The "5Ts" screening tool: Enhancements and threshold values for effective TMD identification.

OBJECTIVE: This study aimed to enhance the quintessential "five temporomandibular disorder (TMD) symptoms" (5Ts) screener by incorporating frequency options and distinguishing between TMJ and muscle pain. The diagnostic accuracy along with cut-off points for the effective identification of TMDs was also established. METHODS: Participants, aged ≥18 years, were recruited from a university-based hospital. After completing surveys encompassing demographic data and the enhanced 5Ts (with frequency options [5Ts-F] and differentiation of TMJ/muscle pain [6Ts-F]), protocolized interviews and clinical examinations were performed following DC/TMD. The diagnostic accuracy and best cut-off points were determined with the area under the receiver operating characteristic curves (AUCs). RESULTS: 324 participants were recruited (mean age 30.0 ± 11.4 years). Among these, 86.4% had TMDs. 5Ts exhibited high diagnostic accuracy for detecting all TMDs (AUC = 0.92) with sensitivity/specificity values of 83.9%/88.6%. Both 5Ts-F and 6Ts-F had slightly better accuracy (AUCs = 0.95/0.96), comparable sensitivity, and superior specificity (97.7%) compared to 5Ts. The best cut-off points were 1.5 for 5Ts and 2.5 for 5Ts-F/6Ts-F. CONCLUSIONS: Although all three TMD screeners presented high diagnostic accuracy, 5Ts-F/6Ts-F had notably improved specificity. 5Ts scores of >1.5 and 5Ts-F/6Ts-F scores of >2.5 are to be applied for screening the presence of TMDs.

Greater efficacy of a combination of conservative therapies for thumb base OA in individuals with lower radial subluxation - a pre-planned subgroup analysis of the COMBO trial.

OBJECTIVE: To investigate heterogeneous effects of a combination of conservative therapies compared with an education comparator for thumb base (TB) osteoarthritis (OA) according to clinically relevant characteristics. METHODS: Pre-planned subgroup analysis of the COMBO trial (n = 204) which compared a combination of education on self-management and ergonomic principles, a prefabricated neoprene splint, hand exercises, and diclofenac sodium gel, with education alone for radiographic and symptomatic TB OA. Primary outcomes were change in pain (visual analogue scale [VAS], 0-100 mm) and hand function (Functional Index for Hand Osteoarthritis questionnaire, 0-30) from baseline to week-6. Other outcomes were grip and tip-pinch strength and patient's global assessment (PGA) (VAS, 0-100 mm). Possible treatment effect modifiers were the presence of interphalangeal joint pain, erosive hand OA, radiographic thumb carpometacarpal joint subluxation (higher vs equal or lower than the sample mean), and baseline radiographic OA severity (Kellgren Lawrence grade). Linear regression models were fitted, adding interaction terms for each subgroup of interest. RESULTS: The treatment effects of the combined intervention at 6 weeks were greater in participants with lower joint subluxation compared with those with greater subluxation (pain -11.6 [95%CI -22.2, -9.9] and 2.6 [-5.5, 10.7], respectively, difference between the subluxation groups 14.2 units (95% CI 2.3, 26.1), p-value 0.02; and PGA -14.0 [-22.4, -5.5] and 1.5 [-6.2, 9.3), respectively, difference between the subluxation groups 15.5 units (95% CI 4.2, 26.8), p-value 0.03). There was no statistically significant heterogeneity for the other subgroups. CONCLUSION: A combination of conservative therapies may provide greater benefits over 6 weeks in individuals with lower joint subluxation, although the clinical relevance is uncertain given the wide confidence intervals. Treatment strategies may need to be customized for those with greater joint subluxation. TRIAL REGISTRATION NUMBER: ACTRN 12616000353493.

Ecdysis triggering hormone is essential for larva-pupa-adult transformation in Leptinotarsa decemlineata.

