[Porphyromonas gingivalis outer membrane vesicles activate Toll-like receptor 2 to promote osteoclast differentiation by carrying lipopolysaccharide].

Objective: To investigate the effects of Porphyromonas gingivalis derived outer membrane vesicles (Pg OMV) on osteoclast differentiation of macrophages and its underlying mechanisms. Methods: The morphology and the size distribution of Pg OMV were analyzed by transmission electron microscopy and nanoparticle tracing analysis, respectively. The osteoclast precursors were treated with 1, 3 and 10 mg/L Pg OMV (1, 3 and 10 mg/L OMV treatment group) or phosphate buffer solution (PBS)(control group). The formation of osteoclasts was analyzed by tartrate-resistant acid phosphase (TRAP) staining and F-actin staining and real-time quantitative PCR (RT-qPCR) were used to detect the expression of Fos and matrix metallopeptidase 9 (MMP9). Polymyxin B (PMB) was used to block lipopolysaccharide (LPS) and then Pg OMV was used to treat osteoclast precursor (PMB-OMV treatment group), and OMV treatment group was used as control. TRAP and F-actin staining were used to observe the formation of osteoclasts and actin rings. The effect of Pg OMV on the expression of Toll-like receptor (TLR) 2 and TLR4 in preosteoclasts was detected by Western blotting. The osteoclast precursors were pretreated with 10, 50, 100 and 200 µmol/L C29, an inhibitor of TLR2, and then treated with Pg OMV(OMV+10, 50, 100 and 200 µmol/L C29 treatment group) and OMV treatment group without C29 pretreatment was control. TRAP and F-actin staining were used to observe the formation of osteoclasts and actin rings. The osteoclast precursor cells were treated with OMV (OMV treatment group) and OMV incubated with PMB (PMB-OMV treatment group) and the expression of TLR2 in osteoclast precursor was detected by Western blotting. Results: Pg OMV showed classical vesicular structures, and the average particle size of Pg OMV were 179.2 nm. A large number of actin rings were observed in the 3 and 10 mg/L OMV treatment groups. The percentages of TRAP-positive osteoclast area in 3 mg/L OMV treatment group [(22.6±2.1)%] and 10 mg/L OMV treatment group [(32.0±2.3)%] were significantly increased compared with control group [(4.9±0.5)%] (P<0.001). Compared with the control group (1.000±0.029), the mRNA relative expression of Fos in 3 mg/L OMV treatment group (1.491±0.114) and 10 mg/L OMV treatment group (1.726±0.254) was significantly increased (P=0.013, P=0.001). Compared with the control group (1.007±0.148), the mRNA relative expression of MMP9 in the group of 10 mg/L OMV (2.232±0.097) was significantly increased (P<0.001). Actin ring formation was less in PMB-OMV treatment groups than in OMV treatment groups. The proportion of TRAP-positive osteoclasts area [(14.8±3.8)%] in PMB-OMV treatment group was significantly lower than OMV treatment group [(31.5±6.7) %] (P=0.004). The relative expression of TLR2 in OMV treatment group (1.359±0.134) was significantly higher than that in the control group (1.000±0.000) (t=4.62, P=0.044). Compared with the OMV treatment group [(29.4±1.7)%], 50, 100 and 200 µmol/L C29 significantly decreased the formation of osteoclasts [(24.0±1.7)%, (18.5±2.1)%, (9.1±1.3) %] (P=0.026, P<0.001, P<0.001). TLR2 protein expression in PMB-OMV group (0.780±0.046) was significantly lower than that in OMV group (1.000±0.000)(t=8.32, P=0.001). Conclusions: Pg OMV can promote osteoclast differentiation by carrying LPS, TLR2 plays an important role in Pg OMV mediated osteoclast differentiation.

[Periodontal tissue regeneration: current therapeutic strategies and future directions in further research].

