[Establishment and application of new techniques for submandibular gland preservation].

The saliva secreted from submandibular gland (SMG) accounts for 60%-65%. It plays an important role in maintaining the human function of swallow, digestion, testing, speech, protection of oral mucosa, and prevention from dental carries. The SMG is frequently resected during the treatment for various kinds of oral and maxillofacial diseases, resulting in xerostomia and decreased quality of life. During the past 15 years, Research Center of Salivary Gland Diseases in Peking University School and Hospital of Stomatology conducted a series of studies on new techniques for preservation of SMG and achieved remarkable results. The clinicopathologic and imaging characteristics of IgG4-related sialadenitis (IgG4-RS) were clarified based on systematic studies. The results of studies on the pathogenesis of IgG4-RS showed that unbalance of inflammatory factors mediated the abnormality of secretion of SMG. IL-4 participates in occurring and development of glandular fibrosis of SMG. Regulation of tumor necrosis factor α (TNF-α) and cleaning of senescent cells might be taken as the targets for treatment of IgG4-RS. The combination of glucocorticoid and steroid-sparing agents showed effective results for treating IgG4-RS, clinical remission was achieved in all the patients, serum IgG4 levels decreased, and salivary gland secretion significantly increased. Sialoendoscopy-assisted sialolithectomy was applied in the treatment of about 1 000 cases with submandibular hilar calculi with a success rate of more than 90%. Transfer of SMG was used for prevention from radiation-induced xerostomia in the patients with head and neck carcinoma. SMG was transferred to the submental region before radiotherapy and was kept away from the ra-diation field. The results of prospective clinical controlled study showed this technique could effectively preserve the function of SMG and prevent from xerostomia. Based on the micro-anatomical study on the blood vessels and ducts of SMG, partial sialoadenectomy was applied for treatment of benign tumors in the SMG. A clinical controlled study confirmed its safety for control of the tumors and its advantage of preservation of SMG function. The studies on the involvement of SMG in oral squamous cell carcinoma (OSCC) provided the anatomical and histopathological basis for preservation of SMG during neck dissection for early cases with OSCC. A innovated surgical modality "four preservations including SMG" was used during the neck dissection for the early cases with OSCC. A prospective randomized clinical controlled study confirmed its safety, feasibility, effectiveness for control of the carcinoma, and advantages of preservation of SMG function.

[Role and mechanism of muscarinic acetylcholine receptor in the regulation of submandibular gland secretion].

Muscarinic acetylcholine receptors (mAChRs), including M1-M5 subtypes, are classic receptors in regulating water, ion, and solute transport in salivary gland. Our work focuses on the studies on the expression pattern and function of mAChR in the submandibular gland (SMG), and the underlying mechanism involved in the mAChR-regulated secretion, together with the effect of parasympathectomy on the salivary secretion. Microvascular autotransplantation of SMG into the temporal fossa provides a continuous and endogenous source of fluids, and is currently an effective method for treating severe keratoconjunctivitis sicca. By using RT-PCR, Western blotting, and immunofluorescence, our data demonstrated that the expression of M1 and M3 subtypes were decreased in latent period in rabbit SMG autotransplantation model, whereas carbachol stimulation promoted the salivary secretion, as well as M1 and M3 expressions. By contrast, mAChRs were hypersensitive in epiphora SMGs, whereas atropine gel and botulinum toxin A application significantly inhibited the hypersecretion in both animal models and patients. Furthermore, the possible intracellular signal molecules involved in the mAChR-modulated salivary secretion were explored. Activation of mAChR upregulated the expression of aquaporin 5 (AQP5), the main transporter that mediated water secretion through transcellular pathway, and led to AQP5 trafficking from lipid rafts to non-lipid microdomain. Extracellular signal-regulated kinase 1/2 (ERK1/2) was involved in the mAChR-regulated AQP5 content. mAChR activation also modulated the expression, distribution, and function of tight junction proteins, and increased paracellular permeability. ERK1/2/ß-arrestin2/clathrin/ubiquitin signaling pathway was responsible for the mAChR-regulated downregulation of tight junction molecule claudin-4. Cytoskeleton filamentous actin (F-actin) was also involved in the distribution and barrier function of epithelial tight junctions. Besides, endothelial tight junctions were opened by mAChR agonist-evoked salivation in the mice. Furthermore, parasympathetic denervation increased resting salivary secretion in the long terminrats and minipigs. Taken together, our work demonstrated that mAChR regulated saliva secretion via transcellular and paracellular pathways in SMG epithelium as well as tight junction opening in SMG endothelium. Modulation of mAChR might be a promising strategy to ameliorate SMG dysfunction.