In Drosophila melanogaster, ecdysis triggering hormone (ETH) is the key factor triggering ecdysis behaviour and promoting trachea clearance. However, whether ETH plays the dual roles in non-dipteran insects is unknown. In this survey, we found that Ldeth mRNA levels were positively correlated with circulating 20-hydroxyecdysone (20E) titers in Leptinotarsa decemlineata. Ingestion of an ecdysteroid agonist halofenozide or 20E stimulated the transcription of Ldeth, whereas RNA interference (RNAi) of ecdysteroidogenesis (LdPTTH or LdSHD) or 20E signalling (LdEcR, LdUSP or LdFTZ-F1) genes inhibited the expression, indicating ETH acts downstream of 20E. RNAi of Ldeth at the final instar stage impaired pupation. More than 80% of the Ldeth-depleted beetles remained as prepupae, completely wrapped in the old larval cuticles. These prepupae became withered, dried and darkened gradually, and finally died in soil. The remaining Ldeth hypomorphs pupated and emerged as abnormal adults, bearing smaller and wrinkle elytrum and hindwing. Moreover, the tracheae in the Ldeth hypomorphs were full of liquid. We accordingly proposed that the failure of trachea clearance disenabled air-swallowing after pupa-adult ecdysis and impacted wing expansion. Our results suggest that ETH plays the dual roles, initiation of ecdysis and motivation of trachea clearance, in a coleopteran.

Association of weather factors with the risk of pain exacerbations in people with hip osteoarthritis.

Objectives: Our objective was to evaluate the association of weather factors with the risk of pain exacerbations in people with symptomatic hip osteoarthritis (OA). Method: Eligible participants with symptomatic hip OA were instructed to log on to the study website and complete questionnaires every 10 days and additionally whenever they considered they were experiencing a pain exacerbation (case period) during the 90 day follow-up. Pain exacerbation was defined as an increase of two points in pain intensity on an 11-point numeric rating scale (0-10) during the follow-up compared with baseline. Each case period was anchored to four control periods within a 35 day interval using a time-stratified approach. Weather data were obtained for both periods from the publicly available meteorological database of the Australian Bureau of Meteorology. We examined the association of weather factors across 72 h before the index date with the risk of pain exacerbation, using conditional logistic regression. Results: Among 252 participants recruited, 129 participants had at least one episode of pain exacerbation and were included in the analysis. A significant dose-response relationship was found between average daily temperature variation in the prior 72 h and risk of pain exacerbations (p = 0.04 for linear trend). There was no significant association between maximum daily temperature, minimum daily temperature, relative humidity, precipitation, or barometric pressure and hip pain exacerbations. Conclusion: The overall results suggest that only daily temperature variation among different weather factors was associated with hip pain exacerbations in people with symptomatic hip OA.

Estimation of effective dose of propofol mono-sedation for successful insertion of upper gastrointestinal endoscope in healthy, non-obese Chinese adults.

WHAT IS KNOWN AND OBJECTIVE: Propofol is effective in sedation for upper gastrointestinal (UGI) endoscopy. However, the optimum dose is ill-defined. This study aimed to estimate the effective dose of propofol mono-sedation for successful endoscope insertion in healthy, non-obese Chinese adults undergoing single UGI endoscopy. METHODS: Twenty-six adult patients undergoing elective single UGI endoscopy were enrolled in this study. A modified Dixon's up-and-down method was utilized to assess the effective dose of propofol for successful endoscope insertion. The initial dose of propofol administered, 1.6 mg/kg, was adjusted with 0.1 mg/kg as a step size. The patient's responses to endoscope insertion were classified as either 'movement' or 'no movement'. When patient's responses were changed from 'movement' to 'no movement' or from 'no movement' to 'movement', a crossover was defined. After eight crossovers had been obtained, patient recruitment was stopped. The mean of midpoints of all crossovers obtained by the modified Dixon's up-and-down method in all 26 patients was defined as calculated median effective dose (ED50 ) of propofol for successful endoscope insertion. Furthermore, probit regression analysis was used to determine the dose of propofol where 50% (ED50 ) and 95% (ED95 ) of endoscope insertion attempts were successful. RESULTS: The calculated ED50 of propofol for successful endoscope insertion was 1.89 ± 0.12 mg/kg. The probit regression analysis showed that ED50 and ED95 of propofol for successful endoscope insertion were 1.90 mg/kg (95% CI, 1.78-2.10 mg/kg) and 2.15 mg/kg (95% CI, 2.01-3.56 mg/kg), respectively. No any patient had hypoxaemia and gag reflex during the UGI endoscopy with propofol mono-sedation. WHAT IS NEW AND CONCLUSION: In healthy, non-obese Chinese adults, propofol mono-sedation can provide excellent conditions of UGI endoscopy and the estimated ED50 of propofol for successful endoscope insertion is 1.89 ± 0.12 mg/kg.

[Application of neuroendoscopy combined with fluorescence angiography in anterior circulation aneurysm clipping].

