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1.  Enhanced oral absorption of 20(S)-protopanaxadiol by self-assembled liquid crystalline nanoparticles containing piperine: in vitro and in vivo studies 
Background
20(S)-protopanaxadiol (PPD), similar to several other anticancer agents, has low oral absorption and is extensively metabolized. These factors limit the use of PPD for treatment of human diseases.
Methods
In this study, we used cubic nanoparticles containing piperine to improve the oral bioavailability of PPD and to enhance its absorption and inhibit its metabolism. Cubic nanoparticles loaded with PPD and piperine were prepared by fragmentation of glyceryl monoolein (GMO)/poloxamer 407 bulk cubic gel and verified using transmission electron microscopy and differential scanning calorimetry. We evaluated the in vitro release of PPD from these nanoparticles and its absorption across the Caco-2 cell monolayer model, and subsequently, we examined the bioavailability and metabolism of PPD and its nanoparticles in vivo.
Results
The in vitro release of PPD from these nanoparticles was less than 5% at 12 hours. PPD-cubosome and PPD-cubosome loaded with piperine (molar ratio PPD/piperine, 1:3) increased the apical to basolateral permeability values of PPD across the Caco-2 cell monolayer from 53% to 64%, respectively. In addition, the results of a pharmacokinetic study in rats showed that the relative bioavailabilities of PPD-cubosome [area under concentration–time curve (AUC)0–∞] and PPD-cubosome containing piperine (AUC0–∞) compared to that of raw PPD (AUC0–∞) were 166% and 248%, respectively.
Conclusion
The increased bioavailability of PPD-cubosome loaded with piperine is due to an increase in absorption and inhibition of metabolism of PPD by cubic nanoparticles containing piperine rather than because of improved release of PPD. The cubic nanoparticles containing piperine may be a promising oral carrier for anticancer drugs with poor oral absorption and that undergo extensive metabolism by cytochrome P450.
doi:10.2147/IJN.S38203
PMCID: PMC3576041  PMID: 23426652
20(S)-protopanaxadiol; cubosome; piperine; Caco-2 cell monolayer; bioavailability; metabolites
2.  Preparation of a nanoscale baohuoside I-phospholipid complex and determination of its absorption: in vivo and in vitro evaluations 
Background
Baohuoside I is a potential anticancer drug for a variety of malignancies and has been approved for in vitro use. However, baohuoside I has very poor oral absorption.
Methods
In the present study, we prepared baohuoside I-phospholipid complexes of different diameters and determined their physicochemical properties using transmission electron microscopy, ultraviolet spectroscopy, and differential scanning calorimetry. The in vitro absorption of baohuoside I and baohuoside I-phospholipid complexes of different sizes were compared using the Caco-2 cell culture model, and subsequently, the bioavailability of baohuosidel and its complexes were estimated in vivo.
Results
Compared with the large-sized phospholipid complexes, a nanoscale phospholipid complex improved the oral bioavailability of baohuoside I. In addition, our results suggest that the smaller the particle size, the faster the complexes crossed the Caco-2 monolayer and the faster they were resorbed after oral administration in rats. The relative oral bioavailability of a nanoscale size 81 ± 10 nm baohuoside I-phospholipid complex (area under the concentration-time curve [AUC]0–∞) was 342%, while that of baohuoside I and a 227.3 ± 65.2 μm baohuoside I-phospholipid complex was 165%.
Conclusion
We enhanced the oral bioavailability of baohuoside I by reducing the particle size of the phospholipid complex to the nanometer range, thereby improving its potential for clinical application.
doi:10.2147/IJN.S35965
PMCID: PMC3446837  PMID: 23028219
nanoscale phospholipid complex; Caco-2 cell monolayer; bioavailability; oral absorption
3.  The Clinical Significance and Risk Factors of Solitary Lymph Node Metastasis in Gastric Cancer 
PLoS ONE  2015;10(1):e0114939.
Aims
To assess the clinical significance and risk factors of solitary lymph node metastasis (SLM) in gastric carcinoma and establish a more accurate method to evaluate the possibility of lymph node metastasis (LM).
Methods
A total of 385 patients with gastric carcinoma who underwent D2 lymphadenectomy at the Cancer Center of Sun Yat-Sen University were included in this research. Then we used a group of data from Sun Yat-sen University Gastrointestinal Hospital (SYSUGIH) to validate the accuracy of our developed method. The χ2 test, Kaplan–Meier analysis, log-rank test, COX model, and discriminate analysis were used to analyze the data with SPSS13.0.
Results
We found that the LM number and pathological T staging were independent prognostic risk factors. CEA grading, LN status by CT, and T staging by CT were independent risk factors for LM in gastric carcinoma. In addition, we developed the equation Y = -5.0 + X1 + 1.8X3 + 0.7X4 (X1 = CEA grading, X3 = LN status by CT, X4 = T staging by CT) to evaluate the situation of LM. The data from SYSUGIH shows this equation has a better accuracy compared with CT.