Expression of interleukin-17 in primary Sjögren's syndrome and the correlation with disease severity: A systematic review and meta-analysis.

The aberrant expression of interleukin-17 (IL-17) has been reported in primary Sjögren's syndrome (pSS). Abnormalities in IL-17 can promote the production of pro-inflammatory cytokines and aggravate autoimmune disorders. The aim of this study was to investigate alterations of IL-17 in patients with pSS and explore the correlation between IL-17 and disease severity. Eight databases were searched for original studies reporting the expression of IL-17 in patients with pSS and controls. Eligible reports were included in the pooled analysis, and subgroup evaluations were performed according to different types of controls and IL-17 measurement methods. Newcastle-Ottawa Scale criteria were used to assess the risk of bias of the included studies. In total, 45 articles are included in the meta-analysis. The expression of IL-17 is significantly increased in patients with pSS compared to controls. Furthermore, patients with pSS without immunosuppressive treatment show markedly higher IL-17 levels. In addition, patients with pSS with positive rheumatoid factors tend to express a higher level of IL-17 than patients with negative rheumatoid factors. Negative correlations between IL-17 levels and ocular parameters are also found in patients with pSS. The results are similar after adjustment by "trim and fill" methods. In conclusion, the expression of IL-17 is obviously increased in patients with pSS, especially among those without immunosuppressive treatment. In addition, IL-17 level correlates with the disease severity of pSS. These findings demonstrate the significance of IL-17 overexpression in patients with pSS and may provide insights for the development of therapeutic interventions targeting IL-17 for pSS.

Indoor particle dynamics in a school office: determination of particle concentrations, deposition rates and penetration factors under naturally ventilated conditions.

Recently, the problem of indoor particulate matter pollution has received much attention. An increasing number of epidemiological studies show that the concentration of atmospheric particulate matter has a significant effect on human health, even at very low concentrations. Most of these investigations have relied upon outdoor particle concentrations as surrogates of human exposures. However, considering that the concentration distribution of the indoor particulate matter is largely dependent on the extent to which these particles penetrate the building and on the degree of suspension in the indoor air, human exposures to particles of outdoor origin may not be equal to outdoor particle concentration levels. Therefore, it is critical to understand the relationship between the particle concentrations found outdoors and those found in indoor micro-environments. In this study, experiments were conducted using a naturally ventilated office located in Qingdao, China. The indoor and outdoor particle concentrations were measured at the same time using an optical counter with four size ranges. The particle size distribution ranged from 0.3 to 2.5 µm, and the experimental period was from April to September, 2016. Based on the experimental data, the dynamic and mass balance model based on time was used to estimate the penetration rate and deposition rate at air exchange rates of 0.03-0.25 h-1. The values of the penetration rate and deposition velocity of indoor particles were determined to range from 0.45 to 0.82 h-1 and 1.71 to 2.82 m/h, respectively. In addition, the particulate pollution exposure in the indoor environment was analyzed to estimate the exposure hazard from indoor particulate matter pollution, which is important for human exposure to particles and associated health effects. The conclusions from this study can serve to provide a better understanding the dynamics and behaviors of airborne particle entering into buildings. And they will also highlight effective methods to reduce exposure to particles in office buildings.

[A 20-year study on microvascular autologous transplantation of submandibular gland for treatment of severe dry eye].

Severe dry eye is a refractory ophthalmologic disease. Our multidisciplinary research group treated severe dry eye by microvascular autologous transplantation of submandibular gland (SMG) during the past 20 years. The SMG, with its blood vessels and Wharton's duct, was harvested from the submandibular triangle and transferred to the temporal area. The blood vessels in the SMG were anastomosed with the temporal blood vessels using a microsurgical technique. Then, the distal end of Wharton's duct was sutured to form an opening in the upper lateral conjunctival fold. The tear was replaced by the secretion of the transplanted SMG to lubricate the ocular surface. In our study, the surgical techniques of blood vessel management were continuously modified to increase the survival rate of the transplanted SMG. A novel surgical modality of partial transplantation of SMG was established to prevent postoperative epiphora. A clinical study with the largest case number in the world was conducted and the effectiveness of transplantation of SMG for severe dry eye was fully confirmed. In order to resolve two main clinical problems including ductal obstruction resulted from low secretion rate during the latent period, and epiphora due to over secretion of the transplanted SMG in the later term of transplantation, the regulation of the secretion mechanism of the normal and transplanted SMG were investigated. New opinions on mechanisms of saliva secretion were provided. Based on the priniciple of translational medicine, the results of related basic research were applied in the clinic. The clinical guidelines for secretion regulation of transplanted SMG were established. A concept of chronic obstructive sialadenitis of transplanted SMG was provided and its diagnostic criteria, diagnostic technique of sialography, and therapeutic regimen were established. As a result, the surgical success rate was obviously elevated, the surgical complications were decreased, and life quality of the patients was greatly improved.

