Effects of human umbilical cord mesenchymal stem cell-derived exosomes in the rat osteoarthritis models.

Mesenchymal stem cells (MSCs) offer great potential for treatment of osteoarthritis (OA) by promoting articular cartilage regeneration via paracrine secretion of exosomes; however, the underlying mechanisms are not fully understood. This study aimed to explore the therapeutic effects of exosomes secreted by human umbilical cord-derived MSCs (hUC-MSCs) in rat models of OA and reveal the underlying mechanisms. UC-MSCs and UC-MSC-exosomes were prepared and identified by transmission electron microscopy and flow cytometry. IL-1ß-induced OA chondrocytes and the operation and collagenase-induced OA rat models were established. The results of micro-computed tomography, histology, and immunohistochemistry showed that UC-MSC-exosomes promoted cartilage regeneration in OA rats. ELISA results showed that the levels of synovial fluid cytokines, TNF-α, IL-1ß, and IL-6, were lower in exosome therapy group than control group in both OA rat models. Exosome treatment significantly downregulated the expression of MMP-13 and ADAMTS-5 in chondrocytes stimulated by IL-1ß, and upregulated collagen II expression. These findings suggest that hUC-MSC-exosomes offer a promising option for the therapy for OA.

Normal sperm head morphometric reference values in fertile Asian males.

ABSTRACT: Sperm head morphology is crucial for male factor infertility diagnosis and assessment of male reproductive potential. Several criteria are available to analyze sperm head morphology, but they are limited by poor methodology comparability and population applicability. This study aimed to explore comprehensive and new normal morphometric reference values for spermatozoa heads in fertile Asian males. An automated sperm morphology analysis system captured 23 152 stained spermatozoa from confirmed fertile males. Of these samples, 1856 sperm head images were annotated by three experienced laboratory technicians as "normal". We employed 14 novel morphometric features to describe sperm head size (head length, head width, length/width ratio, and girth), shape (ellipse intersection over union, girth intersection over union, short-axis symmetry, and long-axis symmetry), area (head, acrosome, postacrosomal areas, and acrosome area ratio), and degrees of acrosome and nuclear uniformity. This straight-forward method for the morphometric analysis of sperm by accurate visual measurements is clinically applicable. The measured parameters present valuable information to establish morphometric reference intervals for normal sperm heads in fertile Asian males. The presented detailed measurement data will be valuable for interlaboratory comparisons and technician training. In vitro fertilization and andrology laboratory technicians can use these parameters to perform objective morphology evaluation when assessing male fertilization potential.

External quality assessment scheme for sperm DNA fragmentation: a pilot study in China.

BACKGROUND: The aim of this article is to establish an external quality assessment (EQA) scheme for sperm Deoxyribonucleic acid (DNA) fragmentation (SDF) detection, and to assess the feasibility of the scheme. In addition, this article provides some case analysis of abnormal results in order to really help improve the performance of the laboratory. RESULTS: In 2021 and 2022, 10 and 28 laboratories in China volunteered to participate in the EQA program respectively. Two samples were selected for EQA each year, a large spread of results was obtained for the four samples, and the highest values were 13.7, 4.2, 8.0 and 4.0 times the lowest respectively. The coefficients of variation (CVs) were very high for the four samples, at 46.6%, 30.1%, 26.7% and 30.3%, respectively. The CVs of the samples with high SDF values were lower than those of the samples with low SDF values. There was no significant difference between the results of sperm chromatin structure assay (SCSA) and sperm chromatin dispersion (SCD). For the 10 laboratories that participated in EQA in 2021 and 2022, the CVs of low SDF value samples and high SDF value samples decreased from 46.6% and 30.1% in 2021 to 32.5% and 22.7% in 2022, respectively. CONCLUSION: This is the first study to evaluate the EQA program on SDF, which involved a number of laboratories and was demonstrated to be feasible. It is recommended that all laboratories participate in the EQA of SDF to ensure the accuracy of the results.

