Abstract The main feature of insomnia is difficulty in starting or maintaining sleep. 5-HT1A receptor and 5-HT2A receptor are two subtypes of the classic central 5-HT neurotransmitter closely related to sleep and wakefulness. To observe the effects of Evodia rutaecarpa acupoint sticking therapy on the mRNA expressions of 5-HT1A and 5-HT2A in the hypothalamus, brainstem, and hippocampus of insomnia rats induced by para-chlorophenylalanine (PCPA). Ten rats were randomly selected as the normal group, and 80 PCPA insomnia model rats were randomly divided into eight groups, including a model group, a positive control group (diazepam group), a low-dose E. rutaecarpa acupoint sticking therapy group (Yongquan acupoint group and Shenque acupoint group; the same applied below), a middle-dose E. rutaecarpa acupoint sticking therapy group, and a high-dose E. rutaecarpa acupoint sticking therapy group. The normal group and the model group did not receive treatment. The positive control group was given diazepam through intragastric administration, and the three-dose E. rutaecarpa acupoint sticking therapy groups were divided into the Yongquan acupoint group and the Shenque acupoint group. After seven days of administration, the rat hypothalamus, brainstem, and hippocampus tissues were taken, and a real-time polymerase chain reaction method was used to detect the mRNA expressions of 5-HT1A and 5-HT2A. E.rutaecarpa acupoint sticking therapy significantly upregulated 5-HT1A mRNA expression and downregulated 5-HT2A mRNA expression in rats with insomnia caused by PCPA. No significant differences were found in the expression between the two acupoints and the expression among the three brain tissues. E. rutaecarpa acupoint sticking therapy can improve insomnia. The mechanism may be related to the upregulation of 5-HT1A mRNA expression and the downregulation of 5-HT2A mRNA expression in brain tissue.

SUMMARY: The myodural bridge (MDB) is confirmed that connecting the most of suboccipital muscles to the cervical dura mater through the posterior intervertebral spaces and widely exists in mammals and birds. In order to reveal whether the MDB is universally existing in amniota of vertebrates, we explored the existence and the morphological features of the MDB in the Trachemys scripta elegans. Twenty fresh red-eared slider specimens were observed by the gross anatomy dissection and histological analysis. In the results, three kind of muscles in the postoccipital region of the red-eared slider were found. The rectus capitis dorsum minor muscle originated from the posterior margin of the occiput (C0) and terminated at the spinous process of the atlas (C1). The transversospinales muscle was attached to the vertebral arch and the postzygapophysis of the atlas and extended to the spinous process of the axis (C2). The C2-C3 intertransversales muscle were extended from the postzygapophysis of C2 and the one of C3. The three muscles covered the dorsal interspaces among C0-C3, and meantime they were closely connected with dense connective tissues, which filled in these interspaces. Each of these thick dense connective tissue membranes sent off several short and strong fibrous bundles ventrally to merge with the cervical spinal dura mater. Furthermore the connective tissues connecting these muscles with cervical spinal dura mater directly were revealed under the microscopy and they consisted of parallel and intensive collagen fibers with orientation from dorsal to ventral. In conclusion, this study for the first time demonstrated the existence of the MDB in the testudines, in all of the dorsal atlantooccipital, atlantoaxial and C2-C3 intervertebral spaces. Based on our results and comparative anatomical evidences in recent year, it could be inferred that the MDB might be its highly conserved structure in the evolution of amniota.

RESUMEN: Se confirma que el puente miodural (PMD) conecta la mayoría de los músculos suboccipitales con la duramadre cervical a través de los espacios intervertebrales posteriores y existe ampliamente en mamíferos y aves. Para revelar si el MDB existe universalmente en la amniota de vertebrados, exploramos la existencia y las características morfológicas del PMD en Trachemys scripta elegans. Veinte muestras se observaron mediante disección anatómica y análisis histológico. En los resultados, se encontraron tres tipos de músculos en la región occipital. El músculo recto capitis dorsum minor se originó en el margen posterior del occipital (C0) y terminó en el proceso espinoso del atlas (C1). El músculo transverso espinal se unió al arco vertebral y el proceso del atlas y se extendió al proceso espinoso del axis (C2). El músculo intertransversario C2-C3 se extendió entre los procesos transversos de C2 y el de C3. Los tres músculos cubrían los espacios intermedios dorsales entre C0-C3 y, mientras tanto, estaban estrechamente conectados con tejidos conectivos densos, que rellenaban estos espacios. Cada una de estas membranas densas de tejido conectivo envían varios haces fibrosos cortos y fuertes ventralmente para fusionarse con la duramadre espinal cervical. Además, los tejidos conectivos que conectan estos músculos con la duramadre cervical y espinal se revelaron directamente bajo microscopía y consistían en intensas fibras de colágeno, paralelas, con orientación desde dorsal a ventral. En conclusión, este estudio demostró por primera vez la existencia del PMD en los estudios de prueba, en todos los espacios dorsales atlantooccipital, atlantoaxial e intervertebral C2-C3. Sobre la base de nuestros resultados y las evidencias anatómicas comparativas de los últimos años, se podría inferir que el PMD podría ser una estructura altamente conservada en la evolución de la amniota.