Chronic and progressive destruction/damage of the periodontal tissues resulted from periodontitis is the leading cause of tooth loss in adults. Traditional periodontal therapies such as scaling and root planning or flap surgery have demonstrated effective in controlling local inflammation and in suppressing/arresting the disease progression of periodontitis. However, those infection control measures cannot help to regenerate lost periodontal tissues to a statistically or clinically significant degree. Although some successes regarding the reduction of the intrabony defect and maintenance of the periodontal homeostasis have been achieved in periodontal regenerative procedures, comprising but not limited to guided tissue regeneration (GTR) or bone grafting technique, the restorative effectiveness of the architecture and function of the lost or injured tissues is far from our clinical expectation. The use of the concept, technique, and method of tissue engineering for periodontal regeneration is a hotspot and animal studies have shown interesting outcomes in terms of functional regeneration of lost/damaged support tissues in the periodontium, including alveolar bone, periodontal ligament, and cementum. However, numerous issues need to be addressed before those regenerative approaches can be responsibly transformed to novel clinical therapies. Recently, paradigm that induces homing of host stem cells to site of the periodontium and encourage the body's innate capability to repair is a new research field termed endogenous regeneration. Given that endogenous regenerative technique avoids ex-vivo cell culture and transplantation, it should be relatively easier to be used in the treatment of clinical patients. Due to the limited oral microenvironment and harsh periodontal local condition for tissue regeneration, as well as poor understanding of periodontal regenerative biology, there is still a long way ahead to explore new effective, practical, and economical therapies to save and protect natural tooth and for combating highly prevalent periodontal disease.

[Nonimpacted third molars and the periodontal homeostasis of their adjacent second molars].

Due to the limitations of eruption time and space, third molars (M3s) are often impacted and induce a variety of oral diseases, bringing adverse effects on the health of their adjacent second molars (M2s). For a long time, a large number of studies have focused on the harm of impacted M3s (I-M3s) to the health of their adjacent teeth, while less attention has been paid to nonimpacted M3s (N-M3s) that have already erupted. In recent years, however, a growing number of studies and evidences have shown that the existence of N-M3s is also an important risk factor for various diseases of their adjacent teeth, whose hazard has not been taken seriously by dentists and patients. Based on the latest results of both domestic and international researches as well as our group, this review summarizes and explains the effects of N-M3s on the periodontal homeostasis and periodontal health of adjacent M2s, so as to provide reference for clinical decision-making of N-M3s and the healthy maintenance of their adjacent teeth.

[Removal or retention of nonimpacted third molars].

Nonimpacted third molars (N-M3s) refer to the third molars (M3s) that completely erupt and reach the occlusal plane without rotation. Although the removal of unerupted M3s in young adults is generally approved by most dentists, the decision on removal or retention of N-M3s remains challenging as a result of unawareness of the potential harm of N-M3s. Recent studies suggested that N-M3s were associated with deteriorated effects to the health of adjacent teeth, including higher risks of caries and alveolar bone resorption. Therefore, regular monitoring and early clinical decision on N-M3s are essential to reduce the risks of adverse effects on adjacent teeth. This review is structure to give a comprehensive overview of the negative impact of N-M3s to the adjacent teeth, in order to provide rational guidelines for clinical decision-making of N-M3s.

[Long non-coding RNA LINC01133 regulates cementogenic differentiation of human periodontal ligament stem cells by modulating mitochondrial functions].