Conclusions
SLM is an independent risk factor in gastric cancer. And there was no survival difference between the skip metastasis group and the other SLM group (P = 0.659). It is inappropriate for the patient with SLM doing a standard D2 lymphadenectomy, due to the fact that LM rarely occurs in the splenic artery, splenic hilum. The risk factors for LM include CEA grading, LN status by CT, and T staging by CT. And we can use Y = -5.0 + X1 + 1.8X3 + 0.7X4 (X1, CEA grading, X3 = LN status by CT, X4 = T staging by CT, the critical value is 0.3) to estimate the possibility of LM, which has a better accuracy compared with CT.
doi:10.1371/journal.pone.0114939
PMCID: PMC4310611  PMID: 25633364
4.  Efficacy of Continuous Positive Airway Pressure on Testosterone in Men with Obstructive Sleep Apnea: A Meta-Analysis 
PLoS ONE  2014;9(12):e115033.
Objective
To evaluate the efficacy of continuous positive airway pressure (CPAP) on serum testosterone in men with obstructive sleep apnea (OSA).
Methods
Two reviewers independently searched PubMed, Cochrane library, Embase and Web of Science before June 2014. Information on characteristics of subjects, study design, pre- and post-CPAP treatment of serum total testosterone, free testosterone and sexual hormone blinding protein (SHBG) was extracted for analysis.
Results
A total of 7 studies with 9 cohorts that included 232 men were pooled into meta-analysis. There was no change of total testosterone levels before and after CPAP treatment in OSA men (standardized mean difference (SMD) = −0.14, 95%CI: −0.63 to 0.34, z = 0.59, p = 0.558), even subdivided by CPAP therapeutic duration (>3 months). Meanwhile, no significant differences in free testosterone and SHBG were detected after CPAP treatment (SMD =  0.16, 95%CI: −0.09 to 0.40, z = 1.25, p = 0.211 and SMD = −0.58, 95%CI: −1.30 to 0.14, z = 1.59, p = 0.112, respectively).
Conclusion
CPAP has no influence on testosterone levels in men with OSA, further large-scale, well-design interventional investigation is needed.
doi:10.1371/journal.pone.0115033
PMCID: PMC4263732  PMID: 25503098
5.  Treatment of giant emphysamous bulla with endobronchial valves in patients with chronic obstructive pulmonary disease: a case series 
Journal of Thoracic Disease  2014;6(12):1674-1680.
Background
Giant emphysamtous bulla (GEB) can negatively affect the pulmonary functions of chronic obstructive pulmonary diseases (COPD) patients, including decreased forced expiratory volume in 1 s (FEV1) and increased functional residual capacity (FRC). The aim of this study was to evaluate the efficacy of endobronchial valve (EBV) to treat bullae and to find efficacy predictors of successful treatment.
Methods
Five COPD patients with giant bulla were treated using EBVs. Before the EBV deployment, collateral ventilation (CV) between the targeted and adjacent lobes was evaluated with Chartis system.
Results
In the two patients with negative CV, the mean value of FEV1 increased from 27.1±11.4% of predicted value before EBV treatment to 32.8±12.0% (P>0.05) at 1 month after EBV treatment, than to 31.7±24.5% (P>0.05) at 6 months after EBV treatment. Only one patient, whose bulla occupied the whole right middle lung, displayed sustained improvement of FEV1 at 6 months after EBV treatment. In the three patients with positive CV, the mean value of FEV1 decreased from 28.8±19.0% of predicted value before EBV treatment to 24.8±12.6% (P>0.05) at 1 month after EBV treatment, than to 22.1±10.8% (P>0.05) at 6 months after EBV treatment.
Conclusions
EBV is an effective measure to treat highly selected COPD patients with giant bulla. Although, EBV treatment can achieve transient improvement of lung function at patients with CV negative bulla, long-term benefit was merely observed at the patient with a bulla at right middle lobe (RML).
doi:10.3978/j.issn.2072-1439.2014.11.07
PMCID: PMC4283300  PMID: 25589959
Endobronchial valve (EBV); bulla; chronic obstructive pulmonary diseases (COPD); collateral ventilation (CV)
6.  Identification and Evaluation of Reliable Reference Genes for Quantitative Real-Time PCR Analysis in Tea Plant (Camellia sinensis (L.) O. Kuntze) 
Reliable reference selection for the accurate quantification of gene expression under various experimental conditions is a crucial step in qRT-PCR normalization. To date, only a few housekeeping genes have been identified and used as reference genes in tea plant. The validity of those reference genes are not clear since their expression stabilities have not been rigorously examined. To identify more appropriate reference genes for qRT-PCR studies on tea plant, we examined the expression stability of 11 candidate reference genes from three different sources: the orthologs of Arabidopsis traditional reference genes and stably expressed genes identified from whole-genome GeneChip studies, together with three housekeeping gene commonly used in tea plant research. We evaluated the transcript levels of these genes in 94 experimental samples. The expression stabilities of these 11 genes were ranked using four different computation programs including geNorm, Normfinder, BestKeeper, and the comparative ∆CT method. Results showed that the three commonly used housekeeping genes of CsTUBULIN1, CsACINT1 and Cs18S rRNA1 together with CsUBQ1 were the most unstable genes in all sample ranking order. However, CsPTB1, CsEF1, CsSAND1, CsCLATHRIN1 and CsUBC1 were the top five appropriate reference genes for qRT-PCR analysis in complex experimental conditions.
doi:10.3390/ijms151222155
PMCID: PMC4284700  PMID: 25474086
Camellia sinensis; reference gene; qRT-PCR; tea plant; gene expression; normalization
7.  Thoracolumbar fracture dislocations treated by posterior reduction, interbody fusion and segmental instrumentation 
Indian Journal of Orthopaedics  2014;48(6):568-573.