Objective: To explore the value of neuroendoscopy combined with fluorescence angiography in anterior circulation aneurysm clipping. Methods: A total of 15 patients with anterior circulation aneurysm from Department of Neurosurgery, Zhongnan Hospital of Wuhan University between October 2018 and January 2019 were enrolled. Neuroendoscopy combined with indocyanine green fluorescence angiography (ICGA) was used to determine the shape of the aneurysm, the specific location of the aneurysm neck and its relationship with the aneurysm-bearing artery during anterior circulation aneurysm clipping. Meanwhile, Neuroendoscopy combined with ICGA can be employed to observe whether there was stenosis and incomplete clamping of the aneurysm-bearing artery after clipping the aneurysm, and whether there was misclamping of the perforating branches hidden under the posterior wall of the aneurysm. Results: The success rate of aneurysm clipping in 15 cases was 15/15. After aneurysm clipping, ICGA and neuroendoscopy were performed. The residual aneurysm neck was detected in 3 cases, and the position of aneurysm clip was adjusted or aneurysm clips were added. In one case, the anterior choroidal aneurysm was found to be mistakenly clipped. After adjusting the aneurysm clip, ICGA and neuroendoscopy showed that the anterior choroidal artery was normal. In another case, the A1 segment aneurysm was clipped. ICGA and neuroendoscopy found that the perforating branch blood vessels were mistakenly clipped. After the adjustment of the aneurysm clip, the blood vessels recovered their patency. There were no surgical-related deaths, disability and coma cases in the study. Conclusions: During aneurysm clipping, neuroendoscopy combined with ICGA can reduce cerebral vasospasm, decrease the misclipping rate of perforation of blood vessels, and avoid residual neck of aneurysm, stenosis or occlusion of aneurysm-bearing artery by using neuroendoscopy to observe whether misclipping of the perforating branch vessels exist and whether the aneurysm is clipped. Therefore, it can reduce postoperative complications.

[Analysis of incidence and risk factors of neonatal ventilator associated pneumonia in a hospital in Hunan Province, 2016-2018].

Objective: To investigate the incidence and risk factors of ventilator-associated pneumonia (VAP) in neonatal intensive care unit (NICU), and to provide evidence for the prevention and control of VAP. Methods: A total of 1 872 neonates, who were admitted into NICU of Hunan Provincial Maternal and Child Health Hospital and subjected to mechanical ventilation from October 2016 to June 2018, were enrolled in the study. The neonates who met the diagnostic criteria of VAP were selected as the case group, and those who were treated with ventilator for 48 hours at the same time were regarded as the control group. Multivariate logistic regression model was used to analyze the related factors of VAP. Results: Of the 1 872 neonates who underwent the mechanical ventilation, the VAP occurred in 160 cases with the incidence rate of 8.5% (160 cases). The 227 specimens were collected. Gram-positive bacteria (n=116, 51.1%) were the main pathogens. The main pathogens were Staphylococcus epidermidis, Enterococcus faecalis, Acinetobacter baumannii. By Chi-square test, birth weight, birth age, Apgar score, duration of ventilator, and whether newborn mothers with pregnancy hypertension were influencing factors. The result of logistic regression analysis showed that compared with no pregnancy included hypertension, the first aid measure at birth was initial resuscitation, and the MV time ≤ 5 days, the risk factors of ventilator-associated pneumonia in neonates included: their mothers with hypertensive disorders complicating pregnancy, using of tracheal intubation and ventilator time more than 5 days. Conclusion: The incidence of VAP in neonates receiving continuous MV therapy in neonatal intensive care unit is higher. Gram-positive bacteria are the main pathogens. VAP in neonates is related to whether newborn mothers with pregnancy hypertension, MV duration and tracheal intubation.

Functional characterization of ultraspiracle in Leptinotarsa decemlineata using RNA interference assay.