Abstract The use of antimicrobial agents is an efficient method to prevent dental caries. Also, nanometric antibacterial agents with wide antibacterial spectrum and strong antibacterial effects can be applied for prevention of dental caries. Objectives: The aim of this study was to evaluate the inhibitory effect of reduced graphene oxide-silver nanoparticles (rGO/Ag) composite on the progression of artificial enamel caries in a Streptococcus mutans biofilm model. Material and Methods: Enamel specimens from bovine incisors were divided into eight treatment groups (n = 13), as follows: group 1 was inoculated with S. mutans grown in Brain Heart Infusion containing 1% sucrose (1% BHIS), as negative control; groups 2-4 were inoculated with S. mutans grown in the presence of different rGO/Ag concentrations (0.08, 0.12, 0.16 mg/mL) + 1% BHIS; group 5-7 were inoculated with S. mutans grown in the presence of different agents (0.16 mg/mL reduced graphene oxide, 0.16 mg/mL silver nanoparticles, 10 ppm NaF) + 1% BHIS; group 8 was mixed with 1% BHIS, without inoculation. Artificial enamel carious lesions were produced by S. mutans biofilm model for 7 days. Confocal laser scanning microscopy and atomic force microscopy were used to analyze roughness and morphology of the enamel surface. Polarized light microscopy and confocal laser scanning microscopy were employed to measure the lesion depth and the relative optical density (ROD) of the demineralized layer. Results: Compared with the control groups, the rGO/Ag groups showed: (a) reduced enamel surface roughness; (b) much smoother and less eroded surfaces; (c) shallower lesion depth and less mineral loss. Conclusion: As a novel composite material, rGO/Ag can be a promising antibacterial agent for caries prevention.

ABSTRACT The enzyme 3-hydroxy-3-methylglutaryl-CoA reductase (HMGR; EC1.1.1.34) catalyzes the first committed step of isoprenoids biosynthesis in Mevalonate (MVA) pathway. Here we report for the first time the cloning and characterization of a full-length cDNA encoding HMGR from Fritillaria cirrhosa (FcHMGR), a bulbous medicinal plant. The full-length cDNA of FcHMGR was 2072 base pair (bp), containing a 1680-bp open reading frame. Bioinformatical analyses revealed that FcHMGR had HMG CoA-binding domains and two NADPH binding domains, which are required for HMGR activity. Quantitative real-time PCR (qRT-PCR) analysis revealed that FcHMGR expressed high in mature bulbs. A truncated version of FcHMGR protein lacking the N-terminal 249-bp GC rich area was expressed in Escherichia coli. The crude cell lysate containing the recombinant protein showed a better HMGR activity than the control and the relative enzyme activity was calculated to be 1.62 U/mg. The cloning, characterization and functional analysis of FcHMGR gene allowed us to further understand the role of FcHMGR involved in steroidal alkaloid biosynthetic pathway in F. cirrhosa at the molecular level.

RESUMO Objetivo: Analisar se a associação entre fragilidade óssea e risco de fratura depende do nível de trauma. Método: Todos os participantes, juntamente com suas mães, foram submetidos à DEXA e medições corporais. Os participantes responderam um questionário de autorrelato sobre atividades físicas e descrição de como o trauma ocorreu. Os resultados do questionário foram associados à DEXA realizada na primeira visita do estudo. Resultados: Um total de 374 crianças com DEXA disponível e acompanhamento completo de 5 anos foi incluído na análise final. Das 374 crianças, 53 (14,2%) tiveram uma fratura e 11 (20,7%) tiveram mais de uma fratura. Com base na classificação de Landin modificada, foi determinado o nível de trauma. Das 53 (14,2%) crianças que tiveram uma fratura, 39 (73,6%) foram classificadas, sendo 19 (48,7%) com trauma leve, 16 (41%) com trauma moderado e quatro (10,2%) com trauma grave. O nível de trauma não pôde ser atribuído a 14 (26,4%) crianças, devido a informações limitadas. As crianças sem fraturas apresentaram valores significantemente mais altos em todos os parâmetros ósseos, em comparação com os que tinham fraturas causadas por trauma leve. Conclusão: Os indivíduos com fraturas por trauma leve apresentaram relação inversamente proporcional entre os parâmetros fragilidade óssea e o risco de fratura em comparação com indivíduos sem fratura. Nível de Evidência IV, Série de Casos .