Background:
Literature describing the application of modern segmental instrumentation to thoracic and lumbar fracture dislocation injuries is limited and the ideal surgical strategy for this severe trauma remains controversial. The purpose of this article was to investigate the feasibility and efficacy of single-stage posterior reduction with segmental instrumentation and interbody fusion to treat this type of injury.
Materials and Methods:
A retrospective review of 30 patients who had sustained fracture dislocation of the spine and underwent single stage posterior surgery between January 2007 and December 2011 was performed. All the patients underwent single stage posterior pedicle screw fixation, decompression and interbody fusion. Demographic data, medical records and radiographic images were reviewed thoroughly.
Results:
Ten females and 20 males with a mean age of 39.5 years were included in this study. Based on the AO classification, 13 cases were Type B1, 4 cases were B2, 4 were C1, 6 were C2 and 3 cases were C3. The average time of the surgical procedure was 220 min and the average blood loss was 550 mL. All of the patients were followed up for at least 2 years, with an average of 38 months. The mean preoperative kyphosis was 14.4° and reduced to -1.1° postoperatively. At the final followup, the mean kyphosis was 0.2°. The loss of correction was small (1.3°) with no significant difference compared to postoperative kyphotic angle (P = 0.069). Twenty seven patients (90%) achieved definitive bone fusion on X-ray or computed tomography imaging within 1 year followup. The other three patients were suspected possible pseudarthrosis. They remained asymptomatic without hardware failure or local pain at the last followup.
Conclusion:
Single stage posterior reduction using segmental pedicle screw instrumentation, combined with decompression and interbody fusion for the treatment of thoracic or lumbar fracture-dislocations is a safe, less traumatic and reliable technique. This procedure can achieve effective reduction, sagittal angle correction and solid fusion.
doi:10.4103/0019-5413.144219
PMCID: PMC4232825  PMID: 25404768
Fracture dislocation; posterior interbody fusion; segmental instrumentation; spine trauma; thoracolumbar spine; Spine; spinal fractures; dislocations; spinal fusion; instrumentation
8.  Five New Secondary Metabolites Produced by a Marine-Associated Fungus, Daldinia eschscholzii 
Marine Drugs  2014;12(11):5563-5575.
Five new compounds, including a benzopyran ribonic glycoside, daldiniside A (1), two isocoumarin ribonic glycosides, daldinisides B (2) and C (3), and two alkaloids, 1-(3-indolyl)-2R,3-dihydroxypropan-1-one (4) and 3-ethyl-2,5-pyrazinedipropanoic acid (5), along with five known compounds (6–10), were isolated from the EtOAc extract of the marine-associated fungus, Daldinia eschscholzii. Their structures were elucidated by extensive physicochemical and spectroscopic properties, besides comparison with literature data. The absolute configurations of compounds 1–3 were corroborated by chemical transformation, GC analysis and X-ray crystallographic analysis. Meanwhile, the absolute configuration of compound 4 and the planar structure of compound 6 were also determined based on the X-ray diffraction analysis. The cytotoxicity of compounds 1–10, antifungal and anti-HIV activities of compounds 1–5 and the in vitro assay for glucose consumption of compounds 1–3 were done in the anti-diabetic model, whereas none showed obvious activity.
doi:10.3390/md12115563
PMCID: PMC4245545  PMID: 25419997
marine-associated fungus; Daldinia eschscholzii; secondary metabolites; hydrolysis; GC analysis; X-ray diffraction analysis
9.  Hederagenin from the leaves of ivy (Hedera helix L.) induces apoptosis in human LoVo colon cells through the mitochondrial pathway 
Background
Colorectal cancer has become one of the leading cause of cancer morbidity and mortality throughout world. Hederagenin, a derivative of oleanolic acid isolated from the leaves of ivy (Hedera helix L.), has been shown to have potential anti-tumor activity. The study was conducted to evaluate whether hederagenin could induce apoptosis of human colon cancer LoVo cells and explore the possible mechanism.
Methods
MTT assay was used for evaluating cell viability while Annexin V-FITC/PI assay and Hoechst 33342 nuclear stainining were used for the determination of apoptosis and mitochondrial membrane potential. DCFH-DA fluorescence staining and flow cytometry were used to measure ROS generation. Real-time PCR and western blot analysis were performed for apoptosis-related protein expressions.
