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To improve the availability of inorganic phosphorus (P) in soil, we investigated the role of three macromolecular organic acids (MOAs), including fulvic acid (FA), polyaspartic acid (PA), and tannic acid (TA), in reducing the fixation of inorganic P fertilizer in the soil. AlPO(4), FePO(4), and Ca(8)H(2)(PO(4))(6)·5H(2)O crystals were chosen as insoluble phosphate representatives in the soil to simulate the solubilization process of inorganic P by MOAs. The microstructural and physicochemical properties of AlPO(4), FePO(4), and Ca(8)H(2)(PO(4))(6)·5H(2)O were determined by scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), and X-ray photoelectron spectroscopy (XPS) before and after treatment of MOAs. In addition, the amounts of leached P and fixed inorganic P in Inceptisols and Alfisols affected by MOAs combined with superphosphate (SP) fertilizer were determined by soil leaching experiments. The presence of the three MOAs significantly increased the concentration of leached P and reduced the contents of insoluble inorganic phosphate formed with iron, aluminum, and calcium fixed in the soil, in which PA combined with SP had the most significant effect. Furthermore, the less inorganic P fixation in the combination treatment of MOAs and SP resulted in a greater wheat yield and P uptake. Therefore, MOAs could be a synergistic material for increasing P fertilizer utilization.

The purpose of the retrospective case–control study was to identify the causes of and risk factors for unplanned return to the operating room (uROR) within 24 hours in surgical patients. We examined 275 cases of 24-hour uROR in our hospital from January 2010 to December 2018. The reasons for 24-hour uROR were classified into several categories. Controls were randomly matched to cases in a 1:1 ratio with the selection criteria set for the same surgeon and operation code in the same corresponding year. The mortality rate was significantly higher in patients with 24-hour uROR (11.63% vs 5.23%). Bleeding was the most common etiology (172/275; 62.55%) and technical error (14.5%) also contributed to 24-hour uROR. The clinical factors that led to bleeding included a history of liver disease (P = .032), smoking (P = .002), low platelet count in preoperative screening (P = .012), and preoperative administration of antiplatelet or anticoagulant agents (P = .014). Clinicians should recognize the risk factors for bleeding and minimize errors to avoid the increase in patient morbidity and mortality that is associated with 24-hour uROR. Level of Evidence: Level IV.

Rationale: Despite the development of human papillomavirus vaccines and significant improvement in cervical cancer screening over the past few years, cervical cancer remains the fourth most common cancer in women of childbearing age after breast cancer, melanoma, and thyroid cancer. Patient concerns: In this case report, the patients are all cervical cancer with stage IB2 and IB3 during pregnancy, the management constitutes a major medical challenge related to the impact of treatment on both maternal and fetal outcomes. Neoadjuvant chemotherapy (NACT) is an innovative option for cervical cancer patients with stage IB2 and IB3 before cesarean delivery and radical hysterectomy, and many chemotherapeutic agents are available, cisplatin plus paclitaxel yielded good maternal and fetal outcomes to the authors’ knowledge. Diagnoses: Masses were discovered in the cervix of 4 pregnant women with a history of vaginal bleeding. Biopsy examination of the masses revealed cervical carcinoma, which was staged in accordance with the International Federation of Gynecology and Obstetrics (i.e., FIGO) system. Interventions: The patients were treated with paclitaxel plus cisplatin, followed by cesarean delivery and radical hysterectomy. Outcomes: The 4 patients were treated successfully, with no recurrence during follow-up periods of 14 to 56 months, and all of the children were doing well with no anomalies. Lessons: Although further data are required, in pregnant women with invasive cervical cancer, NACT with cisplatin plus paclitaxel followed by cesarean delivery and radical hysterectomy was a practical treatment option.