RéSUMé: CONTEXTES: L'objectif de cet article est d'établir un système externe d'évaluation de la qualité (EEQ) pour la détection de la fragmentation de l'ADN des spermatozoïdes (SDF) et d'évaluer la faisabilité de ce système. En outre, cet article fournit une analyse de cas de résultats anormaux afin d'aider réellement à améliorer les performances du laboratoire. RéSULTATS: En 2021 et 2022, respectivement 10 et 28 laboratoires en Chine se sont portés volontaires pour participer au programme EEQ. Deux échantillons ont été sélectionnés chaque année pour l'EEQ ; un large éventail de résultats a été obtenu pour les quatre échantillons, et les valeurs les plus élevées étaient respectivement de 13,7, 4,2, 8,0 et 4,0 fois les plus faibles. Les coefficients de variation (CV) étaient très élevés pour les quatre échantillons, soit respectivement 46,6 %, 30,1 %, 26,7 % et 30,3 %. Les CV des échantillons avec des valeurs de SDF élevées étaient inférieurs à ceux des échantillons avec de faibles valeurs de SDF. Il n'y avait pas de différence significative entre les résultats du test de structure de la chromatine des spermatozoïdes (SCSA) et ceux de la dispersion de la chromatine des spermatozoïdes (SCD). Pour les 10 laboratoires qui ont participé à l'EEQ en 2021 et 2022, les CV des échantillons à faible valeur de SDF et ceux des échantillons à valeur élevée de SDF ont diminué, passant respectivement de 46,6 % et 30,1 % en 2021 à 32,5 % et 22,7 % en 2022. CONCLUSIONS: Il s'agit de la première étude à évaluer le programme externe d'évaluation de la qualité (EEQ) de l'analyse de la SDF, qui a impliqué un certain nombre de laboratoires, et qui s'est avéré réalisable. Il est recommandé que tous les laboratoires participent à l'EEQ de la SDF afin d'en assurer l'exactitude des résultats.

Periplaneta americana extract CⅡ-3 induces senescence of leukemia K562 cells via SIRT1/mTOR signaling pathway / 中国中药杂志

This study aims to investigate the role of slient mating-type information regulation 2 homolog 1(SIRT1)/tuberous sclerosis complex 2(TSC2)/mammalian target of rapamycin(mTOR) signaling pathways in the Periplaneta americana extract CⅡ-3-induced senescence of human leukemia K562 cells. K562 cells were cultured in vitro and treated with 0(control), 5, 10, 20, 40, 80, and 160 μg·mL~(-1) of P. americana extract CⅡ-3. Cell counting kit-8(CCK-8) and flow cytometry were employed to examine the proliferation and cell cycle of the K562 cells. Senescence-associated β-galactosidase stain kit(SA-β-gal) was used to detect the positive rate of senescent cells. Mitochondrial membrane potential was detected by flow cytometry. The relative mRNA level of telomerase reverse transcriptase(TERT) was determined by fluorescence quantitative PCR. The mRNA and protein levels of SIRT1, TSC2, and mTOR were determined by fluorescence quantitative PCR and Western blot, respectively. The results showed that CⅡ-3 significantly inhibited the proliferation of K562 cells and the treatment with 80 μg·mL~(-1) CⅡ-3 for 72 h had the highest inhibition rate. Therefore, 80 μg·mL~(-1) CⅡ-3 treatment for 72 h was selected as the standard for subsequent experiments. Compared with the control group, CⅡ-3 increased the proportion of cells arrested in G_0/G_1 phase, decreased the proportion of cells in S phase, increased the positive rate of SA-β-Gal staining, elevated the mitochondrial membrane potential and down-regulated the mRNA expression of TERT. Furthermore, the mRNA expression of SIRT1 and TSC2 was down-regulated, while the mRNA expression of mTOR was up-regulated. The protein expression of SIRT1 and p-TSC2 was down-regulated, while the protein expression of p-mTOR was up-regulated. The results indicated that P. americana extract CⅡ-3 induced the senescence of K562 cells via the SIRT1/mTOR signaling pathway.

Effect of prophylactic anticoagulation duration on venous thrombosis after total hip/knee arthroplasty / 中国临床药理学与治疗学

AIM: By assessing the impact of prolonged prophylactic anticoagulation on venous thromboembolism in patients undergoing total hip/ knee arthroplasty, we dared to hope to further clarify whetherprolonged prophylactic anticoagulation duration can benefit patients undergoing total hip/ knee arthroplasty. METHODS: The incidence of venous thromboembolism and bleeding events within 90 days of total hip/knee arthroplasty in patients who underwent total hip/knee arthroplasty in the department of orthopaedic surgery was retrospectively analyzed from January 2019 to April 2022. The Kaplan-Meier method survival curve was used to determine whether there is a relationship between the duration of prophylactic anticoagulation and the incidence of postoperative bleeding. RESULTS: A total of 115 patients undergoing primary total hip/knee surgery from January 2019 to April 2022, were enrolled in this study. Among them, there were 38 cases in the short-term prophylactic anticoagulation group and 77 cases in the extended prophylactic anticoagulation group. There were 23 cases (20%) of venous thromboembolism within 90 days after surgery, of which 12 cases (31.58%) were in the short-term anticoagulation group and 11 cases (14.29%) were in the extended anticoagulation group, and there was a statistical difference in the incidence of venous thromboembolism within 90 days after surgery between the two groups in terms of the duration of anticoagulation prevention. CONCLUSION: The results show a significant correlation between the duration of prophylactic anticoagulation and the incidence of venous thromboembolism within 90 days after total hip/knee arthroplasty, which suggests that prophylactic anticoagulation for 15-35 days after undergoing total hip arthroplasty or total knee arthroplasty reduces the incidence of postoperative VTE, and there is no significant difference in bleeding risk depending on the duration of anticoagulant prophylaxis.