Results
MTT assay showed that hederagenin could significantly inhibit the viability of LoVo cells in a concentration-dependent and time-dependent manner by IC50 of 1.39 μM at 24 h and 1.17 μM at 48 h. The apoptosis ratio was significantly increased to 32.46% and 81.78% by the induction of hederagenin (1 and 2 μM) in Annexin V-FITC/PI assay. Hederagenin could also induce the nuclear changes characteristic of apoptosis by Hoechst 33342 nuclear stainining under fluorescence microscopy. DCFH-DA fluorescence staining and flow cytometry showed that hederagenin could increase significantly ROS generation in LoVo cells. Real-time PCR showed that hederagenin induced the up-regulation of Bax and down-regulation of Bcl-2, Bcl-xL and Survivin. Western blotting analysis showed that hederagenin decreased the expressions of apoptosis-associated proteins Bcl-2, procaspase-9, procaspase-3, and polyADP- ribosepolymerase (PARP) were increased, while the expressions of Bax, caspase-3, caspase-9 were increased. However, there was no significant change on caspase-8.
Conclusions
These results indicated that the disruption of mitochondrial membrane potential might contribute to the apoptosis of hederagenin in LoVo cells. Our findings suggested that hederagenin might be a promising therapeutic candidate for human colon cancer.
doi:10.1186/1472-6882-14-412
PMCID: PMC4216349  PMID: 25342273
Hederagenin; Apoptosi; Mitochondrial pathway; Colon cancer
10.  Efficacy of Surgical Resection for Pulmonary Metastases from Hepatocellular Carcinoma 
Background
The lung is one of the most common sites for extrahepatic metastasis from hepatocellular carcinoma (HCC). This study aimed to assess the efficacy of surgical resection for pulmonary metastases from HCC.
Material/Methods
The medical records of 9 patients who underwent pulmonary metastasectomy from HCC at 2 institutions were retrospectively studied, together with a review of studies reporting the outcomes of at least 5 patients in the Chinese and English languages.
Results
There were no perioperative deaths or major complications. The 1-, 3-, and 5-year overall survival rate after surgery was 100%, 44.4%, and 33.3%, respectively. A total of 19 studies involving 443 patients who underwent pulmonary metastasectomy for metastasis of HCC were included in the literature review. The median mortality rate was 0% (range, 0–7.1%). The median survival ranged from 10.7 to 77 (median=33.2) months, and the 5-year overall survival rate ranged from 11.5% to 75% (median=36%).
Conclusions
Surgical resection is a safe and effective treatment in selected patients with pulmonary metastases from HCC.
doi:10.12659/MSM.890853
PMCID: PMC4156333  PMID: 25172966
Carcinoma; Hepatocellular; Metastasectomy; Recurrence
11.  miR-30b, Down-Regulated in Gastric Cancer, Promotes Apoptosis and Suppresses Tumor Growth by Targeting Plasminogen Activator Inhibitor-1 
PLoS ONE  2014;9(8):e106049.
Background
Gastric cancer is one of the most common malignant diseases worldwide. Emerging evidence has shown that microRNAs (miRNAs) are associated with tumor development and progression. Our previous studies have revealed that H. pylori infection was able to induce the altered expression of miR-30b in gastric epithelial cells. However, little is known about the potential role of miR-30b in gastric cancer.
Methods
We analyzed the expression of miR-30b in gastric cancer cell lines and human gastric cancer tissues. We examined the effect of miR-30b mimics on the apoptosis of gastric cancer cells in vitro by flow cytometry (FCM) and caspase-3/7 activity assays. Nude mouse xenograft model was used to determine whether miR-30b is involved in tumorigenesis of gastric cancer. The target of miR-30b was identified by bioinformatics analysis, luciferase assay and Western blot. Finally, we performed the correlation analysis between miR-30b and its target expression in gastric cancer.
Results
miR-30b was significantly down-regulated in gastric cancer cells and human gastric cancer tissues. Enforced expression of miR-30b promoted the apoptosis of gastric cancer cells in vitro, and miR-30b could significantly inhibit tumorigenicity of gastric cancer by increasing the apoptosis proportion of cancer cells in vivo. Moreover, plasminogen activator inhibitor-1 (PAI-1) was identified as the potential target of miR-30b, and miR-30b level was inversely correlated with PAI-1 expression in gastric cancer. In addition, silencing of PAI-1 was able to phenocopy the effect of miR-30b overexpression on apoptosis regulation of cancer cells, and overexpression of PAI-1 could suppressed the effect of promoting cell apoptosis by miR-30b, indicating PAI-1 is potentially involved in miR-30b-induced apoptosis on cancer cells.
Conclusion
miR-30b may function as a novel tumor suppressor gene in gastric cancer by targeting PAI-1 and regulating the apoptosis of cancer cells. miR-30b could serve as a potential biomarker and therapeutic target against gastric cancer.
doi:10.1371/journal.pone.0106049
PMCID: PMC4149503  PMID: 25170877
12.  Association of the Three Common SNPs of Cyclooxygenase-2 Gene (rs20417, rs689466, and rs5275) with the Susceptibility of Breast Cancer: An Updated Meta-Analysis Involving 34,590 Subjects 
Disease Markers  2014;2014:484729.