A heterodimer of ultraspiracle (USP) and ecdysone receptor (EcR) mediates 20-hydroxyecdysone (20E) signalling cascade to regulate insect moulting and metamorphosis. However, at least two questions remain to be addressed in terms of the molecular importance of USP in insect species. First, is USP involved in both regulation of ecdysteroidogenesis and mediation of 20E signalling in non-drosophilid insects, as in Drosophila melanogaster? Second, does USP play any role in larval metamorphosis except as the partner of heterodimeric receptor to activate the downstream 20E signalling genes? In this paper, we found that RNA interference (RNAi) of LdUSP in the final (fourth) instar larvae reduced the messenger RNA levels of four ecdysteroidogenesis genes (Ldspo, Ldphm, Lddib and Ldsad) and 20E titre, and repressed the expression of five 20E signal genes (EcRA, HR3, HR4, E74 and E75) in Leptinotarsa decemlineata. The LdUSP RNAi larvae remained as prepupae, with developing antennae, legs and discs of forewings and hindwings. Dietary supplement with 20E restored the expression of the five 20E signal genes, but only partially alleviated the decreased pupation rate in LdUSP RNAi beetles. Knockdown of LdUSP at the penultimate (third) instar larvae did not affect third-fourth instar moulting. However, silencing LdUSP caused similar but less severe impairments on pupation. Accordingly, we propose that USP is undoubtedly necessary for ecdysteroidogenesis, for mediation of 20E signalling and for initiation of metamorphosis in L. decemlineata.

Silencing chitin deacetylase 2 impairs larval-pupal and pupal-adult molts in Leptinotarsa decemlineata.

Insect chitin deacetylases (CDAs) are carbohydrate esterases that catalyze N-deacetylation of chitin to generate chitosan, a process essential for chitin organization and compactness during the formation of extracellular chitinous structure. Here we identified two CDA2 splice variants (LdCDA2a and LdCDA2b) in Leptinotarsa decemlineata. Both splices were abundantly expressed in larval foregut, rectum, and epidermis; their levels peaked immediately before ecdysis within each instar. In vivo results revealed that the two isoforms transcriptionally responded, positively and negatively respectively, to 20-hydroxyecdysone and juvenile hormone signaling pathways. RNA interference (RNAi)-aided knockdown of the two LdCDA2 variants (hereafter LdCDA2) or LdCDA2b, rather than LdCDA2a, resulted in three negative effects. First, foliage consumption was significantly reduced, larval developing period was lengthened, and larval growth was retarded. Second, chitin contents were reduced, whereas glucose, trehalose, and glycogen contents were increased in the LdCDA2 and LdCDA2b RNAi larvae. Third, approximately 20% of LdCDA2 and LdCDA2b RNAi larvae were trapped within the exuviae and finally died. About 60% of the abnormal pupae died as pharate adults. Around 20% of the RNAi pupae emerged as deformed adults, with small size and wrinkled wings. These adults eventually died within 1 week after molting. Our results reveal that knockdown of CDA2 affects chitin accumulation. Consequently, LdCDA2 may be a potential target for control of L. decemlineata larvae.

Isoform specific roles of Broad-Complex in larval development in Leptinotarsa decemlineata.

Broad-Complex (BrC) is a downstream target of both 20-hydroxyecdysone and juvenile hormone signalling. BrC regulates morphogenetic changes between nymphal instars in hemimetabolans, whereas it controls pupal commitment, pupal morphogenesis and inhibits adult differentiation in holometabolans. Among five BrC cDNAs (Z1-Z4 and Z6) identified in the Colorado potato beetle, we found in this work that Z1, Z2 and Z6 were mainly expressed at the last (fourth) instar and prepupal stages, whereas the levels of Z3 and Z4 increased during the penultimate (third) instar stage, peaked at the last instar larval phase and gradually decreased at the prepupal and pupal periods. When knocking down all BrC isoforms by RNA interference (RNAi) at the penultimate instar stage, around 20% of the resultant larvae remained as moribund beetles. These moribund BrC RNAi larvae were completely or partially wrapped in old cuticle. Likewise, a portion of larvae treated for a single double-stranded RNA of Z3, Z4 or Z6 displayed a degree of similar aberrancies, increasing in the order of isoforms Z6 < Z3 < Z4. When silencing all BrC isoforms at the last instar period, most of the RNAi larvae did not normally pupate or emerge as adults. Separately silencing each of the five zinc finger domains revealed that approximately 70% of the Z1 RNAi larvae remained as prepupae, around 60% of the Z6 RNAi specimens formed aberrant prepupae or pupae and about 60% of the Z2 RNAi beetles became deformed pupae. After removal of the old exuviae, these deformed larvae in which either Z1, Z2 or Z6 was depleted possessed adult prothorax and mesothorax, developing antenna, mouthparts and wing discs. Moreover, less than 50% of the resultant pupae finally emerged as adults when either of Z1, Z2 or Z6 was knocked down. Therefore, our findings reveal, for the first time, that the two roles of BrC in insect groups (ie directing morphogenetic changes during juvenile development and regulating larval-pupal-adult metamorphosis) are played by different BrC isoforms in Leptinotarsa decemlineata.