Semen parameters are seriously affected in acephalic spermatozoa syndrome.

BACKGROUND: Previous studies have reported that some patients with headless spermatozoa have poor semen quality, but there has been no published systematic analysis of semen quality in patients with different proportions of headless spermatozoa in semen. We aimed to explore the association of acephalic spermatozoa syndrome and semen quality in men with distinct proportions of headless spermatozoa. MATERIAL AND METHODS: Semen parameter values in patients for whom headless spermatozoa were found in the ejaculates was studied and compared to that of 413 age-matched prenatal examination patients. All semen samples were analyzed following the same methodology in a single laboratory. RESULTS: All semen parameter values except semen volume were negatively (P < 0.05) correlated with the proportion of headless spermatozoa. The semen samples were divided into four groups on the basis of the proportion of headless spermatozoa (PHS) as follows: 0 < PHS ≤ 5% (n = 172, Group A1); 5 < PHS ≤ 10% (n = 76, Group A2); 10 < PHS ≤ 20% (n = 71, Group B); and PHS > 20% (n = 71, Group C). In Group A1, only one semen parameter value (progressive motility) was lower than those of the control group, but in Group A2, this increased to five (sperm vitality, normal sperm morphology, sperm motility, VCL (curvilinear velocity) and ALH (amplitude of lateral head displacement)). Worse still, all semen parameter values were significantly lower in Group B and Group C than in the control group (P < 0.05). CONCLUSIONS: Semen samples containing headless spermatozoa tend to have lower quality than samples without headless spermatozoa. Increases in the proportion of headless spermatozoa in semen are associated with decreased semen quality. We suggest that headless spermatozoa should be seriously assessed and accurately counted in semen analysis, especially for ejaculate in which the proportion of headless spermatozoa exceeds 5%.

RéSUMé: CONTEXTE: Des études antérieures ont rapporté que certains patients qui avaient des spermatozoïdes sans tête présentaient une mauvaise qualité du sperme, mais il n'y a eu aucune analyse systématique publiée sur la qualité du sperme chez les patients ayant des proportions différentes de spermatozoïdes sans tête dans leur sperme. Nous avons cherché à explorer l'association entre syndrome des spermatozoïdes acéphaliques et qualité du sperme chez les hommes ayant des proportions distinctes de spermatozoïdes sans tête. Les valeurs des paramètres du sperme chez les patients pour lesquels des spermatozoïdes sans tête ont été trouvés dans l'éjaculat ont été étudiées et comparées à celles de 413 patients consultant pour un examen prénatal, appariés sur l'âge. Tous les échantillons de sperme ont été analysés selon la même méthodologie dans un seul laboratoire. RéSULTATS: Toutes les valeurs des paramètres du sperme, à l'exception du volume de sperme, étaient négativement (P < 0,05) corrélées avec la proportion de spermatozoïdes sans tête. Les échantillons de sperme ont été divisés en quatre groupes sur la base de la proportion de spermatozoïdes sans tête (PHS) comme suit: 0 < PHS ≤ 5% (n = 172, groupe A1); 5 < PHS ≤ 10% (n = 76, groupe A2); 10 < PHS ≤ 20% (n = 71, groupe B); et PHS > 20% (n = 71, groupe C). Dans le groupe A1, une seule valeur de paramètre de sperme (motilité progressive) est. inférieure à celle du groupe témoin, mais dans le groupe A2, le nombre s'élève à cinq (vitalité des spermatozoïdes, morphologie normale des spermatozoïdes, mobilité des spermatozoïdes, VCL (vitesse linéaire curviligne) et ALH (amplitude du déplacement latéral de la tête)). Pire encore, toutes les valeurs des paramètres du sperme étaient significativement plus faibles dans les groupes B et C que dans le groupe témoin (P < 0,05). CONCLUSIONS: Les échantillons de sperme contenant des spermatozoïdes sans tête ont tendance à avoir une qualité inférieure à celle des échantillons dépourvus de spermatozoïdes sans tête. L'augmentation de la proportion de spermatozoïdes sans tête dans le sperme est. associée à une réduction de la qualité du sperme. Nous suggérons que les spermatozoïdes sans tête devraient être sérieusement évalués et comptés avec précision dans l'analyse du sperme, en particulier pour l'éjaculat dans lequel la proportion de spermatozoïdes sans tête dépasse 5%.