Objective: To investigate the effects of long non-coding RNA (lncRNA) LINC01133 on the cementogenic differentiation of human periodontal ligament stem cells (hPDLSC) and the underlying mechanism. Methods: A total of 12 teeth were harvested from 10 patients aged 17-30 years in the Department of Oral and Maxillofacial Surgery, School of Stomatology, The Fourth Military Medical University for impacted or orthodontic reasons from September 2021 to January 2022. The hPDLSCs were isolated from the teeth and transfected with small interfering RNA-LINC01133 (si-LINC01133) or small interfering RNA-negative control (si-NC). The si-LINC01133 was regarded as the experimental group, and the si-NC was regarded as the control one. The silencing efficiency of LINC01133 in the hPDLSCs was evaluated by real-time quantitative PCR (RT-qPCR). Western blotting was used to detect the protein expression levels of cementogenic differentiation-related factors including bone sialoprotein (BSP), cementum attachment protein (CAP), and cementum protein-1 (CEMP-1). Mitochondrial reactive oxygen species (mtROS) production was assessed using the MitoSox by flow cytometry. Mitochondrial membrane potential (MMP) was detected by JC-1 fluorescence staining. Mitochondrial respiratory chain complexes proteins including NADH dehydrogenase [ubiquinone] 1 beta subcomplex subunit 8 (NDUFB8), succinate dehydrogenase complex flavoprotein subunit A (SDHA), ubiquinol-cytochrome c reductase core protein 1 (UQCR1), cytochrome c oxidase subunit 4 isoform 1 (COXⅣ), and ATP synthase F1 subunit alpha (ATP5A) were evaluated by Western blotting. Results: The expression levels of LINC01133 could be suppressed by more than 60% with si-LINC01133 (control group: 1.000±0.000, experimental group: 0.385±0.128) (t=10.72, P<0.01). Suppression of LINC01133 in hPDLSCs decreased the levels of cementogenic differentiation-related proteins including BSP (control group: 1.000±0.000, experimental group: 0.664±0.179) (t=4.62, P<0.01) and CAP (control group: 1.000±0.000, experimental group: 0.736±0.229) (t=2.83, P<0.05). Suppression of LINC01133 in hPDLSCs increased the production of mtROS (control group: 1.000±0.000, experimental group: 1.458±0.185) (t=4.96, P<0.05) and the expression of NDUFB8 (control group: 1.000±0.000, experimental group: 1.683±0.397) (t=3.45, P<0.05), as well as decreased MMP levels (control group: 1.000±0.000, experimental group: 0.209±0.029) (t=53.99, P<0.01) and the expression of SDHA (control group: 1.000±0.000, experimental group: 0.428±0.228) (t=5.02, P<0.05). No significant changes in the UQCR1, COXⅣ, and ATP5A expression levels were found between the control group and exprimental group (P>0.05). Conclusions: LINC01133 regulates the cementogenic differentiation of hPDLSCs possibly via modulating the mitochondrial functions.

[Research progress in the association of periodontitis and gestational diabetes mellitus].

Periodontitis is the main cause of adult tooth loss, which seriously affects oral health and acts as a high-risk factor for varieties of systemic diseases. Gestational diabetes mellitus (GDM) is defined as glucose intolerance occurred or firstly identified during pregnancy. Prevalence of GDM is increasing over the past years worldwide. Besides adverse effects toward maternal and infant health in perinatal period, GDM also has long-term effects. Current studies have demonstrated that there is a bidirectional relationship between periodontitis and diabetes; however, the exact relationship between periodontitis and GDM remains elusive. In this paper, first reviewed the clinical association of periodontitis and GDM, and then discussed the underlying mechanisms of the two diseases, finally summarized the positive effect of periodontal therapy in controlling GDM. This paper will provide theoretical basis for the prevention diagnosis and therapy for the related diseases, promoting the maternal and infant health.

[Inflammatory factors in periodontitis patients and their effects toward the occurrence of gestational diabetes mellitus: a case-control study].

Objective: To explore the effects of periodontitis and inflammatory factors toward the occurrence of gestational diabetes mellitus (GDM). Methods: Pregnant women who came to the Department of Obstetrics, Northwest Women's and Children's Hospital for prenatal examinations during March to November of 2021 were invited to participate in this study. Participants with GDM who met the inclusion criteria (n=100) were assigned into the case group; while healthy participants (n=100) were assigned into the control group. Information of participants from the two groups were collected by questionnaires and periodontal statuses were clinically recorded in the meantime. Gingival crevicular fluid (GCF) and venous blood were also collected from participants of two groups to analyze the expression levels of inflammatory factors like C-reactive protein (CRP), tumor necrosis factor-α (TNF-α), interleukin (IL)-1ß, IL-6, IL-8, IL-10 and IL-33. Factors different between the two groups were included in the multivariate regression analysis model to determine the risk factors of GDM. Results: The age of participants was (33.4±5.1) years in case group and (30.5±4.5) years in control group respectively, which had statistical differences (t=4.33,P<0.001). Besides, the body mass index of participants from case group was also significantly higher than control group [(28.11±3.85) kg/m2 vs. (23.31±3.15) kg/m2, t=9.65, P<0.001]. Participants with GDM had more adverse periodontal clinical parameters. Prevalence of periodontitis in GDM group was 47.0% (47/100) compared with 29.0% (29/100) in control group (χ²=6.88, P=0.009). Multivariate regression analysis results indicated that periodontitis was a critical risk factor for the occurrence of GDM (OR=1.882, P<0.001). Besides, GCF IL-8, serum TNF-α, IL-8 and IL-10 were also risk factors of GDM due to their higher expressions. Among them, TNF-α in serum (OR=2.077) and IL-8 in serum (OR=2.060) had more significant impacts (P<0.001). Conclusions: This study demonstrated that periodontitis was associated with the occurrence of GDM. Up-regulation of serum pro-inflammatory mediators leaded by local periodontal inflammatory microenvironment might play a critical role in this pathological process.