Impairment of pupation by RNA interference-aided knockdown of Broad-Complex gene in Leptinotarsa decemlineata (Say).

Dietary delivery of bacterially expressed double-stranded RNA (dsRNA) has a great potential for management of Leptinotarsa decemlineata. An important first step is to discover possible RNA-interference (RNAi)-target genes effective against larvae, especially the old larvae. In the present paper, five putative Broad-Complex (BrC) cDNAs (Z1-Z4, and Z6) were identified in L. decemlineata. The expression of the five LdBrC isoforms was suppressed by juvenile hormone signaling, whereas the transcription was upregulated by 20-hydroxyecdysone signaling at the fourth (final) instar larval stage. Feeding of bacterially expressed dsBrC (derived from a common fragment of the five LdBrC variants) in the third- and fourth-instar larvae successfully knocked down the target mRNAs. For the fourth-instar LdBrC RNAi hypomorphs, they had a higher larval mortality compared with the controls. Moreover, most dsBrC-fed beetles did not pupate normally. After removal of the apolysed larval cuticle, a miniature adult was found. The adult head, compound eyes, prothorax, mesothorax, metathorax were found on the dorsal view. Distinct adult cuticle pigmentation was seen on the prothorax. The mouthparts, forelegs, midlegs, and hindlegs could be observed on the ventral view of the miniature adults. For the third-instar LdBrC RNAi specimens, around 20% moribund beetles remained as prepupae and finally died. Therefore, LdBrC is among the most attractive candidate genes for RNAi to control the fourth-instar larvae in L. decemlineata.

[Computed tomographic features of desmoplastic ameloblastoma of the jaw].

OBJECTIVE: To retrospectively analyze the clinical and CT imaging features of desmoplastic ameloblastomas (DA) in comparison with other types of ameloblastomas. METHODS: Patients diagnosed with ameloblastoma histopathologically in Peking University School and Hospital of Stomatology from July 2000 to August 2017 were reviewed to analyze the constituent ratios and characteristics of DA. CT imaging features of DA (28 cases) were investigated in comparison with consecutive cases of solid/multicystic and unicystic ameloblastomas. The following imaging perspectives were analyzed: the border, internal structure, three-dimensional shape, tooth, the periodontium, the cortex and the expansion of the tumors. CT values were measured in the region of interest for quantitative analysis. RESULTS: Among the 1 269 cases of ameloblastomas, 50 (3.9%) were DA, which showed predilections for males (74.0%). The average age of DA was about 43.9 years old, which was higher than the other two types. The anterior incisorcanine region (62.0%) and premolar region (30.0%) were most frequently affected. The incidence rate of DA in mandibule was 56.0% (28/50), which was slightly higher than that of maxilla (44.0%). The DA characteristically showed scallop border and honeycomblike or soapbubble internal structure with bone formation on CT. The mean ratios of height to mesiodistal and buccal-lingual to mesio-distal distances were 0.76 and 0.63, which were higher than the other two types. According to the degree of internal bone formation, three subtypes of DA could be observed: densely ossifying type (I), honeycomb/soap bubble type (II) and sparsely ossifying type (III). The means and standard deviations of CT values of DA were significantly higher than those of the other two types, which were (488.8±164.0) HU (type I), (171.7±102.8) HU (type II), (42.1±8.8) HU (type III). CONCLUSION: CT is helpful for diagnosis of DA, which shows as solid tumor with varying degrees of internal ossification.

Requirement of Leptinotarsa decemlineata gene within the 74EF puff for larval-pupal metamorphosis and appendage growth.