Nurses' Workplace Social Capital and the Influence of Transformational Leadership: A Theoretical Perspective.

Workplace social capital is the relational network, created by respectful interactions among members of a workforce, can contribute to the formation of a wholesome psychological work environment in an organization. Nurses' workplace social capital is a derivative of the workplace social capital, formed because of the complex interactions among the nursing and between the other healthcare professionals. Transformational leadership is a style of leadership that addresses the emotional wellbeing of its workforce and inspires shared group ethics, norms, and goals. The philosophy of transformational leadership is grounded on the premise of workforce as human beings with specific needs. Transformational leadership has been confirmed as a strong predictor of nurses' workplace social capital. Meanwhile, it is of an academic and/or healthcare industry operational value to scholarly assess and discern the theoretical influence of transformational leadership on nurses' workplace social capital. In this paper, we have attempted to explore the associations between transformational leadership and nurses' workplace social capital from a theoretical perspective. We have discussed the importance of each sub-dimension of transformational leadership (modeling the way, inspiring a shared vision, challenging the process, enabling others to act and encouraging the heart) in building up the social capital relational network. Finally, we have proposed a graphic framework of our analysis to facilitate understanding of the associations between the transformational leadership and nurses' workplace social capital, in formation of a healthy work environment which is the foundation for efficiency and productivity of the workforce.

Retrospective analysis of bladder cancer morphology and depth of invasion under cystoscopy.

BACKGROUND: The pathological diagnosis of bladder cancer workup relies on cystoscopy, however, due to sampling restriction, the depth of local invasion is often understaged. METHODS: A total of 386 patients with bladder urothelial carcinoma underwent follow-up. The data collected included age, sex, tumor size, surgical options, histologic grade, invasive depth, lymph node metastasis, and oncological outcomes, and the patients were divided into coral-like and crumb-like groups. These data were analyzed with the chi-square test, binary logistic regression, Kaplan-Meier analysis, univariable and multivariable logistic regression and Spearman correlation test. RESULTS: Bladder tumor morphology was moderately correlated with invasion depth (ρ = 0.492, p < 0.001; Spearman correlation), which was associated with invasion status (HR = 8.27; 95% CI 4.3-15.79, p < 0.001). Tumor morphology was not an independent factor for OS but was associated with PFS. Outer invasion depth was an independent factor that was significantly associated with inferior OS and PFS. CONCLUSIONS: Tumor morphology (coral-like and crumb-like) under cystoscopy was related to the depth of invasion. The outer invasive depth of BC was an independent factor that was significantly associated with inferior OS and PFS.

Optimization of UC-MSCs cold-chain storage by minimizing temperature fluctuations using an automatic cryopreservation system.

The effective long-term cryopreservation of human mesenchymal stem cells is an essential prerequisite step and represents a critical approach for their sustained supply in basic research, regenerative medicine, and tissue engineering applications. Off-the-shelf availability of human umbilical cord-derived mesenchymal stromal cells (UC-MSCs) for regenerative medicine application requires the development of nontoxic, safe, and efficient protocols for cryopreservation. In the long-term low-temperature storage process of cells, traditional manual storage has a great impact on cell activity, recovery, and function due to repeated exposure of cells to room temperature. To minimize the effect of fluctuation in ambient temperature on stored cells, we designed an automatic cryopreservation system that handles cells under controlled temperatures. In this work, UC-MSCs were utilized to investigate and compare the influence of manual and automatic cryopreservation approaches. To simulate the manual process, the UC-MSCs were transferred back and forth repeatedly (up to 400 times) between the liquid nitrogen (LN2) tank (-150 °C) and room temperature by a robotic arm. Similarly, the UC-MSCs from the same batch were collected and transferred repeatedly between two storage units by the automatic cryopreservation system, where the cells were maintained below-150 °C throughout the cold chain process. Viability, percent recovery, adherence capability, cell proliferation, and multilineage differentiation ability were assessed for UC-MSCs. Compared to the manual approach, UC-MSCs handled by the automatic system demonstrated higher viability, percent recovery, and cell proliferation, as well as improved adherence to culture plate with greater potential in multilineage differentiation after 400 temperature cycles. The described entire cold chain system may provide a powerful tool to develop safe, reliable and efficient protocols for manufacturing and banking of UC-MSCs, improving their off-the-shelf availability for regenerative medicine applications.

Emergence of blaNDM-5 in Enterobacteriaceae Isolates from Companion Animals in Guangzhou, China.