[Effects of obstructive sleep apnea hypopnea syndrome on cardiac function in patients with chronic obstructive pulmonary disease].

Objective: To analyze the clinical characteristics of patients with chronic obstructive pulmonary disease (COPD) and COPD overlapping obstructive sleep apnea hypopnea syndrome (overlap syndrome), and to study the relationship between overlap syndrome and cardiovascular diseases. Methods: A total of 126 stable COPD patients admitted to the Respiratory Department of Peking University Third Hospital from September 2016 to October 2018 were included in this study, including 112 males and 14 females, ranging in age from 48 to 89 years, with a median of 67 years. With apnea hypopnea index (AHI) 5 times/h for the cutoff value, we classified the patients into a simple COPD group (31 cases) and an overlap syndrome group (95 cases), and compared the patients' demographic characteristics, respiratory symptoms, lung function, the incidence of cardiovascular events and the cardiac function with echocardiography (E/e'), left atrium diameter (LAD) and left ventricular ejection fraction (LVEF), by using independent-samples T test and chi-square test. Results: There were no statistically significant differences in demographic characteristics, respiratory symptoms, pulmonary function, cardiac function between COPD patients and overlap syndrome patients, but significant differences in blood oxygen level at night and left ventricular mass index(LVMI) between these groups (P=0.014,P<0.001,P<0.001,P<0.001, P=0.047, respectively) were observed. By comparing the severe sleep apnea hypopnea syndrome (AHI≥30) with sleep apnea hypopnea syndrome patients(AHI<30), there were statistically significant differences in echocardiographic indicators, among which there were statistically significant differences in E/e'(P=0.013), LAD(P=0.006), LVMI (P=0.051) and LVEF (P=0.030).There were also significant differences in the history of coronary heart disease and congestive heart failure between the two groups (P=0.025, P<0.001). After dividing the patients with overlap syndrome by mild, moderate and severe severity, E/e' and LAD were significantly correlated with severity (P=0.045, P=0.011). In terms of blood oxygen level at night, there was a significant correlation between average blood oxygen saturation at night and E/e', LAD, and LVMI (r=-0.195, P=0.033; r=-0.197, P=0.030; r=-0.195, P=0.044); moreover, there was also a significant correlation between the ratio of blood oxygen≤90% and LAD (r=0.209, P=0.021). In the multiple linear regression model, E/e' increased by 0.070 on average for each unit increase in AHI, and 0.084 on average for each unit increase in oxygen desaturation index (ODI). Conclusions: Patients with COPD overlapping severe sleep apnea hypopnea syndrome showed worse left diastolic function and higher risk of congestive heart failure and coronary heart disease compared with the patients with COPD alone. In addition, the degree of impairment of left heart diastolic function was associated with the severity of COPD overlapping sleep apnea hypopnea syndrome. The higher the AHI and the ODI became, the more severe the left heart diastolic restriction and structures changed.

Comparative proteomic analysis of different developmental stages of swamp buffalo testicular seminiferous tubules.

With ageing, many protein components change markedly during mammalian spermatogenesis. Most of these proteins have yet to be characterized and verified. Here, we have employed two-dimensional electrophoresis coupled to tandem mass spectrometry to explore the different proteins from pre-pubertal, pubertal and post-pubertal swamp buffalo testicular seminiferous tubules. The results showed that 25 protein spots were differentially expressed among developmental stages, and 13 of them were successfully identified by mass spectrometry. Of which four proteins were up-regulated and three proteins were down-regulated with age, and the remaining six proteins were fluctuated among developmental stages. Bioinformatics analysis indicates that these proteins were probably related to cellular developmental process (53.8%), cell differentiation (53.8%), spermatogenesis (15.4%), apoptotic process and cell death (30.8%). Expression profiles of calumenin (CALU) and galectin-1 (LGALS1) were further verified via Western blotting. In summary, the results help to develop an understanding of molecular mechanisms associated with buffalo spermatogenesis.

