent and good visual acuity, plasma viscosity, hematocrit or erythrocyte deformability index before treatment ( P>0. 05 ). After treatment, the rate of excellent and good visual acuity in the study group was significantly higher than that in the control group (P<0. 05). After treatment, the plasma viscosity and hematocrit decreased significantly while the erythrocyte deformability index significantly increased only in the study group, and changes of above -mentioned indexes in the study group were more obvious than those in the control group after treatment (P<0. 05). The incidence rate of vitreous hemorrhage and total incidence rate of complications in the study group were significantly lower than those in the control group ( P<0. 05). ·CONCLUSION: The application of calcium dobesilate in patients with PDR after pan retinal photocoagulation can effectively improve the recovery of visual acuity and reduce the incidence of complications such as vitreous hemorrhage. The mechanism may be related to effectively improving the hemodynamics.

Effects of interleukin-22 on the expression of CC chemokine ligand 27 in human epidermal keratinocytes / 中华皮肤科杂志

Objective To evaluate the effects of interleukin-22 (IL-22) on the expression of CC chemokine ligand 27 (CCL27) in human epidermal keratinocytes,and to explore its mechanism.Methods Immunohistochemical study was performed to determine the expression of CCL27 in skin lesions of 10 patients with psoriasis and skin tissues of 5 healthy controls.Cultured HaCaT cells were divided into 8 groups:control group treated with PBS,5 IL-22 groups treated with 12.5,25,50,100 and 200 μg/L IL-22 respectively,2 signaling pathway inhibition groups treated with 50 μrmol/L AG490 (JAK2/STAT3 signaling pathway inhibitor) or PD98059 (MAPK-ERK1/2 signaling pathway inhibitor) for 2 hours followed by the treatment with 50 μg/L IL-22 in the 5％ CO2 incubator at 37 ℃.After 24-hour cultivation,total proteins were extracted,and culture supernatants were collected,and both Western blot analysis and enzyme-linked immunosorbent assay (ELISA) were performed to determine the expression of CCL27.Results Immunohistochemical study showed that the expression of CCL27 was significantly higher in the skin lesions of the patients with psoriasis than in the skin tissues of the healthy controls.Western blot analysis revealed that the protein expression of CCL27 in the 12.5-,25-,50-,100-and 200-μg/L IL-22 groups was 0.491 ± 0.013,0.620 ± 0.019,0.623 ± 0.014,0.802 ± 0.052 and 1.138 ± 0.013 respectively,which were all higher than that in the control group (0.413 ± 0.013,all P ＜ 0.01).The expression of CCL27 was significantly lower in the IL-22 + AG490 group (0.411 ± 0.019) and IL-22 + PD98059 group (0.280 ± 0.012) than in the 50-μg/L IL-22 group (both P ＜ 0.01).ELISA also showed the same trend of changes in the level of CCL27 in the above groups as Western blot showed.Conclusion IL-22 can promote the expression of CCL27 in HaCaT cells,which may be associated with the MAPK-ERK 1/2 and JAK2/STAT3 signaling pathways.

A Reliability and Validity Study of the Korean versions of the Eating Disorder Examination Questionnaire version 6.0 (EDE-Q version 6.0) and the Clinical Impairment Assessment Questionnaire (CIA) / 정신신체의학

OBJECTIVES: The Eating Disorder Examination Questionnaire, version 6.0 (EDE-Q version 6.0) and the Clinical Impairment Assessment Questionnaire (CIA) measure attitudes and behavioral features of eating disorders and impairments secondary to eating disorders, respectively. The aims of this study were to examine the reliability and the validity of the Korean versions of the EDE-Q version 6.0 and the CIA. METHODS: Four hundred nineteen participants (370 female university students and 49 women with eating disorders) completed the EDE-Q version 6.0, the CIA, the Body Shape Questionnaire (BSQ) and the Weight Concern Scale (WCS). RESULTS: Excellent internal consistencies were obtained for the EDE-Q version 6.0 (Cronbach's α=0.92) and the CIA (Cronbach's α=0.91). Exploratory factor analysis of CIA extracted the 3 factors of personal, social, and cognitive impairments, as the original CIA had. The EDE-Q version 6.0 and the CIA were well correlated with the BSQ and the WCS, in respect to their contextually concordant variables. Patients with eating disorders had higher scores both in the EDE-Q 6.0 and the CIA than university women had, supporting good discriminant validity. CONCLUSIONS: The EDE-Q version 6.0 and the Korean versions of the CIA had adequate reliability and validity. These data will help clinicians and researchers to use the EDE-Q and the CIA in diagnosis, prevention and intervention of eating disorders in Korea.