Two Drosophila melanogaster E-twenty-six domain transcription factor isoforms (E74A and E74B) act differentially at the start of the 20-hydroxyecdysone (20E) signalling cascade to regulate larval-pupal metamorphosis. In the present paper, we identified the two isoforms (LdE74A and LdE74B) in Leptinotarsa decemlineata. During the larval development stage, the mRNA transcript levels of the two LdE74 isoforms were correlated with circulating 20E titres. In vitro midgut culture and in vivo dietary supplementation with 20E revealed that the presence of 20E induced expression peaks of both LdE74A and LdE74B, with similar patterns observed for the two isoforms. Moreover, the mRNA transcript levels of both LdE74A and LdE74B isoforms were significantly downregulated in the L. decemlineata ecdysone receptor RNA interference (RNAi) specimens, but not in the LdE75 RNAi beetles. Ingestion of 20E reduced the larval fresh weights and shortened the larval development period, irrespective of knockdown of LdE74 or not. RNAi of LdE74 did not affect 20E-induced expression of the Ecdysone induced protein 75-hormone receptor 3-fushi tarazu factor 1 (E75-HR3-FTZ-F1) transcriptional cascade. Thus, it seems that LdE74 mediates 20E signalling independent of the E75-HR3-FTZ-F1 transcriptional cascade. Furthermore, silencing of both LdE74 isoforms caused failure of ecdysis. Most of the LdE74 RNAi beetles remained as prepupae. The LdE74 RNAi prepupae exhibited adult character-like forms underneath after removal of the apolysed larval cuticle. Their appendages such as antennae, legs and wings were shorter than those of control larvae. Only a few LdE74 RNAi larvae finally became deformed pupae, with shortened antennae and legs. Therefore, LdE74 is required for larval-pupal metamorphosis and appendage growth in L. decemlineata.

Physiological effects of anterior repositioning splint on temporomandibular joint disc displacement: a quantitative analysis.

Anterior repositioning splints (ARS) are used primarily for the management of temporomandibular joint (TMJ) anterior disc displacement with reduction (ADDwR). However, the exact physiological effects of ARS are still unclear. This study investigated the short and long-term effects of ARS on disc and condyle angles/positions by metric analysis. Twenty-two subjects diagnosed with ADDwR were recruited. Maxillary full-coverage ARS were fabricated, and MRI of TMJs was obtained before splint treatment, immediate post-insertion and 6 months after splint treatment. Disc-condyle relationship was determined by disc-condyle angle measurement. Disc and condyle positions were described as X-Y coordinates with the summit of glenoid fossa as the origin of the coordinates. Thirty-two TMJs were classified as ADDwR and 12 were normal. Upon ARS insertion, all TMJs with ADDwR got normal disc-condyle relationships. The condyles moved significantly forward and downward, while the discs moved significantly backward and upward. MRI at 6 months after treatment (without ARS insertion) indicated that only 40·6% (13/32) of the joints were maintained in the normal disc-condyle relationship. The majority of condyles returned to their pre-treatment positions, while the discs generally moved anteriorly again. The use of ARS resulted in forward and downward condyle movement, and a concurrent backward movement of the disc resulting in ideal spatial disc-condyle relationship. The stability of this relationship, however, could not be maintained in the majority of TMJs upon ARS removal. Findings explain the good short-term clinical outcomes with ARS and their relatively lower efficacy in the long term.

[Changes of productions of energy metabolism in masseter of rats induced by occlusal interference].

OBJECTIVE: To investigate the effect of occlusal interference on the energy metabolism of masticatory muscle by studying the changes of adenosine triphosphate (ATP), adenosine diphosphate (ADP), inosine monophosphate (IMP), phosphocreatine, creatine, lactate and pH level in masseter muscles of rats after occlusal interference. METHODS: Fifty male Sprague-Dawley rats were randomly assigned into experimental group (n=40) and control group (n=10). In experimental group, 0.4 mm thick metal crown was cemented to the upper right first molar of the rat, and maintained for 3, 7, 10, 14 d separately (n=10 for each time point). No occlusal interference was applied for control group. Bilateral masseter muscles of all the rats were acquired under general anesthesia. The samples of 5 rats in each group were fully homogenized with 0.4 mol/L perchlorate (10 mL/g). The homogenates were centrifuged, filtered and analyzed for ATP, ADP, IMP, phosphocreatine, creatine and lactate content by high performance liquid chromatography. The other samples in each group were mixed with homogenates containing 5 mmol/L sodium iodoacetate (10 mL/g), then homogenized and measured for pH value by pH meter in thermostatic water bathunder 37 degrees centigrade. RESULTS: Compared with control group, ATP content in bilateral masseter of the rats increased 3 d after occlusal interference [right side:(5.36±0.13) µmol/g,left side:(5.77±0.25) µmol/g] (P<0.05), and back to normal on 7, 10 and 14 d. There was an increase in IMP [right side:(0.21±0.03) µmol/g,left side:(0.19±0.03) µmol/g]and creatine content [right side:(24.76±2.94) µmol/g,left side:(27.75±2.23) µmol/g]in bilateral masseter of the rats 7 d after occlusal interference (P<0.05) and no difference was detected on 3, 10, and 14. Phosphocreatine content in bilateral masseter started to decline 7 d after occlusal interference and maintained the low level on 10 and 14 d [right side:(10.70±0.71) µmol/g, (11.57±0.52) µmol/g, (10.74±1.39) µmol/g, left side:(10.05±0.57) µmol/g, (10.75±1.12)µmol/g, (10.61±1.15) µmol/g](P<0.05). No change of ADP, lactate or pH level in bilateral muscles of the rats after occlusal interference was observed (P>0.05). CONCLUSION: Occlusal interference influences the content of energy metabolites in masticatory muscle of rats, which may be related to the pathological process of masticatory muscles induced by occlusal interference, such as muscle pain, dysfunction and altered fiber architecture.