The occurrence and characterization of carbapenemase-producing Enterobacteriaceae from companion animals in Guangzhou, China, are investigated. Six isolates (2.3%, 6/257) were positive for blaNDM-5, that is, one Enterobacter cloacae, one Citrobacter freundii, and four Escherichia coli. Three E. coli isolates obtained from the same animal hospital were ST410 and showed identical pulse field gel electrophoresis pattern, resistance profiles, and resistance genes. blaNDM-5 was located on IncX3 (n = 5) and IncK2 (n = 1) plasmid, respectively. The presence of carbapenemase-producing Enterobacteriaceae among companion animals needs continued surveillance.

Acupoint Catgut Embedding Alleviates Insomnia in Different Chinese Medicine Syndrome Types: A Randomized Controlled Trial / 中国结合医学杂志

OBJECTIVE@#To investigate the effects and safety of catgut embedding on alleviating insomnia.@*METHODS@#Totally 510 patients with insomnia were divided into 5 Chinese medicine (CM) syndrome types: Xin (Heart) and Pi (Spleen) deficiency, yin deficiency with excess fire, Xin and gut qi deficiency, Wei (Stomach) disorder, and qi and blood deficiency, respectively. These 5 types of patients were randomly assigned to a catgut embedding group, an acupuncture group or a medication group (30 cases in Xin and Pi deficiency type, Wei disorder type, Xin and gut qi deficiency type, respectively; 40 cases in yin deficiency with excess fire type and qi and blood deficiency type, respectively). In the catgut embedding group, patients were treated by implanting catgut into acupoints once every 10 days for a total of 30 days. In the acupuncture group, patients were treated with acupuncture once per day over 30 days (excluding weekends); and patients in the medication group took 1 mg Eurodin Tablet orally every night for 30 days. Pittsburgh Sleep Quality Index (PSQI) was evaluated before treatment, on 30 and 60 days after the first treatment, respectively. The International Unified Sleep Efficiency Value (IUSEV) was measured at 30 and 60 days. The safety was evaluated after treatment and adverse events were analyzed.@*RESULTS@#The objective PSQI scores including subjective sleep quality, sleep latency, sleep duration, habitual sleep efficiency, sleep disturbance, daytime dysfunction, and total scores at 30 days were significantly improved compared with pre-treatment in the catgut embedding and acupuncture groups (P<0.01 or P<0.05). At 30 days, the PSQI scores in catgut embedding group were superior to the medication group in the patients with each type of insomnia, with the exception of sleep duration (P<0.01 or P<0.05). At 60 days, significant differences were found between the catgut embedding group and the medication group (P<0.01 for all indices). The IUSEV scores in the catgut embedding group were significantly higher than the acupuncture group at 60 days, and the scores in acupuncture group were higher than the medication group at 30 days (P<0.05 for all types). No severe adverse events were found in this study.@*CONCLUSIONS@#Acupoint catgut embedding and acupuncture were more effective than medication in alleviating insomnia syndrome in different Chinese medicine syndrome type. However, the sustained effects of acupoint catgut embedding were superior to acupuncture.

Clonal Spread of Escherichia coli ST93 Carrying mcr-1-Harboring IncN1-IncHI2/ST3 Plasmid Among Companion Animals, China.

The purpose of this study was to investigate the occurrence of plasmid-mediated colistin resistance gene mcr-1 in Enterobacteriaceae isolates from companion animals in Guangzhou, China. Enterobacteriaceae isolated from 180 samples collected from cats and dogs were screened for mcr-1 by PCR and sequencing. MCR-1-producing isolates were further characterized by multilocus sequence typing and pulsed-field gel electrophoresis (PFGE). Plasmid characterization was performed by conjugation, replicon typing, S1-PFGE, and Southern blot hybridization. Plasmid pHN6DS2 as a representative IncN1-IncHI2/ST3 plasmid from ST93 E. coli was fully sequenced. pHN6DS2-like plasmids were screened by PCR-mapping and sequencing. The mcr-1 gene was detected in 6.25% (8/128) Escherichia coli isolates, of which, five belonged to E. coli ST93 and had identical PFGE patterns, resistance profiles and resistance genes. mcr-1 genes were located on ∼244.4 kb plasmids (n = 6), ∼70 kb plasmids, and ∼60 kb plasmids, respectively. Among them, five mcr-1-carrying plasmids were successfully transferred to recipient by conjugation experiments, and were classified as IncN1-IncHI2/ST3 (∼244.4 kb, n = 4, all obtained from E. coli ST93), and IncI2 (∼70 kb, n = 1), respectively. Plasmid pHN6DS2 contained a typical IncHI2-type backbone, with IncN1 segment (ΔrepA-Iterons I-gshB-ΔIS1294) inserted into the multiresistance region, and was similar to other mcr-1-carrying IncHI2/ST3 plasmids from Enterobacteriaceae isolates of various origins in China. The remaining five mcr-1-bearing plasmids with sizes of ∼244.4 kb were identified to be pHN6DS2-like plasmids. In conclusion, clonal spread of ST93 E. coli isolates was occurred in companion animals in Guangzhou, China.