Biomarkers predict enhanced clinical outcomes with afatinib versus methotrexate in patients with second-line recurrent and/or metastatic head and neck cancer.

BACKGROUND: In the phase III LUX-Head & Neck 1 (LUX-H&N1) trial, second-line afatinib significantly improved progression-free survival (PFS) versus methotrexate in patients with recurrent/metastatic head and neck squamous cell carcinoma (R/M HNSCC). Here, we evaluated association of prespecified biomarkers with efficacy outcomes in LUX-H&N1. PATIENTS AND METHODS: Randomized patients with R/M HNSCC and progression following ≥2 cycles of platinum therapy received afatinib (40 mg/day) or methotrexate (40 mg/m2/week). Tumor/serum samples were collected at study entry for patients who volunteered for inclusion in biomarker analyses. Tumor biomarkers, including p16 (prespecified subgroup; all tumor subsites), EGFR, HER2, HER3, c-MET and PTEN, were assessed using tissue microarray cores and slides; serum protein was evaluated using the VeriStrat® test. Biomarkers were correlated with efficacy outcomes. RESULTS: Of 483 randomized patients, 326 (67%) were included in the biomarker analyses; baseline characteristics were consistent with the overall study population. Median PFS favored afatinib over methotrexate in patients with p16-negative [2.7 versus 1.6 months; HR 0.70 (95% CI 0.50-0.97)], EGFR-amplified [2.8 versus 1.5 months; HR 0.53 (0.33-0.85)], HER3-low [2.8 versus 1.8 months; HR 0.57 (0.37-0.88)], and PTEN-high [1.6 versus 1.4 months; HR 0.55 (0.29-1.05)] tumors. Afatinib also improved PFS in combined subsets of patients with p16-negative and EGFR-amplified tumors [2.7 versus 1.5 months; HR 0.47 (0.28-0.80)], and patients with p16-negative tumors who were EGFR therapy-naïve [4.0 versus 2.4 months; HR 0.55 (0.31-0.98)]. PFS was improved in afatinib-treated patients who were VeriStrat 'Good' versus 'Poor' [2.7 versus 1.5 months; HR 0.71 (0.49-0.94)], but no treatment interaction was observed. Afatinib improved tumor response versus methotrexate in all subsets analyzed except for those with p16-positive disease (n = 35). CONCLUSIONS: Subgroups of HNSCC patients who may achieve increased benefit from afatinib were identified based on prespecified tumor biomarkers (p16-negative, EGFR-amplified, HER3-low, PTEN-high). Future studies are warranted to validate these findings. CLINICAL TRIAL REGISTRATION: NCT01345682.

Afatinib versus methotrexate in older patients with second-line recurrent and/or metastatic head and neck squamous cell carcinoma: subgroup analysis of the LUX-Head & Neck 1 trial.