Several single nucleotide polymorphisms have been identified in cyclooxygenase-2 (COX-2) genes (e.g., −765 G>C (rs20417), −1195G>A (rs689466), and 8473 C>T (rs5275)). The association of these SNPs with the risk of different cancer types is still controversial. This study aims to evaluate the correlation between these SNPs and breast cancer risk in different ethnic groups. We have searched PubMed, Web of Knowledge, and Embase for relevant studies. Odds ratios (ORs) with 95% confidence intervals (CIs) were used to estimate the strength of the associations. A total of 13 studies (15,330 cases and 19,260 controls) were eligible for meta-analysis. This meta-analysis showed that COX-2 rs20417 polymorphism was correlated with an increased risk of breast cancer in Caucasians, while rs689466 was associated with a decreased risk of breast cancer in Caucasians. The rs5275 polymorphism had no association with breast cancer risk.
doi:10.1155/2014/484729
PMCID: PMC4151597  PMID: 25214704
13.  miR-148b functions as a tumor suppressor in non-small cell lung cancer by targeting carcinoembryonic antigen (CEA) 
Aberrant expression of miR-148b has been found in several types of cancer, but its expression and potential biologic role in non-small cell lung cancer (NSCLC) are still largely unknown. Here, we found that miR-148b was commonly under-expressed in human non-small celllung cancer (NSCLC) specimens and cell lines. The overexpression of miR-148b dramatically suppressed NSCLC cell proliferation and migration. Furthermore, miR-148b could regulate carcinoembryonic antigen (CEA) expression by luciferase reporter assay. On the other hand, CEA was widely up-regulated in NSCLC specimens, and its mRNA levels were inversely correlated with miR-148b expression. These suggest that CEA expression may be regulated by miR-148b. Collectively, our findings indicate miR-148b is low expression in NSCLC cells, which results in CEA overexpression and disease progression in NSCLC patients.
PMCID: PMC4161539  PMID: 25232379
NSCLC; CEA; miR-148b; migration; tumor suppressors
14.  Human leukocyte antigen-DRB1*1501 and DQB1*0602 alleles are cervical cancer protective factors among Uighur and Han people in Xinjiang, China 
Human papillomavirus (HPV) infection is a major risk factor for cervical cancer. However, only some high risk human papillomavirus (HR-HPV)-infected women progress to cervical cancer, host immunogenetic factors human leukocyte antigen (HLA) may account for viral antigens presenting individually or together in the progression to cervical cancer. This study examined the association between the development of invasive cervical cancer (ICC) and the determinant factors including HLA-DRB1*1501 and DQB1*0602, HR-HPV infection among Chinese Uighur and Han populations. Blood samples, cervical swabs and biopsies were obtained from 287 patients with ICC (192 Uighurs and 95 Hans) and 312 healthy controls (218 Uighurs and 94 Hans). HPV DNA was detected by PCR and HLA-DRB1*1501 and DQB1*0602 alleles were performed using PCR-SSP method. HPV16 infection rates was significantly higher among Uighur and Han with ICC as compared to healthy controls (OR = 58.317; 95% CI: 39.663-85.744; OR = 33.778; 95% CI: 12.581-90.691; P < 0.05 for all). HLA-DRB1*1501 (OR = 0.305; 95% CI: 0.115-0.813; P < 0.05) and HLA-DRB1*1501-DQB1*0602 haplotype frequencies (OR = 0.274; 95% CI: 0.086-0.874; P < 0.05) were significantly reduced in Han ICC. The HLA-DQB1*0602 frequency significantly decreased among Uighur women with ICC (OR = 0.482; 95% CI: 0.325-0.716; P < 0.05). Similar tendencies were observed for DQB1*0602 with HPV16-positive ICC (OR = 0.550; 95% CI: 0.362-0.837; P < 0.05). This study suggests that HLA-DRB1*1501 and DQB1*0602 alleles may influence the immune response to HPV16 infection and decrease the risk of ICC among Uighurs and Hans in Xinjiang, China.
PMCID: PMC4203236  PMID: 25337265
HLA; invasive cervical cancer; HPV; susceptibility; Uighur
15.  Intraocular Pressure and Estimated Cerebrospinal Fluid Pressure. The Beijing Eye Study 2011 
PLoS ONE  2014;9(8):e104267.
Purpose
To examine a potential association between intraocular pressure (IOP) and cerebrospinal fluid pressure (CSFP) in a population-based setting.
Methods
The population-based Beijing Eye Study 2011 included 3468 individuals with a mean age of 64.6±9.8 years (range: 50–93 years). A detailed ophthalmic examination was performed. Based on a previous study with lumbar cerebrospinal fluid pressure (CSFP) measurements, CSFP was calculated as CSFP [mm Hg] = 0.44×Body Mass Index [kg/m2]+0.16×Diastolic Blood Pressure [mm Hg]–0.18×Age [Years].