Influence of Feeding Type and Nosema ceranae Infection on the Gut Microbiota of Apis cerana Workers.

The gut microbiota plays an essential role in the health of bees. To elucidate the effect of feed and Nosema ceranae infection on the gut microbiota of honey bee (Apis cerana), we used 16S rRNA sequencing to survey the gut microbiota of honey bee workers fed with sugar water or beebread and inoculated with or without N. ceranae. The gut microbiota of A. cerana is dominated by Serratia, Snodgrassella, and Lactobacillus genera. The overall gut microbiota diversity was show to be significantly differential by feeding type. N. ceranae infection significantly affects the gut microbiota only in bees fed with sugar water. Higher abundances of Lactobacillus, Gluconacetobacter, and Snodgrassella and lower abundances of Serratia were found in bees fed with beebread than in those fed with sugar water. N. ceranae infection led to a higher abundance of Snodgrassella and a lower abundance of Serratia in sugar-fed bees. Imputed bacterial Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways showed the significant metagenomics functional differences by feeding and N. ceranae infections. Furthermore, A. cerana workers fed with sugar water showed lower N. ceranae spore loads but higher mortality than those fed with beebread. The cumulative mortality was strongly positive correlated (rho = 0.61) with the changes of overall microbiota dissimilarities by N. ceranae infection. Both feeding types and N. ceranae infection significantly affect the gut microbiota in A. cerana workers. Beebread not only provides better nutrition but also helps establish a more stable gut microbiota and therefore protects bees in response to N. ceranae infection. IMPORTANCE The gut microbiota plays an essential role in the health of bees. Scientific evidence suggests that diet and infection can affect the gut microbiota and modulate the health of the gut; however, the interplay between those two factors and the bee gut microbiota is not well known. In this study, we used a high-throughput sequencing method to monitor the changes of gut microbiota associated with both feeding types and Nosema ceranae infection. Our results showed that the gut microbiota composition and diversity of Asian honey bee were significantly associated with both feeding types and the N. ceranae infection. More interestingly, bees fed with beebread showed higher microbiota stability and lower mortality rates than those fed with sugar water when infected by N. ceranae. Those data suggest that beebread has the potential not only to provide better nutrition but also help to establish a more stable gut microbiota to protect bees against N. ceranae infection.

Evolution and Comparative Genomics of F33:A-:B- Plasmids Carrying blaCTX-M-55 or blaCTX-M-65 in Escherichia coli and Klebsiella pneumoniae Isolated from Animals, Food Products, and Humans in China.

To understand the underlying evolution process of F33:A-:B- plasmids among Enterobacteriaceae isolates of various origins in China, the complete sequences of 17 blaCTX-M-harboring F33:A-:B- plasmids obtained from Escherichia coli and Klebsiella pneumoniae isolates from different sources (animals, animal-derived food, and human clinics) in China were determined. F33:A-:B- plasmids shared similar plasmid backbones comprising replication, leading, and conjugative transfer regions and differed by the numbers of repeats in yddA and traD and by the presence of group II intron, except that pHNAH9 lacked a large segment of the leading and transfer regions. The variable regions of F33:A-B- plasmids were distinct and were inserted downstream of the addiction system pemI/pemK, identified as the integration hot spot among F33:A-B- plasmids. The variable region contained resistance genes and mobile elements or contained segments from other types of plasmids, such as IncI1, IncN1, and IncX1. Three plasmids encoding CTX-M-65 were very similar to our previously described pHN7A8 plasmid. Four CTX-M-55-producing plasmids contained multidrug resistance regions related to that of F2:A-B- plasmid pHK23a from Hong Kong. Five plasmids with IncN and/or IncX replication regions and IncI1-backbone fragments had variable regions related to those of pE80 and p42-2. The remaining five plasmids with IncN replicons and an IncI1 segment also possessed closely related variable regions. The diversity in variable regions was presumably associated with rearrangements, insertions, and/or deletions mediated by mobile elements, such as IS26 and IS1294IMPORTANCE Worldwide spread of antibiotic resistance genes among Enterobacteriaceae isolates is of great concern. F33:A-:B- plasmids are important vectors of resistance genes, such as blaCTX-M-55/-65, blaNDM-1, fosA3, and rmtB, among E. coli isolates from various sources in China. We determined and compared the complete sequences of 17 F33:A-:B- plasmids from various sources. These plasmids appear to have evolved from the same ancestor by mobile element-mediated rearrangement, acquisition, and/or loss of resistance modules and similar IncN1, IncI1, and/or IncX1 plasmid backbone segments. Our findings highlight the evolutionary potential of F33:A-:B- plasmids as efficient vectors to capture and diffuse clinically relevant resistance genes.