Nuclear receptor ecdysone-induced protein 75 is required for larval-pupal metamorphosis in the Colorado potato beetle Leptinotarsa decemlineata (Say).

20-hydroxyecdysone (20E) and juvenile hormone (JH) are key regulators of insect development. In this study, three Leptinotarsa decemlineata Ecdysone-induced protein 75 (LdE75) cDNAs (LdE75A, B and C) were cloned from L. decemlineata. The three LdE75 isoforms were highly expressed just before or right after each moult. Within the fourth larval instar, they showed a small rise and a big peak 40 and 80 h after ecdysis. The expression peaks of the three LdE75s coincided with the peaks of circulating 20E levels. In vitro midgut culture and in vivo bioassay revealed that 20E and an ecdysteroid agonist halofenozide (Hal) enhanced LdE75 expression in the day 1 final larval instars. Conversely, a decrease in 20E by feeding a double-stranded RNA (dsRNA) against an ecdysteroidogenesis gene, Shade (LdSHD), repressed the expression of LdE75. Moreover, Hal upregulated the expression of the three LdE75s in LdSHD-silenced larvae. Thus, 20E pulses activate the transcription of LdE75s. Furthermore, ingesting dsE75-1 and dsE75-2 from a common fragment of the three isoforms successfully knocked down these LdE75s, and caused developmental arrest. Finally, knocking down LdE75s significantly repressed the transcription of three ecdysteroidogenesis genes, lowered the 20E titre and affected the expression of two 20E-response genes. Silencing LdE75s also induced the expression of a JH biosynthesis gene, increased JH titre and activated the transcription of a JH early-inducible gene. Thus, Ld E75s are required for larval-pupal metamorphosis and act mainly by modulating 20E and JH titres and mediating their signalling pathways.

Identification and differential expression of microRNAs in the ovaries of pigs (Sus scrofa) with high and low litter sizes.

Litter size affects profitability in the swine industry. Mammalian ovaries play important roles during reproduction, including ovulation and hormone secretion, which are tightly regulated by specific microRNAs (miRNAs). In this study, we investigated the effects of specific miRNAs on porcine litter size. We compared the ovarian miRNAs of Yorkshire pigs with high (YH) and low (YL) litter sizes using Solexa sequencing technology. We identified 327 and 320 miRNAs in the ovaries of YH and YL pigs respectively. A total of 297 miRNAs were co-expressed; 30 and 23 miRNAs respectively were specifically expressed in the two libraries. A total of 83 novel miRNAs were predicted; 37 specific miRNAs were obtained, of which 21 miRNAs were upregulated and 16 miRNAs were downregulated in YH compared with YL. Additionally, 19 628 and 19 250 target genes were predicted in the two libraries respectively. The results revealed that specific miRNAs (i.e., miR-224, miR-99a, let-7c, miR-181c, miR-214 and miR-21) may affect porcine litter size. The results of this study will help in gaining understanding of the role of miRNAs in porcine litter size regulation.

[Disturbed sleep, anxiety and stress are possible risk indicators for temporomandibular disorders with myofascialpain].