ABSTRACT Objective: To analyze whether association between bone fragility and risk of fracture depends on the trauma level. Method: All participants along with their mothers underwent DXA scan and body measurements. The subjects answered a self-report questionnaire about their physical activities and the precipitating causes. The questionnaire results were associated with DXA performed at the baseline visit. Results: A total 374 children with available DXA scan and complete follow-up of 5 years were included in the final analysis. Of the 374 children, 53 (14.2%) had one fracture, and 11 (20.7%) had more than one fracture. Based on the modified Landin classification, the trauma level was determined. Of the 53 (14.2%) children who had one fracture, 39 (73.6%) were classified, namely 19 (48.7%) with mild trauma, 16 (41%) with moderate trauma and four (10.2%) with severe trauma. Trauma level could not be assigned to 14 (26.4%) children due to limited information. Children without fractures had significantly higher values in all bone parameters compared to those with fractures caused by mild trauma. Conclusion: Subjects with mild trauma fractures had an inversely proportional ratio between bone fragility parameters and fracture risk compared with subjects without fractures. Level of Evidence IV, Case Series.

摘要 目的 旨在确定上海地区肺结核病患者的超额死亡率。 方法 参与者是曾于2004 年 1 月 1 日至 2008 年 12 月 31 日在四家肺结核病诊所其中一家登记过的、上海市 4 个区（共 19 个区）的当地居民。我们收集了最近的肺结核病诊断的基线数据，并对 2014 年 3 月至 5 月的死亡率进行了评估。我们计算出所有参与者和子组的标准化死亡率和病死率，并采用单变量和多变量 Cox 回归模型量化各种原因与肺结核导致的共病症与死亡率之间的关联。 结果 我们登记了病患中 4569 名受试者。总体而言，各种原因导致的死亡患者的标准化死亡率为 5.2 （95% 置信区间：4.8-5.6）。男性的标准化死亡率高于女性 (6.1 比 3.0)。调整年龄和性别后，以前接受过治疗的患者（风险比：1.26，95% 置信区间：1.08-1.49）和痰涂片试验呈阳性的患者（风险比：1.55，95% 置信区间：1.35-1.78）中各种原因导致的死亡风险比明显更高。以前接受过治疗的患者（风险比率：1.88，95% 置信区间：1.24-2.86）和痰涂片试验呈阳性的患者（风险比率：3.16，95% 置信区间：2.06-4.87）中肺结核导致的死亡风险也明显更高。 结论 上海地区肺结核病患者的死亡率风险增加。及早的诊断和更加警觉的随访诊查可能有助于降低肺结核患者的死亡率。

Résumé Objectif Déterminer la surmortalité dans une cohorte de personnes atteintes de tuberculose à Shanghai. Méthodes Les participants étaient des habitants de 4 districts (sur 19) de Shanghai, enregistrés dans une des quatre cliniques de la tuberculose entre le 1er janvier 2004 et le 31 décembre 2008. Des données de référence ont été recueillies lors du diagnostic le plus récent de tuberculose et la mortalité a été évaluée entre mars et mai 2014. Nous avons calculé les rapports standardisés de mortalité (RSM) et les taux de létalité pour tous les participants ainsi que pour des sous-groupes. Des modèles de régression univariée et multivariée de Cox ont été utilisés pour quantifier les associations entre les comorbidités et la mortalité, toutes causes confondues et due à la tuberculose. Résultats Nous avons enregistré 4569 sujets dans la cohorte. Globalement, la cohorte avait un RSM pour les décès toutes causes confondues de 5,2 (intervalle de confiance (IC) de 95%: 4,8-5,6). Les hommes affichaient un RSM plus élevé que les femmes (6,1 contre 3,0). Après ajustement en fonction de l'âge et du sexe, les ratios de risque (RR) de décès toutes causes confondues étaient nettement plus élevés chez les personnes ayant précédemment reçu un traitement (RR: 1,26; IC 95%: 1,08-1,49) et les personnes à frottis d'expectoration positif (RR: 1,55; IC 95%: 1,35-1,78). Le risque de décès dû à la tuberculose était également beaucoup plus important pour les personnes ayant précédemment reçu un traitement (RR: 1,88; IC 95%: 1,24-2,86) et les personnes à frottis positif (RR: 3,16; IC 95%: 2,06-4,87). Conclusion Les personnes atteintes de tuberculose à Shanghai présentent un risque accru de mortalité. Un diagnostic plus précoce et un suivi plus attentif pourraient permettre de réduire la mortalité dans ce groupe.