Results
In multivariate analysis, IOP was associated with higher estimated CSFP (P<0.001; standardized correlation coefficient beta: 0.27; regression coefficient B: 0.20; 95% confidence interval (CI): 0.16, 0.24), after adjusting for thinner central corneal thickness (P<0.001; beta: 0.45; B: 0.04;95%CI: 0.04,0.04), smaller corneal curvature radius (P<0.001; beta:−0.11; B:−1.13;95%CI:−1.61,−0.64), shallower anterior chamber depth (P = 0.01; beta:−0.05; B:−0.33;95%CI:−0.59,−0.08) and longer axial length (P = 0.002; beta: 0.08; B: 0.20;95%CI: 0.08,0.32)), and after adjusting for the systemic parameters of higher pulse rate (P<0.001; beta: 0.08; B: 0.02;95%CI: 0.01,0.03), higher prevalence of arterial hypertension (P = 0.002; beta: 0.06; B: 0.32;95%CI: 0.12,0.53)), frequency of drinking alcohol (P = 0.02; beta: 0.04; B: 0.09;95%CI: 0.01,0.17), higher blood concentration of triglycerides (P = 0.001; beta: 0.06; B: 0.06;95%CI: 0.02,0.10) and cholesterol (P = 0.049; beta: 0.04; B: 0.08;95%CI: 0.00,0.17), and body mass index (P<0.001; beta:−0.13; B:−0.09;95%CI:−0.13,−0.06). In a parallel manner, estimated CSFP (mean: 10.8±3.7 mm Hg) was significantly associated with higher IOP (P<0.001; beta: 0.13; B: 0.18;95%CI: 0.13,0.23) after adjusting for rural region of habitation (P<0.001; beta:−0.37; B:−2.78;95%CI:−3.07,−2.48), higher systolic blood pressure (P<0.001; beta: 0.34; B: 0.06;95%CI: 0.05,0.07), higher pulse rate (P = 0.003; beta: 0.05; B: 0.02;95%CI: 0.01,0.03), taller body height (P<0.001; beta: 0.11; B: 0.05;95%CI: 0.03,0.07), higher blood concentration of cholesterol (P = 0.003; beta: 0.05; B: 0.17;95%CI: 0.06,0.28) and higher level of education (P = 0.003; beta: 0.09; B: 0.30;95%CI: 0.16,0.45).
Conclusions
IOP was positively associated with estimated CSFP after adjusting for other ocular and systemic parameters. As a corollary, higher estimated CSFP was significantly associated with higher IOP in multivariate analysis. It fits with the notion that the arterial blood pressure, estimated CSFP and IOP are physiologically correlated with each other.
doi:10.1371/journal.pone.0104267
PMCID: PMC4126772  PMID: 25105777
17.  The complete mitochondrial genome of the scab mite Psoroptes cuniculi (Arthropoda: Arachnida) provides insights into Acari phylogeny 
Parasites & Vectors  2014;7:340.
Background
Limited available sequence information has greatly impeded population genetics, phylogenetics and systematics studies in the subclass Acari (mites and ticks). Mitochondrial (mt) DNA is well known to provide genetic markers for investigations in these areas, but complete mt genomic data have been lacking for many Acari species. Herein, we present the complete mt genome of the scab mite Psoroptes cuniculi.
Methods
P. cuniculi was collected from a naturally infected New Zealand white rabbit from China and identified by morphological criteria. The complete mt genome of P. cuniculi was amplified by PCR and then sequenced. The relationships of this scab mite with selected members of the Acari were assessed by phylogenetic analysis of concatenated amino acid sequence datasets by Bayesian inference (BI), maximum likelihood (ML) and maximum parsimony (MP).
Results
This mt genome (14,247 bp) is circular and consists of 37 genes, including 13 genes for proteins, 22 genes for tRNA, 2 genes for rRNA. The gene arrangement in mt genome of P. cuniculi is the same as those of Dermatophagoides farinae (Pyroglyphidae) and Aleuroglyphus ovatus (Acaridae), but distinct from those of Steganacarus magnus (Steganacaridae) and Panonychus citri (Tetranychidae). Phylogenetic analyses using concatenated amino acid sequences of 12 protein-coding genes, with three different computational algorithms (BI, ML and MP), showed the division of subclass Acari into two superorders, supported the monophylies of the both superorders Parasitiformes and Acariformes; and the three orders Ixodida and Mesostigmata and Astigmata, but rejected the monophyly of the order Prostigmata.
Conclusions
The mt genome of P. cuniculi represents the first mt genome of any member of the family Psoroptidae. Analysis of mt genome sequences in the present study has provided new insights into the phylogenetic relationships among several major lineages of Acari species.
doi:10.1186/1756-3305-7-340
PMCID: PMC4223567  PMID: 25052180
Psoroptes cuniculi; Mitochondrial genome; Mitochondrial DNA; Phylogenetic analyses
18.  Effect of emodin on the cariogenic properties of Streptococcus mutans and the development of caries in rats 
Emodin is an active herbal component traditionally used in East Asian countries for treating a variety of diseases. The present study investigated the effects of emodin on specific virulence factors of Streptococcus mutans (S. mutans) in vitro and on caries development in vivo. The growth and acid production of S. mutans were significantly inhibited by emodin (0.5–2 mg/ml). Emodin also significantly suppressed the synthesis of insoluble glucans by S. mutans. Furthermore, the topical application of emodin reduced the incidence and severity of carious lesions in rats. These results suggest that the natural compound emodin may be a novel pharmacological agent for the prevention and treatment of dental caries.