Coupling of osteogenesis and angiogenesis in bones / 中国组织工程研究

BACKGROUND:In addition to a conduit system for oxygen,nutrients and waste products,blood vessels in bones manipulate multiple aspects of bone formation and provide niches for bone marrow hematopoietic stem cells.Coupling of osteogenesis and angiogenesis plays a crucial role in bone development,maturation,ageing and diseases.OBJECTIVE:To review the architecture of bone vasculature and process of angiogenesis,and then discuss the factors regulating coupling of osteogenesis and angiogenesis.METHODS:CBM and PubMed databases were retrieved using the keywords of "osteogenesis,blood vasculature,angiogenesis" in Chinese and English,respectively.High-quality articles related to the blood vasculature and angiogenesis in bones were included,and the repetitive studies were excluded.Finally 65 eligible articles were included to explore the coupling of osteogenesis and angiogenesis.RESULTS AND CONCLUSION:The formation and stabilization of the skeletal system rely on the coordination of several different types of cellular activities,especially endothelial cells as an important source of signals that regulate chondrocytes and osteogenesis-related cells,thus allowing the coupling of angiogenesis and osteogenesis during development and regeneration.Like other organs,the vascular system in the bones exhibits a typical hierarchy;the arterial trunk branches flow into the capillary network and then converge into the large veins in the center of the bone.Vascular endothelial growth factor,fibroblast growth factor 2,platelet-derived growth factor,matrix metalloproteinases,hypoxia,and Notch signaling pathway are involved in the angiogenesis in bones.The signaling pathways and related cytokines regulating the coupling of osteogenesis and angiogenesis will provide a new treatment strategy to battle bone diseases such as osteoporosis,bone tumor,bone fracture and bone repair.

Tolerance of rat bone marrow mesenchymal stem cells overexpressing human heme oxygenase 1 to ischemia/hypoxia injury / 中国组织工程研究

BACKGROUND:Under ischemia/hypoxia microenvironment,very low survival rate of transplanted bone marrow mesenchymal stem cells (BMSCs) in the host limits its efficacy in the treatment of acute myocardial infarction.OBJECTIVE:To explore the tolerance of human heme oxygenase 1 (hHO-1) gene modified rat BMSCs to ischemia/hypoxia injury.METHODS:The rat BMSCs were transfected with hHO-1 recombinant adenovirus.Western blot assay was used to determinate the optimal time of hHO-1 protein expression.hHO-1 modified rat BMSCs were cultured in hypoxia and serum-free conditions that simulated ischemia/hypoxia microenvironment in vivo.Cell counting kit-8 and trypan blue staining were performed to detect the survival rate of BMSCs at 12,24,48,72 hours after culture under the ischemia/hypoxia microenvironment.Flow cytometry was used to detect apoptosis in BMSCs at 24 hours after culture under the ischemia/hypoxia microenvironment.RESULTS AND CONCLUSION:The expression of hHO-1 protein was highest at 4 days after transfection.Under the ischemia/hypoxia microenvironment for 12,24,48,72 hours,the survival rates of transfected BMSCs were significantly higher as compared with the untransfected cells (P ＜ 0.05),shown by the cell counting kit-8 and trypan blue staining.In addition,the results from flow cytometry showed that there was a higher survival rate of transfected BMSCs than untransfected cells at 24 hours of culture under the ischemia/hypoxia microenvironment (P ＜ 0.05).To conclude,hHO-1 modified rat BMSCs have stronger tolerance to the ischemia/hvpoxia microenvironment.