Leveraging Stem Cell Homing for Therapeutic Regeneration.

Resident stem cell pools in many tissues/organs are responsible not only for tissue maintenance during physiologic turnover but also for the process of wound repair following injury. With inspiration from stem cell trafficking within the body under physiologic and pathologic conditions, recent advances have been made toward inducing stem cell mobilization and directing patients' own cells to sites of interest for treating a broad spectrum of diseases. An evolving body of work corroborates that delivering guidance cues can mobilize stem cells from the bone marrow and drive these cells toward a specific region. In addition, the transplantation of cell-friendly biomaterials incorporating certain biomolecules has led to the regeneration of lost/damaged tissue without the need for delivering cellular materials manipulated ex vivo. Recently, cell homing has resulted in remarkable biological discoveries in the laboratory as well as great curative successes in preclinical scenarios. Here, we review the biological evidence underlying in vivo cell mobilization and homing with the aim of leveraging endogenous reparative cells for therapeutic applications. Considering both the promise and the obstacles of this approach, we discuss how matrix components of the in vivo milieu can be modified to promote the native regenerative process and inspire future tissue-engineering design.

Dynamic modelling of strategies for the control of acute haemorrhagic conjunctivitis outbreaks in schools in Changsha, China (2004-2015).

Outbreaks of acute haemorrhagic conjunctivitis (AHC) - a rapidly progressing and highly contagious infection - often occur in schools during summer and autumn. We used dynamic modelling to evaluate the efficacy of interventions to control AHC outbreaks in schools. A susceptible-infected-recovered (SIR) model was built to simulate AHC outbreaks in Chinese schools, with isolation or school closure added into the model. We used outbreak data from the period 2004-2015 in our models to estimate the effective reproduction number and assess the efficacy of interventions. The median effective reproduction number (uncontrolled) of AHC outbreaks was 7·00 (range 1·77-25·87). The median effective reproduction number (controlled) of AHC outbreaks was 0·16 (range 0·00-2·28). Intervention efficacy is affected by the timing of isolation; earlier isolation is associated with a lower morbidity peak and smaller total attack rate (TAR). School closures were not effective; TARs were almost 100% and did not change even when different school closure durations were adopted. Isolation and school closure as a combined intervention strategy was used to simulate outbreak control, but the efficacy was the same as isolation alone. An isolation programme could be an effective primary intervention during AHC outbreaks in schools. However, school closure is not recommended.

[Periodontal tissue engineering and regeneration].

The clinical management of periodontal disease is a global concern, and the regeneration of periodontal tissue defects due to periodontitis faces a huge challenge in the field of regenerative dentistry. Although conventional periodontal therapies focusing on in flammation control could stop or delay the progression of the disease, periodontal regeneration remains an elusive but laudable goal. Since late 1980s, concerted efforts have been made to accelerate and augment periodontal repair by using guided tissue regeneration (GTR), guided bone regeneration (GBR) and a wide range of other regenerative paradigms. Those advances have largely improved the clinical outcomes of periodontal therapies. In the past several years of 21st century, many progresses were made in the developments of stem cell therapy and tissue engineering, including remarkable biological discoveries in the laboratory as well as great curative successes in preclinical scenarios. The use of the principles, techniques and procedures of tissue engineering in periodontology showed great potential to regenerate new functional periodontal tissues such as alveolar bone, periodontal ligament, root cementum and finally and predictably the normal structure and functionality of the periodontium around a previously diseased tooth.

[Effect of Tripterygium wilfordii polycoride upon inflammation and TLR4/MyD88 signaling pathway in ulcerative colitis rats model].