doi:10.3892/etm.2014.1857
PMCID: PMC4151691  PMID: 25187845
caries; Streptococcus mutans; glucosyltransferase; emodin
19.  Identification of miR-140-3p as a marker associated with poor prognosis in spinal chordoma 
Objective: To investigate the expression profile of miR-140-3p in formalin-fixed paraffin-embedded (FFPE) tissues of spinal chordoma, and its correlation with the prognosis of spinal chordoma patients. Methods: Dysregulated miRNAs in FFPE tissues of spinal chordoma were identified by microarray analysis. MiR-140-3p expression in surgically removed spinal chordoma tissues of 42 spinal chordoma patients (27 males and 15 females, aged 29-76 years) and corresponding nucleus pulposus tissues of 14 patients with disc herniation as the healthy control group (8 males and 6 females, aged 24-73 years) was measured by real-time quantitative RT-PCR assay. The association of miR-140-3p expression with clinicopathologic characteristics of spinal chordoma patients was analyzed. Additionally, we investigated the prognostic significance of miR-140-3p with the use of Kaplan-Meier methods and a Cox proportional hazard model. Results: The expression of miR-140-3p was significantly higher in chordoma tissues than nucleus pulposus tissues (t = 3.530, P = 0.001). The expression of miR-140-3p positively correlated with surrounding muscle invasion. The Kapan-Meier survival analysis showed that the patients with high miR-140-3p expression had a significantly worse recurrence-free survival than those with a low expression (χ 2 = 31.270, P = 0.000, log-rank test). In addition, univariate and multivariate analyses for recurrence-free survival showed that miR-140-3p expression was an independent prognostic factor for patients with spinal chordoma (HR = 1.361, 95% CI: 1.135-1.633, P = 0.001). Conclusion: Over-expression of miR-140-3p is correlated with recurrence and tumor invasion, suggesting that miR-140-3p could be a new predictor for recurrence and prognosis in patients with spinal chordoma.
PMCID: PMC4152048  PMID: 25197358
MicroRNA; chordoma; spine; prognosis; recurrence; case-control study
20.  Descriptions of three new species of the Termitophilous tribe Termitopaediini in China (Coleoptera, Staphylinidae, Aleocharinae) 
ZooKeys  2014;91-100.
Three new species belonging to two genera of the aleocharine tribe Termitopaediini Seevers, viz., Dioxeuta rara Song & Li, sp. n., D. yunnanensis Song & Li, sp. n., and Termitopulex sinensis Song & Li, sp. n. from Baihualing Natural Reserve (Southwest China: Yunnan) are described and illustrated. This is the first record of Termitopulex Fauvel, 1899 from China.
doi:10.3897/zookeys.424.7670
PMCID: PMC4106102  PMID: 25061397
Termitopaediini; Dioxeuta; Termitopulex; termitophily; taxonomy; new species; China; Oriental region
21.  Qualitative and quantitative analysis of the major constituents in traditional Chinese medicine Danmu injection using LC-ESI-MSn and LC-DAD 
Pharmacognosy Magazine  2014;10(39):254-264.
Background:
Danmu injection, a traditional Chinese medicine (TCM) preparation made from Nauclea officinalis, has been commonly used for the treatment of cold, fever, swelling of throat in China. However, the chemical constituents in Danmu injection have not been clarified yet.
Objective:
a HPLC/DAD/ESI-MSn method was developed for qualitative and quantitative analysis of the components in Danmu injection.
Materials and Methods:
The chromatographic separation was performed on a Welch Material XB-C18 (4.6mm × 250mm, 5μm) using gradient elution with acetonitrile (A) and water containing 0.1% formic acid (B) as mobile phase at a flow rate of 1.0 ml/min.
Results:
Twenty-five compounds, including phenolic acid and phenol glycoside, iridous glycoside and glycoalkaloid were identified or tentatively deduced on the base of their retention behaviors, UV absorption, MS and MSn data with those elucidated references or literature. In addition, eleven compounds were simultaneously determined by HPLC–DAD, which was validated and successfully applied for determination of major components in Danmu injection.
Conclusion:
The results suggested that the established qualitative and quantitative method would be a powerful and reliable analytical tool for the characterization of multi-constituent in complex chemical system and quality control of Danmu injection.
doi:10.4103/0973-1296.137365
PMCID: PMC4159918  PMID: 25210312
Danmu injection; HPLC-DAD-ESI-MSn; major constituents; Nauclea officinalis; quality control
22.  Enhanced dissolution and stability of Tanshinone IIA base by solid dispersion system with nano-hydroxyapatite 
Pharmacognosy Magazine  2014;10(39):332-337.
Background:
Tanshinone IIA (TSIIA) exhibits a variety of cardiovascular effects; however, it has low solubility in water. The preparation of poorly soluble drugs for oral delivery is one of the greatest challenges in the field of formulation research. Among the approaches available, solid dispersion (SD) technique has proven to be one of the most commonly used these methods for improving dissolution and bioavailability of drugs, because of its relative simplicity and economy in terms of both preparation and evaluation.
Objective:
This study was aimed at investigating the dissolution behavior and physical stability of SDs of TSIIA by employing nano-hydroxyapatite (n-HAp).