Study on Enrichment of Total Flavonoids from Licorice Residue by Chemical Conversion Method Based on Fingerprint and Quantitative Analysis of Multi-components with a Single-marker Technique / 中国中医药信息杂志

Objective To establish a combined quality evaluation model of fingerprint and quantitative analysis of multi-components with a single-marker (QAMS) to analyze the total flavonoids from licorice residue by the chemical conversion method; To provide technical support for quality control in production. Methods Total flavonoids of breaking cell wall and enriching were taken as the object of study to establish fingerprint. With liquiritin as internal reference, the relative correction factors of isoliquiritin, glycyrrhizin, isoliquiritigenin and glycyrrhizic acid were established respectively, and the contents were determined. Meanwhile, the calculated values were compared with the measured value by external standard method to verify the practicability and stability of QAMS. Results The HPLC fingerprint of total flavonoids from licorice residue was established. 11 common peaks were identified, and 5 common peaks were identified, and the similarity of the 10 extracts was >0.99; the relative error between the calculated results of QAMS and the measured values of the external standard method was <4%; the RSD of relative correction factor calculated by the multiple concentration method was <2%. Conclusion The method is accurate, reliable, specific, and stable, with good repeatability, which can be used for the quality control of total flavonoids from licorice residue.

Characterization of oqxAB in Escherichia coli Isolates from Animals, Retail Meat, and Human Patients in Guangzhou, China.

The purpose of this study was to investigate the prevalence and genetic elements of oqxAB among Escherichia coli isolates from animals, retail meat, and humans (patients with infection or colonization) in Guangzhou, China. A total of 1,354 E. coli isolates were screened for oqxAB by PCR. Fifty oqxAB-positive isolates were further characterized by pulsed-field gel electrophoresis (PFGE), multilocus sequence typing (MLST), S1-PFGE, genetic environment analysis, plasmid replicon typing, and plasmid sequencing. oqxAB was detected in 172 (33.79%), 60 (17.34%), and 90 (18.07%) E. coli isolates from animal, food, and human, respectively. High clonal diversity was observed among oqxAB-positive isolates. In 21 oqxAB-containing transformants, oqxAB was flanked by two IS26 elements in the same orientation, formed a composite transposon Tn6010 in 19 transformants, and was located on plasmids (33.3~500 kb) belonging to IncN1-F33:A-:B- (n = 3), IncHI2/ST3 (n = 3), F-:A18:B- (n = 2), F-:A-:B54 (n = 2), or others. Additionally, oqxAB was co-located with multiple resistance genes on the same plasmid, such as aac(6')-Ib-cr and/or qnrS, which were identified in two F-:A18:B- plasmids from pigs, and blaCTX-M-55, rmtB, fosA3, and floR, which were detected in two N1-F33:A-:B- plasmids from patients. The two IncHI2/ST3 oqxAB-bearing plasmids, pHNLDF400 and pHNYJC8, which were isolated from human patient and chicken meat, respectively, contained a typical IncHI2-type backbone, and were similar to each other with 2-bp difference, and also showed 99% identity to the Salmonella Typhimurium oqxAB-carrying plasmids pHXY0908 (chicken) and pHK0653 (human patient). Horizontal transfer mediated by mobile elements may be the primary mechanism underlying oqxAB spread in E. coli isolates obtained from various sources in Guangzhou, China. The transmission of identical oqxAB-carrying IncHI2 plasmids between food products and humans might pose a serious threat to public health.

[Influential factors analysis of positive psychology in patients with oral and maxillofacial trauma].

PURPOSE: To investigate the positive psychological reaction of patients with oral and maxillofacial trauma and related factors. METHODS: One hundred and five hospitalized patients with oral and maxillofacial trauma were investigated by self-designed general data questionnaire, positive psychological scale posttraumatic growth evaluation of quantitative PTG, and self-image questionnaire. SPSS 18.0 software package was used to analyze the data. RESULTS: Positive psychological score of the patients was 56.01±17.322, and self-image average score was 51.33±7.306. There were significant differences between male and female patients after trauma in new possibilities, personal power, self transformation and personal feeling (P<0.05); there was no significant difference between different ages in positive psychological reaction.With the improvement of educational level of patients, better personal power (P=0.031) and self transformation (P=0.01), and more positive psychological reaction were observed; Posttraumatic positive psychology of patients was negatively correlated with self-image score (r=-0.318, P<0.001). CONCLUSIONS: The male patients with oral and maxillofacial trauma have more positive attitude than female. With the improvement of educational level, more positive psychological reaction was documented in term of personal strength, self-transformation,but no obvious change in relationship with others, new possibilities and personal feeling. The better self image, the more positive psychological reaction was displayed.