BACKGROUND: In the phase III LUX-Head & Neck 1 (LHN1) trial, afatinib significantly improved progression-free survival (PFS) versus methotrexate in recurrent and/or metastatic (R/M) head and neck squamous cell carcinoma (HNSCC) patients progressing on/after platinum-based therapy. This report evaluates afatinib efficacy and safety in prespecified subgroups of patients aged ≥65 and <65 years. PATIENTS AND METHODS: Patients were randomized (2:1) to 40 mg/day oral afatinib or 40 mg/m(2)/week intravenous methotrexate. PFS was the primary end point; overall survival (OS) was the key secondary end point. Other end points included: objective response rate (ORR), patient-reported outcomes, tumor shrinkage, and safety. Disease control rate (DCR) was also assessed. RESULTS: Of 483 randomized patients, 27% (83 afatinib; 45 methotrexate) were aged ≥65 years (older) and 73% (239 afatinib; 116 methotrexate) <65 years (younger) at study entry. Similar PFS benefit with afatinib versus methotrexate was observed in older {median 2.8 versus 2.3 months, hazard ratio (HR) = 0.68 [95% confidence interval (CI) 0.45-1.03], P = 0.061} and younger patients [2.6 versus 1.6 months, HR = 0.79 (0.62-1.01), P = 0.052]. In older and younger patients, the median OS with afatinib versus methotrexate was 7.3 versus 6.4 months [HR = 0.84 (0.54-1.31)] and 6.7 versus 6.2 months [HR = 0.98 (0.76-1.28)]. ORRs with afatinib versus methotrexate were 10.8% versus 6.7% and 10.0% versus 5.2%; DCRs were 53.0% versus 37.8% and 47.7% versus 38.8% in older and younger patients, respectively. In both subgroups, the most frequent treatment-related adverse events were rash/acne (73%-77%) and diarrhea (70%-80%) with afatinib, and stomatitis (43%) and fatigue (31%-34%) with methotrexate. Fewer treatment-related discontinuations were observed with afatinib (each subgroup 7% versus 16%). A trend toward improved time to deterioration of global health status, pain, and swallowing with afatinib was observed in both subgroups. CONCLUSIONS: Advancing age (≥65 years) did not adversely affect clinical outcomes or safety with afatinib versus methotrexate in second-line R/M HNSCC patients. CLINICAL TRIAL REGISTRATION: NCT01345682 (ClinicalTrials.gov).

[Application of diffusion-weighted intravoxel incoherent motion imaging in diagnosis of renal cell carcinoma subtypes].

OBJECTIVE: To evaluate the value of parameters derived from intravoxel incoherent motion diffusion-weighted magnetic resonance imaging in differentiating histopathological subtypes of renal cell carcinoma (RCC). METHODS: Between May 2014 and December 2015, a total of 69 patients who were surgically and pathologically diagnosed as renal cell carcinoma were recruited for the study. We examined 61 clear cell RCC (ccRCC), and 8 non-clear cell carcinoma (non-ccRCC, including 7 chromophobe RCC and 1 papillary RCC). All the ccRCC were divided into well differentiated group (n=46), moderately differentiated group (n=8), and poorly differentiated group (n=7). In addition to routine renal magnetic resonance imaging examination performed on a 3.0-Tesla MR system, all patients were imaged with axial intravoxel incoherent motion diffusion-weighted imaging. Using biexponential model, we calculated the diffusion coefficient (D), pseudodiffusion coefficient (D(*)), and perfusion fraction (f). RESULTS: The D and f values of the ccRCC were higher (each P<0.05) than that for non-ccRCC [D (1.29±0.30)×10(-3)mm(2)/s, D(*) (42.92±20.21)×10(-3)mm(2)/s, and f (35.71±6.61)% versus D (0.78±0.23)×10(-3)mm(2)/s, D(*) (32.60±11.33)×10(-3)mm(2)/s, and f (21.52±8.44)% ]. In the well differentiated group of ccRCC, we found D of (1.36±0.29)×10(-3)mm(2)/s, D(*) (38.39±18.51)×10(-3)mm(2)/s, and f (36.40±6.96)%. The D, D(*,) f values of moderately differentiated lesions were (1.10±0.24)×10(-3)mm(2)/s, (59.90±20.23)×10(-3) mm(2)/s, and (32.88±4.02)%, respectively, those of the poorly differentiated group were (1.03±0.16)×10(-3)mm(2)/s, (53.28±18.74)×10(-3)mm(2)/s, and (34.42±6.21)%. The well differentiated group of ccRCC showed a higher D value than the moderately differentiated and poorly differentiated groups (each P<0.05). D(*) values were significantly lower for the well differentiated group than for the moderately differentiated group (P<0.05). The sensitivity and specificity of D values were 90.2% and 87.5% when focusing on the differentiation of ccRCC. For the diagnosis of ccRCC, the sensitivity and specificity of f values were 98.4% and 75.0%, respectively. CONCLUSIONS: IVIM-DWI can provide certain reliable value in evaluating pathological subtype and differentiation degree of renal cell carcinomas. D and f values are useful to distinguish ccRCC from non-ccRCC. D value is also promising for estimating the differentiation degree of ccRCC, and to indicate the biological behavior of RCC.