East Asians are reportedly at high risk of anticoagulant-related bleeding; therefore, some physicians prefer to prescribe low-dose direct oral anticoagulants (DOACs). Little is known about the therapeutic effectiveness and safety of off-label reduced-dose apixaban in East Asians with nonvalvular atrial fibrillation (AF). We aimed to investigate the effectiveness and safety of off-label reduced-dose apixaban in Taiwanese patients with nonvalvular AF. This retrospective cohort study enrolled 1073 patients with nonvalvular AF who took apixaban between July 2014 and October 2018 from 4 medical centers in southern Taiwan. The primary outcomes included thromboembolic events (stroke/transient ischemic attack or systemic embolism), major bleeding, and all-cause mortality. Among all patients, 826 (77%) patients were classified as the “per-label adequate-dose” treatment group (i.e., consistent with the Food and Drug Administration label recommendations) while 247 (23%) patients were the “off-label reduced-dose” treatment group. The mean follow-up period was 17.5 ± 13 months. The “off-label reduced-dose” group did not have a lower major bleeding rate than the “per-label adequate-dose” group (4.8% vs 3.8%, adjusted hazard ratio [HR] 1.20, 95% confidence interval [CI] 0.69–2.09), but had a nonsignificantly higher incidence of thromboembolic events (4.23% vs 3.05%, adjusted HR: 1.29, 95% CI: 0.71–2.34). An off-label reduced-dose apixaban treatment strategy may not provide incremental benefits or safety for Taiwanese patients with nonvalvular AF.

A large number of chemokines, cytokines, other trophic factors and the extracellular matrix molecules form a favorable microenvironment for peripheral nerve regeneration. This microenvironment is one of the major factors for regenerative success. Therefore, it is important to investigate the key molecules and regulators affecting nerve regeneration after peripheral nerve injury. However, the identities of specific cytokines at various time points after sciatic nerve injury have not been determined. The study was performed by transecting the sciatic nerve to establish a model of peripheral nerve injury and to analyze, by protein microarray, the expression of different cytokines in the distal nerve after injury. Results showed a large number of cytokines were up-regulated at different time points post injury and several cytokines, e.g., ciliary neurotrophic factor, were downregulated. The construction of a protein-protein interaction network was used to screen how the proteins interacted with differentially expressed cytokines. Kyoto Encyclopedia of Genes and Genomes pathway and Gene ontology analyses indicated that the differentially expressed cytokines were significantly associated with chemokine signaling pathways, Janus kinase/signal transducers and activators of transcription, phosphoinositide 3-kinase/protein kinase B, and notch signaling pathway. The cytokines involved in inflammation, immune response and cell chemotaxis were up-regulated initially and the cytokines involved in neuronal apoptotic processes, cell-cell adhesion, and cell proliferation were up-regulated at 28 days after injury. Western blot analysis showed that the expression and changes of hepatocyte growth factor, glial cell line-derived neurotrophic factor and ciliary neurotrophic factor were consistent with the results of protein microarray analysis. The results provide a comprehensive understanding of changes in cytokine expression and changes in these cytokines and classical signaling pathways and biological functions during Wallerian degeneration, as well as a basis for potential treatments of peripheral nerve injury. The study was approved by the Institutional Animal Care and Use Committee of the Chinese PLA General Hospital, China (approval number: 2016-x9-07) in September 2016.

An outcome of building sustainable urban forests is that people’s well-being is improved when they are exposed to trees. Facial expressions directly represents one’s inner emotions, and can be used to assess real-time perception. The emergence and change in the facial expressions of forest visitors are an implicit process. As such, the reserved character of Asians requires an instrument rating to accurately recognize expressions. In this study, a dataset was established with 2,886 randomly photographed faces from visitors at a constructed urban forest park and at a promenade during summertime in Shenyang City, Northeast China. Six experts were invited to choose 160 photos in total with 20 images representing one of eight typical expressions: angry, contempt, disgusted, happy, neutral, sad, scared, and surprised. The FireFACE ver. 3.0 software was used to test hit-ratio validation as an accuracy measurement (ac.) to match machine-recognized photos with those identified by experts. According to the Kruskal-Wallis test on the difference from averaged scores in 20 recently published papers, contempt (ac. = 0.40%, P = 0.0038) and scared (ac. = 25.23%, P = 0.0018) expressions do not pass the validation test. Both happy and sad expression scores were higher in forests than in promenades, but there were no difference in net positive response (happy minus sad) between locations. Men had a higher happy score but lower disgusted score in forests than in promenades. Men also had a higher angry score in forests. We conclude that FireFACE can be used for analyzing facial expressions in Asian people within urban forests. Women are encouraged to visit urban forests rather than promenades to elicit more positive emotions.