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Abstract Objective To determine excess mortality in a cohort of people with tuberculosis in Shanghai. Methods Participants were local residents in 4 (of 19) districts in Shanghai, registered in one of four tuberculosis clinics between January 1, 2004 and December 31, 2008. Baseline data were collected at the most recent diagnosis of tuberculosis and mortality was assessed between March and May of 2014. We calculated standardized mortality ratios (SMR) and case-fatality rates for all participants and for subgroups. Univariate and multivariate Cox regression models were used to quantify associations between co-morbidities and mortality from all causes and from tuberculosis. Findings We registered 4569 subjects in the cohort. Overall, the cohort had an SMR for deaths from all causes of 5.2 (95% confidence interval, CI: 4.8-5.6). Males had a higher SMR than females (6.1 versus 3.0). After adjustment for age and sex, hazard ratios (HR) for deaths from all causes were significantly greater in previously treated people (HR: 1.26; 95% CI: 1.08-1.49) and sputum smear-test positive people (HR: 1.55; 95% CI: 1.35-1.78). The risk of death from tuberculosis was also significantly greater for previously treated people (HR: 1.88; 95% CI: 1.24-2.86) and smear positive people (HR: 3.16; 95% CI: 2.06-4.87). Conclusion People with tuberculosis in Shanghai have an increased risk of mortality. Earlier diagnosis and more vigilant follow-up may help to reduce mortality in this group.

ملخص الغرض تحديد الزيادة في حالات الوفاة في مجموعة من المصابين بالسل في شانغهاي. الطريقة كان المشاركون من السكان المحليين في 4 أحياء (من بين 19 حيًا) في شنغهاي، وقد تم تسجيلهم في واحدة من أربع عيادات لعلاج السل في الفترة بين 1 يناير/كانون الثاني 2004 و31 ديسمبر/كانون الأول 2008. وقد تم جمع البيانات الأساسية على مستوى أحدث الحالات التي تم تشخيصها كحالات الإصابة بالسل، كما تم تقدير الوفيات في الفترة بين شهري مارس/آذار ومايو/أيار 2014. وحسبنا نسب الوفيات القياسية (SMR) ومعدلات الوفيات بين الحالات المصابة بالنظر إلى جميع المشاركين والمجموعات الفرعية.وتم استخدام نماذج انحدار كوكس ذات المتغير الأحادي والمتعددة المتغيرات لقياس مدى الارتباط بين الإصابة بأكثر من مرض في نفس الوقت وحالات الوفاة الناتجة عن جميع الأسباب والناتجة عن مرض السل. النتائج سجلنا 4569 من الحالات محل الدراسة في المجموعة المشار إليها.وبوجه عام، بلغت نسبة الوفيات القياسية الناتجة عن جميع الأسباب في هذه المجموعة 5.2 (بنسبة أرجحية مقدارها 95%: 4.8-5.6). وارتفعت نسبة الوفيات القياسية بين الرجال مقارنةً بالنساء (6.1 مقابل 3.0).وبعد تعديل عامل العمر والجنس، كانت نسب خطورة (HR) التعرض للوفاة الناتجة عن جميع الأسباب أكثر ارتفاعًا بشكل ملحوظ لدى الأشخاص الذين سبقت معالجتهم (نسبة خطورة: 1.26؛ بنسبة أرجحية مقدارها 95%: 1.08- 1.49) ولدى الأشخاص الذين كانت نتائج اختبار مسحة البلغم الخاصة بهم إيجابية (HR: 1.55؛ بنسبة أرجحية مقدارها 95%: 1.35-1.78). وكان خطر الوفاة الناتجة عن السل أكثر ارتفاعًا أيضًا بشكل ملحوظ لدى الأشخاص الذين سبقت معالجتهم (نسبة خطورة: 1.88؛ بنسبة أرجحية مقدارها 95%: 1.24- 2.86) ولدى الأشخاص الذين كانت نتائج المسحة الخاصة بهم إيجابية (نسبة خطورة: 3.16؛ بنسبة أرجحية مقدارها 95%: 2.06-4.87). الاستنتاج يعاني المصابون بالسل في شانغهاي من زيادة خطر التعرض للوفاة. وقد يساعد التشخيص المبكر للمرض والمزيد من المتابعة الطبية الدقيقة في تقليل معدل الوفيات في هذه المجموعة.