[Genetic characterization of varicella zoster virus in Jilin province in 2014, China].

OBJECTIVE: To clarify the genotype of wild-type strains of varicella zoster virus (VZV) in Jilin province in 2014, and to discriminate between v-Oka vaccine strains and wild-type strains. METHODS: Vesicle fluid and throat swab samples were collected from 13 individuals with suspected VZV in Jilin province from January to December 2014. Viral DNA was extracted, the fragments of 15 open reading fragments (ORFs) were amplified by polymerase chain reaction (PCR), and viral genotypes were determined by single nucleotide polymorphisms (SNP). PCR restriction fragment length polymorphism (RFLP) was used to distinguish between wild-type strains and v-Oka vaccine strains. The results were analyzed with MEGA5 software, using the VZV reference strain sequences from GenBank. RESULTS: The 13 suspected samples included 5 males and 8 females, aged 11-27 years (mean: (16.69±5.48) years). Sampling was performed on days 0 to 3 of suspected infection. VZV strains were detected in 8 samples, all belonging to Clade 2. There was a synonymous mutation (T>C) in SNP18082 compared with the v-Oka vaccine strain. Analysis of PCR-RFLPs showed that all 8 positive samples were wild-type strains (PstⅠ(+)BglⅠ(+)SmaⅠ(-)). CONCLUSIONS: The study revealed that the VZV strains circulating in Jilin province in 2014 were wild-type strains belonging to Clade 2.

In vitro and in vivo anticancer effects of mevalonate pathway modulation on human cancer cells.

BACKGROUND: The increasing usage of statins (the 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitors) has revealed a number of unexpected beneficial effects, including a reduction in cancer risk. METHODS: We investigated the direct anticancer effects of different statins approved for clinical use on human breast and brain cancer cells. We also explored the effects of statins on cancer cells using in silico simulations. RESULTS: In vitro studies showed that cerivastatin, pitavastatin, and fluvastatin were the most potent anti-proliferative, autophagy inducing agents in human cancer cells including stem cell-like primary glioblastoma cell lines. Consistently, pitavastatin was more effective than fluvastatin in inhibiting U87 tumour growth in vivo. Intraperitoneal injection was much better than oral administration in delaying glioblastoma growth. Following statin treatment, tumour cells were rescued by adding mevalonate and geranylgeranyl pyrophosphate. Knockdown of geranylgeranyl pyrophosphate synthetase-1 also induced strong cell autophagy and cell death in vitro and reduced U87 tumour growth in vivo. These data demonstrate that statins main effect is via targeting the mevalonate synthesis pathway in tumour cells. CONCLUSIONS: Our study demonstrates the potent anticancer effects of statins. These safe and well-tolerated drugs need to be further investigated as cancer chemotherapeutics in comprehensive clinical studies.

Decreased submandibular adiponectin is involved in the progression of autoimmune sialoadenitis in non-obese diabetic mice.

OBJECTIVE: To investigate a possible role of adiponectin in the pathogenesis of autoimmune sialoadenitis in non-obese diabetic (NOD) mouse model of Sjögren's syndrome. MATERIALS AND METHODS: Expression of adiponectin and its receptors (AdipoR1/2) was detected by PCR, immunoblotting, or immunofluorescence. The level of adiponectin was quantified by ELISA. Adiponectin-related signaling molecules and pro-inflammatory cytokines were examined by PCR or immunoblotting. Apoptosis was evaluated by TUNEL staining, flow cytometry, and caspase 3 activation. RESULTS: Adiponectin and AdipoR1/2 mRNA and protein were expressed in submandibular glands. Adiponectin immunostaining was widely diffused in the cytoplasm of acinar and ductal cells. AdipoR1 was mainly distributed in acinar cytoplasm, while AdipoR2 was predominantly located at acinar cell membrane. Submandibular adiponectin levels were reduced during the progression of autoimmune sialoadenitis in 7-, 14-, and 21-week-old NOD mice, while AdipoR1/2 levels were unchanged. The levels of phosphorylated adenosine monophosphate-activated protein kinase, extracellular signal-regulated kinase 1/2, and p38 mitogen-activated protein kinase were decreased, while interferon (IFN)-γ and glandular apoptosis were temporally increased at all time points. Moreover, exogenous adiponectin supplement inhibited, whereas neutralizing endogenous adiponectin by its antibody promoted IFN-γ-induced apoptosis and caspase 3 activation in cultured submandibular acinar cells. CONCLUSIONS: Adiponectin plays a protective role on submandibular cells. Decreased adiponectin might promote glandular destruction in autoimmune sialoadenitis.