OBJECTIVE: To observe the effect of Tripterygium wilfordii polycoride (TWP) on ulcerative colitis (UC), and its intervention effect on toll-like receptor 4 (TLR4)/myeloid differentiation factor 88 (MyD88) signaling pathway, thus to investigate its possible mechanism. METHODS: Trinitrobenzene sulfonic acid (TNBS)/ethanol enema method was used to set up the UC rat model. With random number table, 90 male Wistar rats were divided into normal control group, model group, TWP low, medium and high dose group (3, 6, 12 mg/kg, respectively) and azathioprine (AZA) group (6 mg/kg), with 15 rates in each group. Four days after enema, rates in each group were given corresponding drug lavage for 14 consecutive days. Disease activity index (DAI), colon gross morphological damage and histological grading of each group were observed. Using Western blot and reverse transcription (RT)-PCR method, the TLR4/MyD88 signaling pathway-related proteins in UC rat intestinal tissue were detected, namely TLR4, MyD88, tumor necrosis factor receptor related factor 6 (TRAF-6), nuclear factor kappa B (NF-κB), tumor necrosis factor alpha (TNF-α), and interleukin-1 beta (IL-1ß). RESULTS: The DAI, colon gross morphological damage, and histological grading of the model group were significantly higher than that of the normal control group (all P<0.01), indicating successful establishment of UC model. The DAI, colon gross morphological damage and histological grading of the TWP high dose group were lower than those of the model group (0.87±0.25 vs 1.60±0.76, 3.93±1.94 vs 5.40±2.21, 5.45±2.73 vs 13.27±3.50, P<0.05). Compared with the normal control group, the mRNA and protein expressions of TLR4, MyD88, TRAF-6, NF-κB, TNF-α, and IL-1ß in the model group rats were significantly increased (all P<0.01); which were significantly decreased in the TWP high dose group compared with model group rats (mRNA: 2.166±0.475 vs 5.647±0.275, 1.295±0.087 vs 3.774±0.418, 1.125±0.188 vs 2.535±0.320, 1.201±0.152 vs 2.082±0.077, 1.525±0.218 vs 3.094±0.022, 1.797±0.257 vs 17.152±0.145; protein: 0.252±0.010 vs 0.277±0.008, 0.172±0.002 vs 0.213±0.005, 0.233±0.006 vs 0.248±0.003, 0.099±0.003 vs 0.122±0.007, 0.238±0.002 vs 0.252±0.005, 0.235±0.003 vs 0.245±0.006, all P<0.05), also decreased in the AZA group (all P<0.01); and there were no significant differences between the TWP high dose group and the AZA group (all P>0.05). CONCLUSIONS: TWP can alleviate intestinal inflammation, promote healing of mucosa, showing a therapeutic effect for UC. One of its mechanisms may be through inhibiting the expression of TLR4, affecting the expression of TRAF-6, which is downstream to MyD66 signaling pathway, thus to suppress the activation of NF-κB and reduce the release of inflammatory factor such as TNF-α and IL-1ß.

[Clinical auxiliary diagnosis value of high frequency ultrasonographic measurements of the thickness of transverse carpal ligaments in carpal tunnel syndrome patients].

OBJECTIVE: To evaluate the meaning and value of high-frequency ultrasound in the diagnosis of carpal tunnel syndrome (CTS). METHODS: In this study, 48 patients (unilateral hand) with CTS were analyzed. The thickness of transverse carpal ligaments at the pisiform bone was measured using high-frequency ultrasound. Open carpal tunnel release procedure was performed in the 48 CTS patients, and the thickness of transverse carpal ligaments at the hamate hook bone measured using vernier caliper under direct vision. The accuracy of thickness of transverse carpal ligaments was evaluated using high-frequency ultrasound. high-frequency ultrasound measurement of thickness of transverse carpal ligaments at the hamate hook bone and pisiform bone, and determination of the diagnostic threshold measurement index using receiver operating characteristic (ROC) curve, sensitivity and specificity were performed and the correlation between the thickness of transverse carpal ligaments and nerve conduction study (NCS) analyzed. RESULTS: The thickness of transverse carpal ligaments in the CTS patients were (0.42±0.08) cm (high-frequency ultrasound) and (0.41±0.06) cm (operation) at hamate hook bone, and there was no significant difference between the two ways (t=0.672, P>0.05). The optimal cut-off value of the transverse carpal ligaments at hamate hook bone was 0.385 cm, the sensitivity 0.775, and the specificity 0.788. The optimal cut-off value of the transverse carpal ligaments at the pisiform bone was 0.315 cm, the sensitivity 0.950, and the specificity 1.000. The transverse carpal ligaments thickness and wrist-index finger sensory nerve conduction velocity (SCV), wrist-middle finger SCV showed a negative correlation. CONCLUSION: High frequency ultrasound measurements of thickness of transverse carpal ligaments is a valuable method for the diagnosis of CTS.