Streetlamps enforce night lighting on urban forest trees, but scarce information is available concerning the ecophysiological performance of street trees under these conditions. In this study, maple (Acer truncatum Bunge) and oak (Quercus mongolica Fisch. ex Ledeb.) seedlings were cultured with simulated exposure to streetlamp spectra in white (red/green/blue, 7.7:1.0:2.2) and red plus blue (RB; red/green/blue, 4.6:0.0:1.0) lights with photosynthetic photon flux rate of 80 μmol m-2 s-1 in a 18-h photoperiod. Nitrogen (N) was loaded through 15 weekly applications to an amount of 80 mg N plant-1 to mimic the mineral N deposition to landscape trees. Variables of biomass, carbohydrate accumulation, N and water contents were rarely found difference between the two LED-spectra treatments for both species. Compared to the un-lighted control, the RB spectrum lowered N concentration in oak seedlings and water content in maple seedlings. The white light spectrum resulted in an increase of starch concentration. Carbohydrate concentration had a positive relationship with biomass and N content across two species but a negative relationship with water content in maple seedlings. Overall, streetlamp-lights imposed effects on tree growth by a prolonged photoperiod instead of specific spectrum. Maple had a strong response of water uptake to streetlamp lighting at the cost of carbohydrate consumption, but oak had scarce demand of water-use for growth.

Rationale: Follicular dendritic cell sarcoma (FDCS) is a rare malignant tumor derived from follicular dendritic cells, and is often associated with Castleman disease. Here we present a rare case of paraneoplastic pemphigus (PNP) with FDCS which required multidisciplinary approach for the diagnosis and treatment. Patient concerns: A 28-year-old Chinese female had FDCS recurrence, and primary clinical manifestation was PNP. Diagnoses: PNP with FDCS. Interventions: The patient received gamma globulin infusion, took anlotinib, and underwent plasma exchange therapy. Outcomes: The skin lesions recovered and there was no evidence of tumor recurrence. Lessons: The diagnosis and management of PNP with FDCS require close cooperation among surgeons, dermatologists, hematologists, otolaryngologists, oncologists, radiologists, pathologists, and respiratory doctors. The interesting clinical manifestations of this patient provide a multifaceted approach to the investigation of the interactions among FDCS, Castleman disease, and PNP.

To analyze risk factors associated with heterotopic pregnancy and the uterine pregnant outcome of those patients after surgery. We retrospectively analyzed 22 patients diagnosed as HP after in vitro fertilization (IVF) between January 2015 and December 2018. HP was diagnosed at gestation age of 55.4 ± 11.8 days. HP were presented as irregular vaginal bleeding, abdominal pain, and sometimes no symptoms. 81.8% of ectopic lesion in HP occurred at fallopian tubes, especially ampullary; cornual pregnancy takes up 13.6%. Compared with clinical intrauterine pregnancy (IUP), IVF with tubal infertility factors had higher risks of HP (OR 4.185, 95% CI 1.080– 16.217); IVF with pelvic adhesion also had higher risks of HP (OR 5.552 95% CI 1.677–18.382); IVF with more than 2 embryos transferred increased risks of HP (OR 23.253, 95% CI 1.804–299.767). The abortion rates of surgery-treated HP and IUP after IVF were 27.8% versus 10.3% (P = .042). These results demonstrate IVF with tubal infertility, pelvic adhesion or multiembryos transfer are risk factors of HP. Furthermore, surgery could induce abortion.