[Pristimerin enhances cisplatin-induced apoptosis in nasopharyngeal carcinoma cells via ROS-mediated deactivation of the PI3K/AKT signaling pathway].

OBJECTIVE: To explore the effect of pristimerin combined with cisplatin on proliferation and apoptosis of nasopharyngeal carcinoma cells. METHODS: CCK-8 assay was used to examine the survival rate of HNE-1 and CNE-2Z cells following treatment for 24 h with different concentrations of pristimerin, cisplatin or their combination. The changes in colony formation ability, apoptosis, and intracellular reactive oxygen species (ROS) levels of the treated cells were analyzed using colony formation assay and flow cytometry. Western blotting was performed to detect the changes in protein expressions in the cells. The effects of pre-treatment with NAC on proliferation, apoptosis, and PI3K/AKT signaling pathway were observed in pristimerin- and/or cisplatin-treated cells. RESULTS: Both pristimerin and cisplatin significantly lowered the survival rate of HNE-1 and CNE-2Z cells (P < 0.05). Compared with pristimerin or cisplatin alone, their combination more strongly inhibited survival and colony formation ability of the cells, increased cell apoptosis rate and intracellular ROS levels, upregulated the protein expressions of Bax, cleaved caspase-3, and cleaved PARP, and downregulated the protein expressions of Bcl-2, Mcl-1, PARP and p-PI3K and p-AKT (P < 0.05). NAC pretreatment significantly attenuated proliferation inhibition and apoptosis-promoting effects of pristimerin combined with cisplatin, and partially restored the downregulated protein expressions of p-PI3K and p-AKT in HNE-1 and CNE-2Z cells with the combined treatment (P < 0.05). CONCLUSION: Pristimerin can enhance cisplatin-induced proliferation inhibition and apoptosis in nasopharyngeal carcinoma cells, the mechanism of which may involve ROS-mediated deactivation of the PI3K/AKT signaling pathway.

A novel Slide-seq based image processing software to identify gene expression at the single cell level.

Analysis of gene expression at the single-cell level could help predict the effectiveness of therapies in the field of chronic inflammatory diseases such as arthritis. Here, we demonstrate an adopted approach for processing images from the Slide-seq method. Using a puck, which consists of about 50,000 DNA barcode beads, an RNA sequence of a cell is to be read. The pucks are repeatedly brought into contact with liquids and then recorded with a conventional epifluorescence microscope. The image analysis initially consists of stitching the partial images of a sequence recording, registering images from different sequences, and finally reading out the bases. The new method enables the use of an inexpensive epifluorescence microscope instead of a confocal microscope.

Claudin-10 Decrease in the Submandibular Gland Contributes to Xerostomia.

Tight junction proteins play a crucial role in paracellular transport in salivary gland epithelia. It is clear that severe xerostomia in patients with HELIX syndrome is caused by mutations in the claudin-10 gene. However, little is known about the expression pattern and role of claudin-10 in saliva secretion in physical and disease conditions. In the present study, we found that only claudin-10b transcript was expressed in human and mouse submandibular gland (SMG) tissues, and claudin-10 protein was dominantly distributed at the apicolateral membranes of acini in human, rat, and mouse SMGs. Overexpression of claudin-10 significantly reduced transepithelial electrical resistance and increased paracellular transport of dextran and Na+ in SMG-C6 cells. In C57BL/6 mice, pilocarpine stimulation promoted secretion and cation concentration in saliva in a dose-dependent increase. Assembly of claudin-10 to the most apicolateral portions in acini of SMGs was observed in the lower pilocarpine (1 mg/kg)-treated group, and this phenomenon was much obvious in the higher pilocarpine (10 mg/kg)-treated group. Furthermore, 7-, 14-, and 21-wk-old nonobese diabetic (NOD) and BALB/c mice were used to mimic the progression of hyposalivation in Sjögren syndrome. Intensity of claudin-10 protein was obviously lower in SMGs of 14- and 21-wk-old NOD mice compared with that of age-matched BALB/c mice. In the cultured mouse SMG tissues, interferon-γ (IFN-γ) downregulated claudin-10 expression. In claudin-10-overexpressed SMG-C6 cells, paracellular permeability was decreased. Furthermore, IFN-γ stimulation increased p-STAT1 level, whereas pretreatment with JAK/STAT1 antagonist significantly alleviated the IFN-γ-induced claudin-10 downregulation. These results indicate that claudin-10 functions as a pore-forming component in acinar epithelia of SMGs, assembly of claudin-10 is required for saliva secretion, and downregulation of claudin-10 induces hyposecretion. These findings may provide new clues to novel therapeutic targets on hyposalivation.