Efficacy evaluation of clonazepam for symptom remission in burning mouth syndrome: a meta-analysis.

OBJECTIVES: Clonazepam has been used in the treatment of burning mouth syndrome (BMS) for several decades. We conducted a meta-analysis to investigate the efficacy of clonazepam in the treatment of BMS. METHODS: We conducted a search of the PubMed, MEDLINE, EMBASE, Web of Science (TS), and the Cochrane Library databases for relevant studies that met our eligibility criteria (up to September 22, 2015). Statistical analyses were conducted using RevMan 5.2 and STATA 11.0 software. RESULTS: Three randomized controlled trials (RCTs) and two high-quality case-control studies involving 195 BMS patients were selected for this study. Our results show that clonazepam can reduce the oral pain sensation in patients with BMS (WMD: -3.72, 95% CI: -4.57, -2.86; P < 0.05; for all five studies). A positive therapeutic effect was demonstrated for both short-term (≤10 weeks) application (WMD: -1.44, 95% CI: -2.06, -0.82; P < 0.05) and long-term (>10 weeks) application (WMD: -4.50, 95% CI: -4.98, -4.03; P < 0.05). Both topical (WMD: -1.50, 95% CI: -2.14, -0.85; P < 0.05) and systemic (WMD: -3.81, 95% CI: -4.63, -2.98; P < 0.05) administration of clonazepam were confirmed to be effective. CONCLUSION: Clonazepam is effective in inducing symptom remission in patients with BMS.

Differences between high- and low-motility buffalo sperm identified by comparative proteomics.

This study was undertaken to investigate differences in protein expression between high- and low-motility sperm of swamp buffalo. The research used two-dimensional gel electrophoresis (2DE) coupled to matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry (MALDI-TOF/TOF-MS) to analyse the different proteins. The results showed 18 different expression protein spots between high- and low-motility buffalo sperm; eight of these proteins were up-regulated in low-motility sperm, five were down-regulated, one deleted and four proteins specifically expressed. Finally, four proteins were successfully identified by MS as belonging to three unique proteins; they are outer dense fibre of sperm tails protein 2 (ODF2), ATP synthase subunit alpha (ATP5A1) and succinyl-CoA synthetase subunit beta (SUCLG2). In summary, these results help to develop an understanding of the molecular mechanisms associated with low-motility sperm and provide clues for finding molecular markers associated with sperm motility.

Expression of ß-glucuronidase on the surface of bacteria enhances activation of glucuronide prodrugs.

Extracellular activation of hydrophilic glucuronide prodrugs by ß-glucuronidase (ßG) was examined to increase the therapeutic efficacy of bacteria-directed enzyme prodrug therapy (BDEPT). ßG was expressed on the surface of Escherichia coli by fusion to either the bacterial autotransporter protein Adhesin (membrane ßG (mßG)/AIDA) or the lipoprotein (lpp) outermembrane protein A (mßG/lpp). Both mßG/AIDA and mßG/lpp were expressed on the bacterial surface, but only mßG/AIDA displayed enzymatic activity. The rate of substrate hydrolysis by mßG/AIDA-BL21cells was 2.6-fold greater than by pßG-BL21 cells, which express periplasmic ßG. Human colon cancer HCT116 cells that were incubated with mßG/AIDA-BL21 bacteria were sensitive to a glucuronide prodrug (p-hydroxy aniline mustard ß-D-glucuronide, HAMG) with an half maximal inhibitory concentration (IC50) value of 226.53±45.4 µM, similar to the IC50 value of the active drug (p-hydroxy aniline mustard, pHAM; 70.6±6.75 µM), indicating that mßG/AIDA on BL21 bacteria could rapidly and efficiently convert HAMG to an active anticancer agent. These results suggest that surface display of functional ßG on bacteria can enhance the hydrolysis of glucuronide prodrugs and may increase the effectiveness of BDEPT.