Background: Hypertension is a clinically common cardiovascular disease, resulting in many complications. Omega-3 might be beneficial in lowering blood pressure. This protocol will be performed to evaluate the effects of omega-3 on blood pressure in hypertensive patients. Methods: We will search both the electronical databases and paper-published journals. Endnote software will be used to complete study screening and data extraction by 2 reviewers independently. Review Manager software will be used to synthesize the data. The primary outcomes are systolic blood pressure and diastolic blood pressure. Secondary outcome is the adverse effects. Results: The results of this study will propose a trustworthy evidence to evaluate the effects of omega-3 on blood pressure of hypertensive patients. Conclusion: The conclusion of our systematic review will reply whether omega-3 is an effectual intervention to lower blood pressure of hypertensive patients. Ethics: This review does not require ethical approval because all of the data analyzed in this review have been published. Registration number: INPLASY202070103 (DOI number: 10.37766/inplasy2020.7.0103)

Background: Coronary artery disease (CAD) is a clinically common coronary heart disease. Vitamin D might be beneficial in CAD patients through its favorable effects on inflammation biomarkers. This study will be performed to examine the effects of Vitamin D supplementation on inflammatory markers in CAD patients. Methods: We will search the electronical databases and hand-searching journals or reference lists. The study screening and data extraction will be carried out by 2 investigators independently. The primary outcome is inflammatory biomarkers of peripheral blood. Secondary outcomes are triglyceride, total cholesterol, high-density lipoprotein cholesterol levels, low-density lipoprotein cholesterol, and blood pressure. Review Manager (RevMan, version 5.3; The Nordic Cochrane Centre, Copenhagen, Denmark: http://community.cochrane.org) V.5.3 software will be used to compute the data. Results: The results of the study will provide a reliable evidence to assess the efficacy of Vitamin D supplement on inflammation biomarkers of CAD patients. Conclusion: The conclusion of our systematic review will answer whether Vitamin D is an effective intervention to relieve inflammation of CAD patients. Ethics: Because all of the data used in this review has been published, this review does not require ethical approval. Registration number: INPLASY202060072.

OBJECTIVE: To evaluate the efficacy and safety of hydroxychloroquine (HCQ) in the treatment of patients with moderate coronavirus disease 2019 (COVID-19). METHODS: We prospectively enrolled 30 treatment-naïve patients with confirmed COVID-19 after informed consent at Shanghai Public Health Clinical Center. The patients were randomized 1:1 to HCQ group and the control group. Patients in HCQ group were given HCQ 400 mg per day for 5 days plus conventional treatments, while those in the control group were given conventional treatment only. The primary endpoint was negative conversion rate of SARS-CoV-2 nucleic acid in respiratory pharyngeal swab on days 7 after randomization. This study has been approved by the Ethics Committee of Shanghai Public Health Clinical Center and registered online (NCT04261517). RESULTS: One patient in HCQ group developed to severe during the treatment. On day 7, nucleic acid of throat swabs was negative in 13 (86.7%) cases in the HCQ group and 14 (93.3%) cases in the control group ( P>0.05). The median duration from hospitalization to virus nucleic acid negative conservation was 4 (1, 9) days in HCQ group, which is comparable to that in the control group [2 (1, 4) days, Z=1.27, P>0.05]. The median time for body temperature normalization in HCQ group was 1 (0, 2) day after hospitalization, which was also comparable to that in the control group [1 (0, 3) day]. Radiological progression was shown on CT images in 5 cases (33.3%) of the HCQ group and 7 cases (46.7%) of the control group, and all patients showed improvement in follow-up examinations. Four cases (26.7%) of the HCQ group and 3 cases (20%) of the control group had transient diarrhea and abnormal liver function ( P>0.05). CONCLUSIONS: The prognosis of COVID-19 moderate patients is good. Larger sample size study are needed to investigate the effects of HCQ in the treatment of COVID-19. Subsequent research should determine better endpoint and fully consider the feasibility of experiments such as sample size.