Phenols from Plantaginis Semen / 中成药

AIM To study the phenols from Plantaginis Semen.METHODS The 65％ and 95％ ethanol extracts of Plantaginis Semen were isolated and purified by macroporous resin,silica,ODS,Sephadex and preparative column,then the structures of obtained compounds were identified by physicochemical properties and spectral data.RESULTS Nine compounds were isolated and identified as (+)-(7R,7'R,8S,8'S)-neo-olivil (1),erythro-guaiacylglycerol-β-ferulic acid ether (2),eriodictyol (3),luteolin (4),chrysoeriol (5),hydroxytyrosol (6),4-(3,4-dihydroxyphenyl)-(E)-3-buten-2-one (7),ferulic acid (8),5,7-dihydroxychromone (9).CONCLUSION Compounds 2-3,6-7 and 9 are isolated from genus Plantago for the first time,compound 5 is first obtained from this plant.

Simultaneous Determination of Liquiritin, Isoliquiritin, Glycyrrhizin, Isoliquiritigenin and Glycyrrhizic Acid in Licorice Extract by HPLC Dual Wavelength Spectrophotometry / 中国中医药信息杂志

Objective To establish a method for the simultaneous determination of liquiritin, isoliquiritin, glycyrrhizin, isoliquiritigenin and glycyrrhizic acid in licorice extract. Methods Liquiritin, isoliquiritin, glycyrrhizin, isoliquiritigenin and glycyrrhizic acid in licorice extract were determined by HPLC dual wavelength spectrophotometry. Symmetry C18 column (4.6 mm × 250 mm, 5 μm) was used. The mobile phase was acetonitrile (A) - 0.085％ phosphoric acid water (B), ingradient elution mode (0–8 min, 81％ B; 8–35 min, 81％→50％ B; 35–60 min, 50％ B) with the flow rate of 1.0 mL/min. The sample size was 10 μL, and column temperature was room temperature. Dual wavelength detection, λ1=237 nm, λ2=254 nm. Results Liquiritin, isoliquiritin, glycyrrhizin, isoliquiritigenin and glycyrrhizic acid were linear in the ranges of 0.0408–0.816 μg, 0.0528–1.056 μg, 0.0224–0.448 μg, 0.0212–0.424 μg, and 0.0448–0.896 μg, respectively. The average recovery was 98.69％, 98.31％, 99.10％, 98.55％, and 99.14％, respectively; RSD was 1.39％, 1.29％, 1.78％, 2.14％, and 1.15 ％, respectively. Conclusion The method is accurate, reliable and specific. The results are stable with good repeatability. It can be used for the determine of above 5 components in licorice extract.

Effect of evodiamine in inducing apoptosis of gastric cancer SGC-7901 cells through mTOR signal pathway / 中国中药杂志

Evodiamine is one of the most important antitumor alkaloid from evodiamine. This study focused on the mechanism of evodiamine in inducing apoptosis of gastric cancer SGC-7901 cells through mammalian target of rapamycin (mTOR) signal pathway, in order to explore its antitumor mechanism and lay a foundation for clinical treatment of gastric cancer. The sole cytotoxic effect of evodiamine on SGC-7901 cells and human peripheral blood mononuclear cells (PBMCs) was observed by MTT assay. After the cells were respectively intervened with single evodiamine or evodiamine combined with z-VAD-fmk, the gene expressions of mTOR, p70S6K and 4EBP1 were analyzed by real-time PCR, and the protein expressions of mTOR and p-mTOR were detected by western blot. The result showed that evodiamine inhibited the apoptosis of SGC-7901 cells in a time-dependent manner, with no cytotoxic effect on human PBMCs. After the respective intervention with single evodiamine or evodiamine combined with z-VAD-fmk, the cells became round and floated in medium. Compared with the control group, both treatment methods can inhibit mTOR, 4E-BP1 and p70S6K gene expressions, with significant differences. Compared with single evodiamine, evodiamine combined with z-VAD-fmk showed a higher inhibitory rate in gene expression. According to the Western Blot result, evodiamine can inhibit the protein expressions of mTOR and p-mTOR regardless of the combination with z-VAD-fmk, with a higher inhibitory rate after z-VAD-fmk blocked caspase. In conclusion, evodiamine may promote the apoptosis of SGC-7901 cells through mTOR signal pathway.