Treatment of Ankylosing Spondylitis with Modified Jingang Pills / 中国中医药信息杂志

Ankylosing spondylitis has become a common disease, whether early stage of the low back pain or disability of the late stage of the disease has a serious impact on life quality. There is an urgent need of safe and effective treatment to treat the disease. This article introduced two cases of ankylosing spondylitis treated by modified Jingang Pills, with significant efficacy.

Cloning,expression and identification of the different fragments of polymorphic membrane protein I and its immunogenicity analysis of Chlamydia trachomatis serovar D / 中华传染病杂志

Objective To obtain the full length (FL ) and C‐terminal fragment of polymorphic membrane protein I (PmpI) of Chlamydia trachomatis serovar D ,and to study the immunogenicity of these proteins .Methods The target genes of PmpI‐FL and PmpI‐C were amplified by polymerase chain reaction (PCR) and inserted into the prokaryotic plasmid vector pGEX‐6P‐1 .The recombinant plasmids pGEX‐6P‐1/PmpI‐FL and pGEX‐6P‐1/PmpI‐C were separately transformed into Escherichia .coli ( E . coli) DH5αand were identified by enzyme digestion ,sequencing and PCR .After the identification ,the recombinant plasmids were separately transformed into E .coli BL21 and induced to express the proteins . The expected proteins were identified by Coomassie brilliant blue staining and Western blot ,then purified by glutathione S‐transferase (GST) MagBeads .The purified proteins were then injected into BALB/c mice to prepare the polyclonal antibodies against PmpI‐FL or PmpI‐C .Enzyme‐linked immune sorbent assay (ELISA) was used for the quantitative detection of the specific antibody .Results The lengths of cloned target genes PmpI‐FL and PmpI‐C were 2 659 bp and 1 195 bp ,respectively ,and the sequences were consistent with those of Chlamydia trachomatis serovar D in GenBank .The molecular masses of target proteins were 122 000 and 69 000 ,respectively ,which were confirmed by Coomassie brilliant blue staining and Western blot and then purified .The titers of the antibodies (anti‐PmpI‐FL and anti‐PmpI‐C) in sera of immunized mice detected by ELISA were 1∶12 800 and 1∶6 400 ,respectively .Conclusion The PmpI‐FL‐GST and PmpI‐C‐GST fusion proteins with high immunogenicity are successfully expressed and purified , which lays the foundation for further study .

The application of optical coherence tomography to measure the thickness of rabbit knee joint cartilage / 实用放射学杂志

Objective To study the feasibility of the measurement of the thickness of rabbit knee joint cartilage by comparing the optical coherence tomography (OCT)and the undecalcified frozen section method on respectively measure the thickness of New Zeal-and white rabbit’s knee cartilage.Methods 50 standardized cultivation,adult and male New Zealand white rabbits (100 knees) were selected in this study.The measurement point was at the knee weight-bearing area of the medial femoral condyle with a 2 milli-meter diameter trephine,and the cartilage thickness data at the same center point and ±0.5 mm from the center were obtained.OCT and freeze-sectioning method were adopted at each site,respectively.The difference between OCT and histological method was com-pared and Bland-Altman plot was constructed.Results The thickness results of the center,+0.5 mm,-0.5 mm points were (296.5± 1.6)μm,(302.6± 3.5)μm,(287.9±5.6)μm by OCT,(278.4±1.9)μm,(290.3±5.9)μm,(280.3±4.6)μm by freeze-sectio-ning method,respectively.In Bland-Altman diagram,mean difference and 95% confidence interval were 18.1 1 (1 6.65,1 9.56)μm, 12.4 (5.5,1 9.2)μm,7.4 (2.8,12.0)μm.Intra-class correlation coefficients(ICC)for resemblance between the 2 techniques were 0.93(95%CI:0.89-0.95,P <0.000 1),0.84(95%CI:0.77-0.89,P <0.000 1),0.91(95%CI:0.87 -0.94,P <0.000 1),respec-tively.Conclusion As compared with the measurement of undecalcified frozen section,OCT for the rabbits’knee joint cartilage thickness measurement is feasible with the advantages of noninvasiveness,good repeatability.OCT can provide data reference in ani-mal experiments of cartilage tissue engineering for articular cartilage defect repair.