Introduction: Wrist stiffness is a common sequela of distal radial fractures. Manipulation is generally used and effective, but problems exist, such as intense pain, swelling during the process of manipulation and long treatment period. Therefore, a combinative therapeutic strategy is necessary to benefit rehabilitation after distal radius fracture. Shi's Yi-Qi Bu-Shen Tong-Luo decoction (BTD) combined with wrist manipulation has been used in the Shanghai Ninth People's Hospital Huangpu Branch in last few decades. BTD has potential therapeutic effects on rehabilitation after distal radius fracture, which should be evaluated by rigorous clinical trial. Methods/design: A randomized, double-blind, placebo-controlled clinical trial will be conducted to determine the efficiency of BTD in relief of wrist stiffness and pain and function rehabilitation. A total of 80 wrist stiffness patients with or without pain and edema will be enrolled, and treated with wrist manipulation plus BTD or placebo for 4 weeks. The primary outcome measure is the Cooney wrist score. The second outcome measures include pain numerical rating scale, patient rated wrist evaluation, 36-item short form health survey questionnaire, and side effects. Discussion: Although BTD has shown effects on rehabilitation after distal radius fracture in the Shanghai Ninth People's Hospital Huangpu Branch for decades, the universality of this efficacy needs evaluated. The results of this trial will provide a convincing evidence. Trial registration: ChiCTR2000029260, January 19, 2020.

Aim: Owing to the inadequate data to support the valid instrument for assessing the positive changes among patients with malignant bone tumor, the present study was designed to provide such valid evidence through examining the psychometric properties of a Chinese version of the Posttraumatic Growth Inventory (PTGI-C) among these patients, and to evaluate the effects of posttraumatic growth on positive and negative symptoms in malignant bone tumor patients. Methods: Potential patients with malignant bone tumor from five tertiary hospitals were admitted to the hospital during the period from January 2013 to October 2017. At the baseline assessment (T1), all patients completed a demographic form, PTGI-C, and Positive and Negative Affect Scale (PANAS). After 4 weeks later (T2), all the patients finished PANAS and PTGI-C again, and the PTGI-C was re-administered to patients who were simple randomly selected from the total sample. Results: The PTGI-C exhibited moderate reliability and validity. The Cronbach’s α coefficient of the total scale was 0.91. Confirmatory factor analysis supported the five-factor model and the convergent validity results obtained were distinct but correlated. The multiple linear regression analyses showed that posttraumatic growth had a significant prediction on positive affect (F=16.445, p

Rationale: Retained placenta accreta is an increasing obstetric problem in recent years, and pulmonary embolism (PE) during pregnancy and the postpartum period is a vital condition, but lack of standard therapy guidelines. This report describes a case of postpartum PE combined with retained placenta accreta. Patient concerns: A 27-year-old woman presenting with fever and dyspnea after delivery was admitted to our hospital with retained placenta accreta. Diagnoses: The patient was diagnosed with the infection, postpartum PE, and residual placenta. Interventions: The antibiotics and low molecular weight heparin were initially started to cure the infection and control PE. Mifepristone was then used to promote the necrosis of residual placenta while long-term use of warfarin was served as continuous anticoagulant therapy. Hysteroscopic resection of retained placenta was not performed until thrombi had been almost disappeared after more than 2 months of anticoagulation therapy. Outcomes: The patient's menstruation returned to normal within several weeks after hysteroscopic resection and she completely recovered from PE after 3 months of anticoagulant therapy. Lessons: Treatment of retained placenta accreta can be postponed when encountering complicated cases, such as postpartum PE. PE in perinatal stage can be managed referring to nonmaternal PE.

Rationale: Postpartum deep vein thrombosis is a unique condition in diagnosis and treatment. Rivaroxaban, a novel oral anticoagulant, is indicated for acute deep vein thrombosis, but limited data have been reported for postpartum women. Catheter-directed thrombolysis is a common procedure for treating acute deep vein thrombosis, but it is rarely used for postpartum patients, especially after more than 3 months. Patient concerns: A 31-year-old Asian woman suffered from progressive erythematous swelling and local heat of the left lower limb after twin delivery. Diagnoses: Venous duplex ultrasound examination showed thrombus formation in the left femoral vein and popliteal vein with reduced compressibility. After standard treatment of novel oral anticoagulant therapy for 4 months, we observed only partial improvement of the symptoms, and the condition deteriorated after her ordinary activities. Interventions: Venography was performed and a large amount of thrombus lining from left femoral vein to left iliac vein was noted with total occluded left common iliac vein. After catheter-directed thrombolysis and balloon dilatation, better flow was regained and her symptoms improved completely after procedure. Outcomes: During a 1-year follow-up without medication, the patient did not complain about leg swelling, exercise aggravation, or any other post-thrombotic symptoms. Lessons: Pregnancy seems to be a transient provoking factor for deep vein thrombosis, but it is sometimes refractory even during the postpartum period. Follow-up imaging studies should be encouraged to confirm the vessel condition, particularly for applying down-titration or discontinuation strategies of medication. Catheter-directed thrombolysis could be considered as an alternative method for postpartum iliofemoral deep vein thrombosis. Postpartum women usually have favorable functional status and lower bleeding risk. Rivaroxaban is a favorable choice for deep vein thrombosis, but its use in postpartum women is still controversial, and evidence of its effectiveness is not available. Thus, endovascular intervention can be a relatively safe therapy, in addition to anticoagulation therapy for premenopausal patients with recurrent deep vein thrombosis.