OBJECTIVE: To explore the relationship betweensleep/psychological distress and temporomandibular disorders (TMD) and to discuss the possibility of disturbed sleep and psychological distress as risk indicators in relation to TMD in a Chinese population. METHODS: The standardized and validated self-reported Chinese version questionnaires including Pittsburgh sleep quality index (PSQI) and depression, anxietyandstress scales-21 (DASS-21) were used to measure sleep quality and psychological distress. A total of 755 TMD patients (172 males and 583 females)with a mean age (29.99 ± 13.60) years were included in the study. The patients were divided into 7 diagnostic groups based on the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD), including Group I(exclusively myofascial pain), Group II(exclusively disc displacement), Group III (exclusively arthralgia or degenerative joint disease), Group IV (myofascial pain plus disc displacement), Group V (myofascial pain plus arthralgia or degenerative joint disease), Group VI (disc displacement plus arthralgia or degenerative joint disease) and Group VII (myofascial pain plus disc displacement plus arthralgia or degenerative joint disease). For statistical analysis, the patients were subsequently grouped into those with (181 patients) and without (574 patients) myofascial pain. Chi-square tests, independent-samples t test, partial correlation as well as stepwise Logistic regression analysis were used to analyze the data, using software SPSS 20.0 and P<0.05 was of significance. RESULTS: The prevalence of moderate to (extremely) severe disturbed sleep and psychological distress was significantly higher in the myofascial pain group (27.1%, 28.7%, 60.8% and 32.0%) than in the non-myofascial pain group (disc displacement and arthralgia or degenerative joint disease, 11.1%, 10.1%, 27.4% and 11.0%, P<0.05).The comorbidity of self-reported disturbed sleep and psychological distress was significantly higher in the myofascial pain patients than those without myofascial pain (P<0.05). Stepwise logistic regression analysis demonstrated that disturbed sleep (OR=1.74), more specifically, subjective sleep quality (OR=1.69) and sleep disturbance (OR=1.63) respectively, anxiety (OR=3.14) and stress (OR=2.15) were possible risk indicators for myofascial pain, and the results were still significant even after controlling for age, sex, educational level, disease duration, sleep quality, depression, anxiety and stress respectively (P<0.05). CONCLUSION: Disturbed sleep and psychological distress symptoms are common in TMD patients. Disturbed sleep, anxiety and stress are possible risk indicators for myofascial pain, compared with disc displacement and arthralgia or degenerative joint diseases.

[Disturbed sleep, anxiety and stress are possible risk indicators for temporomandibular disorders with myofascialpain].

OBJECTIVE: To explore the relationship betweensleep/psychological distress and temporomandibular disorders (TMD) and to discuss the possibility of disturbed sleep and psychological distress as risk indicators in relation to TMD in a Chinese population. METHODS: The standardized and validated self-reported Chinese version questionnaires including Pittsburgh sleep quality index (PSQI) and depression, anxietyandstress scales-21 (DASS-21) were used to measure sleep quality and psychological distress. A total of 755 TMD patients (172 males and 583 females)with a mean age (29.99 ± 13.60) years were included in the study. The patients were divided into 7 diagnostic groups based on the Research Diagnostic Criteria for Temporomandibular Disorders (RDC/TMD), including Group I(exclusively myofascial pain), Group II(exclusively disc displacement), Group III (exclusively arthralgia or degenerative joint disease), Group IV (myofascial pain plus disc displacement), Group V (myofascial pain plus arthralgia or degenerative joint disease), Group VI (disc displacement plus arthralgia or degenerative joint disease) and Group VII (myofascial pain plus disc displacement plus arthralgia or degenerative joint disease). For statistical analysis, the patients were subsequently grouped into those with (181 patients) and without (574 patients) myofascial pain. Chi-square tests, independent-samples t test, partial correlation as well as stepwise Logistic regression analysis were used to analyze the data, using software SPSS 20.0 and P<0.05 was of significance. RESULTS: The prevalence of moderate to (extremely) severe disturbed sleep and psychological distress was significantly higher in the myofascial pain group (27.1%, 28.7%, 60.8% and 32.0%) than in the non-myofascial pain group (disc displacement and arthralgia or degenerative joint disease, 11.1%, 10.1%, 27.4% and 11.0%, P<0.05).The comorbidity of self-reported disturbed sleep and psychological distress was significantly higher in the myofascial pain patients than those without myofascial pain (P<0.05). Stepwise logistic regression analysis demonstrated that disturbed sleep (OR=1.74), more specifically, subjective sleep quality (OR=1.69) and sleep disturbance (OR=1.63) respectively, anxiety (OR=3.14) and stress (OR=2.15) were possible risk indicators for myofascial pain, and the results were still significant even after controlling for age, sex, educational level, disease duration, sleep quality, depression, anxiety and stress respectively (P<0.05). CONCLUSION: Disturbed sleep and psychological distress symptoms are common in TMD patients. Disturbed sleep, anxiety and stress are possible risk indicators for myofascial pain, compared with disc displacement and arthralgia or degenerative joint diseases.