Resumen Objetivo Determinar el exceso de mortalidad en una cohorte de personas con tuberculosis en Shanghái. Métodos Los participantes eran residentes locales de 4 (de 19) distritos de Shanghái, registrados en uno de los cuatro consultorios antituberculosos entre el 1 de enero de 2004 y el 31 de diciembre de 2008. Se recogieron datos de referencia de los diagnósticos de tuberculosis más recientes y se evaluó la mortalidad entre marzo y mayo de 2014. Se calcularon las tasas de mortalidad estandarizadas (SMR) y la tasa de letalidad de todos los participantes y por subgrupos. Se utilizaron modelos de regresión de Cox univariables y multivariables para cuantificar las asociaciones entre la comorbilidad y la mortalidad de todas las causas de tuberculosis. Resultados Se registraron 4.569 sujetos en la cohorte. En general, la cohorte tenía una SMR para muertes de todas las causas de 5,2 (intervalo de confianza del 95%, IC: 4,8-5,6). Los hombres tenían una SMR mayor que las mujeres (6,1 frente a 3,0). Tras ajustarlos de acuerdo a edad y sexo, los cocientes de riesgo (HR) para las muertes de todas las causas eran significantemente mayores en personas tratadas con anterioridad (HR: 1,26 (IC del 95%: 1,08-1,49) y personas con pruebas de esputo positivas (HR: 1,55 (IC del 95%: 1,35-1,78). El riesgo de muerte por tuberculosis también era significativamente mayor en personas tratadas con anterioridad (HR: 1,88 (IC del 95%: 1,24-2,86) y personas con pruebas de esputo positivas (HR: 3,16 (IC del 95%: 2,06-4,87). Conclusión Las personas con tuberculosis en Shanghái tienen un mayor riesgo de mortalidad. Unos diagnósticos más tempranos y un seguimiento más atento podrían ayudar a reducir la mortalidad de este grupo.

The JAK2/STAT3 signal pathway is an important component of survivor activating factor enhancement (SAFE) pathway. The objective of the present study was to determine whether the JAK2/STAT3 signaling pathway participates in hydrogen sulfide (H2S) postconditioning, protecting isolated rat hearts from ischemic-reperfusion injury. Male Sprague-Dawley rats (230-270 g) were divided into 6 groups (N = 14 per group): time-matched perfusion (Sham) group, ischemia/reperfusion (I/R) group, NaHS postconditioning group, NaHS with AG-490 group, AG-490 (5 µM) group, and dimethyl sulfoxide (DMSO; <0.2%) group. Langendorff-perfused rat hearts, with the exception of the Sham group, were subjected to 30 min of ischemia followed by 90 min of reperfusion after 20 min of equilibrium. Heart rate, left ventricular developed pressure (LVDP), left ventricular end-diastolic pressure (LVEDP), and the maximum rate of increase or decrease of left ventricular pressure (± dp/dt max) were recorded. Infarct size was determined using triphenyltetrazolium chloride (TTC) staining. Myocardial TUNEL staining was used as the in situ cell death detection method and the percentage of TUNEL-positive nuclei to all nuclei counted was used as the apoptotic index. The expression of STAT3, bcl-2 and bax was determined by Western blotting. After reperfusion, compared to the I/R group, H2S significantly improved functional recovery and decreased infarct size (23.3 ± 3.8 vs 41.2 ± 4.7%, P < 0.05) and apoptotic index (22.1 ± 3.6 vs 43.0 ± 4.8%, P < 0.05). However, H2S-mediated protection was abolished by AG-490, the JAK2 inhibitor. In conclusion, H2S postconditioning effectively protects isolated I/R rat hearts via activation of the JAK2/STAT3 signaling pathway.