In this study, we propose a method for predicting the supporting pressure required for shallow tunnels in layered soils, based on a curved roof collapse mechanism with multi-failure surfaces. In this method, the effect of the number and thickness of soil layers, pore water pressure, arbitrary roof profile, and ground additional load is considered simultaneously. A nonlinear power-law failure criterion is employed to describe the failure characteristics of the roof soils. The internal energy dissipation rate and the work rates produced by external forces are obtained based on the associated flow rule and plasticity potential theory. The analytical expressions of the required supporting pressure and roof collapse surfaces are obtained with the upper bound method. Furtherly, a shallow rectangular tunnel in two soil layers is selected for parametric investigation. The change laws of the required supporting pressure and collapse curves under varying parameters are obtained. Furthermore, the corresponding engineering recommendations are given, which may potentially provide references for the support design and construction of shallow tunnels in layered strata.

Elucidating the cold tolerance mechanism of Paeonia lactiflora, which is one of the most valuable ornamental and medicinal plants in Asia, fundamentally impacts its breeding and production. The glycerol-3-phosphate acyltransferase (GPAT) gene plays a pivotal role in cold resistance in a variety of plant species. Here, we cloned the P. lactiflora GPAT gene, determined its expression pattern, and tested its role in cold resistance. We obtained the full-length P. lactiflora GPAT gene using tissue-cultured seedlings and real-time polymerase chain reaction and rapid amplification of cDNA ends analyses. We named this gene PlGPAT in P. lactiflora. Phylogenetic analysis indicates that the PlGPAT gene is closely related with the GPAT genes in core eudicots. The phylogenetic tree containing 31 angiosperm species based on GPAT protein sequences is largely consistent with the known phylogeny in flowering plants. We conducted a time-course PlGPAT expression analysis and demonstrated that PlGPAT expression is correlated with low-temperature stress. Our results suggest that the PlGPAT gene plays an important role in regulating cold resistance in P. lactiflora.

Cystoscopic technique is the current common method of retrieving double J ureteral stent in most pediatric urological centers. In this study, we evaluated the feasibility and efficacy of a novel noncystoscopic method to remove retained ureteral stents in pediatric patients. We reviewed all medical records from a total of 102 patients who were treated in our hospital between January 2013 and December 2016 to remove the double J ureteral stent retained into the ureter. The pediatric patients were divided into 2 groups based on different surgical options: cystoscopic group and noncystoscopic group. The surgery time (including time for instrument preparation), operation time, expenses, postoperative urination discomfort, and hospitalization were compared between the 2 groups. The noncystoscopic group took significantly less time for surgery and operation than the cystoscopic group (surgery time:7.40 ± 3.75 vs 18.42 ± 2.77 min, P