Materials and Methods:
The TSIIA SDs was prepared to use a spray-drying method. First, an in vitro dissolution test was performed to assess dissolution characteristics. Next, a set of complementary techniques (differential scanning calorimetry, scanning electron microscopy, X-ray powder diffraction, and Fourier transform infrared spectroscopy) was used to monitor the physicochemical properties of the SDs. The SDs was stored at 40°C/75% relative humidity for 6 months, after which their stability was assessed.
Results:
TSIIA dissolution remarkably improved because of the formulation of the SDs with n-HAp particles. Comparisons with the corresponding physical mixtures revealed changes in the SDs and explained the formation of the amorphous phase. In the stability test, virtually no time-dependent decrease was observed in either in vitro drug dissolution or drug content.
Conclusion:
SD formulation with n-HAp may be a promising approach for enhancing the dissolution and stability of TSIIA.
doi:10.4103/0973-1296.137375
PMCID: PMC4159928  PMID: 25210322
Dissolution; nano-hydroxyapatite; solid dispersion; stability; Tanshinone IIA
23.  Recurrent inflammatory myofibroblastic tumors harboring PIK3CA and KIT mutations 
Inflammatory myofibroblastic tumour (IMT) is a relatively rare soft tissue malignancy. It exhibits locally aggressive behavior with a tendency for local recurrence and rare metastasis, and rare recurrent IMTs may show histological progression. The genetic hallmark of IMT is ALK rearrangement from chromosome arm 2p, but gene mutations involved in IMT remain poorly understood. The aim of the present study was to perform a pairwise comparison of the gene mutations occurring in primary and recurrent IMT from the same patient. We conducted a high-throughput analysis of 238 known mutations of 19 oncogenes in pairwise comparison primary and recurrent samples from 2 patients of IMT using Sequenom MassARRAY technology. Our results revealed 2 mutations in 2 recurrent lesion samples, including one in exon 11 of the KIT gene, resulting in a T-C substitution at position 1727 (L576P), the recurrent sample underwent histologic progression with “pleomorphic undifferentiated sarcoma-like” transformation; the other mutation was in exon 19 of the phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA) gene, resulting in a G-A substitution at position 1624 (E542K). Moreover, no any mutation was found in the primary lesion samples from 2 patients. Our findings suggest that variable genome changes might be present in IMT, especially during the progression from a primary tumour to recurrence. To the best of our knowledge, no such longitudinal study of IMT has been undertaken previously.
PMCID: PMC4128978  PMID: 25120743
Inflammatory myofibroblastic tumour; gene mutation; recurrent tumour; MassARRAY
24.  A case of blastic plasmacytoid dendritic cell neoplasm with ecchymotic lesions on the whole body 
Blastic plasmacytoid dendritic cell neoplasm (BPDCN) derived from plasmacytoid dendritic cell precursors is a very rare, and characterized by cutaneous and bone marrow involvement and leukemic spread. The neoplasm presents with an aggressive behavior, and the clinical findings include cytopenia, particularly thrombocytopenia. The tumor cells are negative for antigens of T- and B- cell lines. However, these cells express CD4, CD56 and CD123, which are markers of plasmacytoid dendritic cells, and negative for Epstein-Barr virus (EBV). From this point of view, a 71-year-old man who was initially found to have a cutaneous mass on his face and thorax was reported here, and initially was diagnosed as “eczema”. The skin rashes then became aggravated on a trial of low dose topical corticosteroid for 2 months. According to skin biopsy, the tumor cells reveal an immature blastic appearance and positive for CD4 and CD56, negative for CD3, CD20, indicating a diagnosis of BPDCN. Here, we report the dismal course of a patient with BPDCN without accepting further therapy, and only survived 3 months.
PMCID: PMC4129059  PMID: 25120824
Blastic plasmacytoid dendritic cell neoplasm; BPDCN; neoplasm
25.  Postsynaptic potentiation of corticospinal projecting neurons in the anterior cingulate cortex after nerve injury 
Molecular Pain  2014;10:33.
Long-term potentiation (LTP) is the key cellular mechanism for physiological learning and pathological chronic pain. In the anterior cingulate cortex (ACC), postsynaptic recruitment or modification of AMPA receptor (AMPAR) GluA1 contribute to the expression of LTP. Here we report that pyramidal cells in the deep layers of the ACC send direct descending projecting terminals to the dorsal horn of the spinal cord (lamina I-III). After peripheral nerve injury, these projection cells are activated, and postsynaptic excitatory responses of these descending projecting neurons were significantly enhanced. Newly recruited AMPARs contribute to the potentiated synaptic transmission of cingulate neurons. PKA-dependent phosphorylation of GluA1 is important, since enhanced synaptic transmission was abolished in GluA1 phosphorylation site serine-845 mutant mice. Our findings provide strong evidence that peripheral nerve injury induce long-term enhancement of cortical-spinal projecting cells in the ACC. Direct top-down projection system provides rapid and profound modulation of spinal sensory transmission, including painful information. Inhibiting cortical top-down descending facilitation may serve as a novel target for treating neuropathic pain.
doi:10.1186/1744-8069-10-33
PMCID: PMC4060852  PMID: 24890933

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