Regulation of circadian locomotor rhythm by neuropeptide Y-like system in Drosophila melanogaster.

Circadian rhythms in behaviour and physiology exist widely in animals, plants, fungi and cyanobacteria. Although much work has been carried out to characterize the endogenous clock circuit, the output signals coupling the circadian pacemaker to behaviour and physiology remain elusive. Here, we show that neuropeptide F (NPF), a homologue of mammalian neuropeptide Y, and its G protein-coupled receptor NPFR1 regulate the locomotor rhythm in Drosophila melanogaster. Flies with loss of function in NPF or NPFR1 were unable to ramp up their activity before lights off under light : dark (LD) cycles, and oscillations in npf/NPF and npfr1/NPFR1 were found to correlate temporally with the locomotor rhythm. Furthermore, NPF is expressed in clock neurones including dorsolateral neurones (LNd s) and ventrolateral neurones (LNv s), whereas NPFR1 is expressed in dorsal neurone 1 (DN1) and LNd s. These results show that NPF signalling is involved in the circadian locomotor rhythm in LD cycles.

Neuropsychiatric symptoms, seizure severity, employment, and quality of life of Jordanians with epilepsy.

RATIONALE: Depression and anxiety are more strongly associated with quality of life (QOL) than seizure frequency in several populations with epilepsy. However, QOL is culturally determined and may be influenced by cultural values and norms as well as local policies and resources. The goal of this study is to investigate the impact of neuropsychiatric symptoms and seizure severity on QOL and employment in people with epilepsy living in Jordan. METHODS: Seizure severity and complications, antiepileptic drug side effects, social stigma, neuropsychiatric symptoms, and mental health (MH-SF36) and physical health (PH-SF36) domains of QOL were assessed in 45 adult patients with epilepsy in a university neurology clinic. Multivariate regression analysis was used to evaluate the relationship between these variables and the quality of life of Jordanians with epilepsy. RESULTS: Neuropsychiatric symptoms, seizure frequency, and history of injury due to seizure were associated with the MH-SF36. However, earlier age of seizure onset, longer duration of epilepsy, unemployment, and history of chronic disease was associated with lower PH-SF36 scores. Furthermore, there were no differences in QOL, neuropsychiatric symptoms, and seizure frequency in Jordanians who were employed versus unemployed in this study. CONCLUSIONS: Neuropsychiatric symptoms were significantly associated with mental health-related QOL measures, but not with physically-related QOL measures, in Jordanians with epilepsy. For studies across populations, it is critical to separate mental health from physical health QOL measures. Furthermore, regional differences in culture and policy may more strongly influence employment status than individuals' experiences of epilepsy, neuropsychiatric symptoms, or QOL in some populations.

A high-throughput, high-quality plant genomic DNA extraction protocol.

The isolation of high-quality genomic DNA (gDNA) is a crucial technique in plant molecular biology. The quality of gDNA determines the reliability of real-time polymerase chain reaction (PCR) analysis. In this paper, we reported a high-quality gDNA extraction protocol optimized for real-time PCR in a variety of plant species. Performed in a 96-well block, our protocol provides high throughput. Without the need for phenol-chloroform and liquid nitrogen or dry ice, our protocol is safer and more cost-efficient than traditional DNA extraction methods. The method takes 10 mg leaf tissue to yield 5-10 µg high-quality gDNA. Spectral measurement and electrophoresis were used to demonstrate gDNA purity. The extracted DNA was qualified in a restriction enzyme digestion assay and conventional PCR. The real-time PCR amplification was sufficiently sensitive to detect gDNA at very low concentrations (3 pg/µL). The standard curve of gDNA dilutions from our phenol-chloroform-free protocol showed better linearity (R(2) = 0.9967) than the phenol-chloroform protocol (R(2) = 0.9876). The results indicate that the gDNA was of high quality and fit for real-time PCR. This safe, high-throughput plant gDNA extraction protocol could be used to isolate high-quality gDNA for real-time PCR and other downstream molecular applications.