Purpose To investigate the potential inhibitory effects of RNA interference-mediated knockdown of neuropilin-2 (NP2) on inflammation-induced corneal hemangiogenesis and lymphangiogenesis, and whether selective inhibition of lymphangiogenesis on vascularized recipient beds before transplantation improves the graft survival. Methods Mouse lymphatic endothelial cells were transfected with the plasmid expressing artificial microRNA (amiRNA) against mouse NP2, and the down-regulation of VEGF-C-induced NP2 expression by NP2 amiRNA was evaluated by real-time PCR and western blot assays. Next, NP2 amiRNA or negative control amiRNA was injected intrastromally into BALB/c mouse model of suture-induced corneal neovascularization three days after surgery. Corneas were harvested 1 week after suture placement and the formation of lymphatic and blood vessels as well as the recruitment of macrophage was evaluated by immunohistochemical staining. The neovascularized graft beds treated by NP2 amiRNA or control then served as recipients of orthotopic corneal transplants, and age-matched C57BL/6 donors were used. Corneal allografts were examined twice a week for 8 weeks, and graft clarity was quantified by means of an opacity score. Results VEGF-C-induced NP2 expression at both mRNA and protein levels was significantly suppressed by NP2 amiRNA in mouse lymphatic endothelial cells. Intrastromal administration of NP2 amiRNA reduced corneal lymphangiogenesis by 45% versus control (p=0.015), but corneal hemangiogenesis (p=0.815) and the recruitment of CD11 antigen-like family member B (CD11b)-positive macrophage (p=0.589) were unchanged. Kaplan–Meier survival analysis revealed a better graft survival rate in the vascularized recipient beds pre-treated by NP2 amiRNA in comparison to controls (p=0.014). Conclusions Knockdown of NP2 improves corneal graft survival by selectively inhibiting lymphangiogenesis in vascularized beds before transplantation. Thus our results open new treatment options for transplant rejection and other lymphatic disorders.

Purpose Oxidative damage induced by H2O2 treatment can irreversibly damage the lens epithelium, resulting in cell death and cataract. Whether the effects of oxidative stress could be attenuated in cultured human lens epithelial cells by incubation with resveratrol (RES) is still unknown. In the present study, we examined the function of resveratrol in protecting human lens epithelial B-3 (HLEB-3) cells against H2O2 induced cell death and cell apoptosis, its role in reducing H2O2 induced intracellular reactive oxygen species (ROS) accumulation, and investigated the mechanism by which resveratrol underlies the effect. Methods HLEB-3 cells, a human lens epithelial cell line, were exposed to 100 μM H2O2 with or without RES pre-treatment at different concentrations for different time duration. Cell viabilities were monitored by 4-[3-[4-iodophenyl]-2-4(4-nitrophenyl)-2H-5-tetrazolio-1,3-benzene disulfonate] (WST-1) assay. The apoptosis rate and ROS generation were detected by flow cytometric analysis. Expression levels of superoxide dismutases-1 (SOD-1), catalase, and heme oxygenase-1 (HO-1) proteins were measured by western-blotting analysis. p38 and c-jun N terminal kinase (JNK) activation was also evaluated by western-blotting analysis. Results Resveratrol clearly reduced H2O2 induced cell apoptosis and ROS accumulation; protected HLEB-3 cells from H2O2 induced oxidative damage, and increased the expression levels of SOD-1, catalase, and HO-1. Further studies showed that RES also inhibited H2O2 induced p38 and JNK phosphorylation. Conclusions These findings suggested that RES protected HLEB-3 cells from H2O2 induced oxidative damage, presumably by inducing three antioxidative enzymes including catalase, SOD-1, and HO-1.

Purpose The objective of the present study was to investigate the efficacy of rapamycin on rabbit lens epithelial cell proliferation, migration, and secrcetion of extracellular matrix fibronectin (Fn). Methods Rabbit lens epithelium cells (rLECs) were isolated from 1 month old rabbit. rLECs were either cultured for 24, 48, or 72 h with different doses of rapamycin (0.1, 1, and 10 ng/ml). The proliferation kinetics, proliferating cell nuclear antigen (PCNA) expression, and basic fibroblast growth factor (bFGF)-induced migration of rLEC was determined by methyl thiazol tetrazolium (MTT) assay, western blotting and transwell chamber assay, respectively. The effect of rapamycin on the synthesis of Fn was examined via immunofluorescence. Results Rapamycin significantly inhibited rLEC proliferation and PCNA protein expression when administered doses and time periods except for 0.1 ng/ml for 24 h. bFGF-induced migration rLECs was inhibited by pretreatment with rapamycin for 48 h. Extracellular matrix Fn formation of rLECs was also reduced by rapamycin. Conclusions In our study, rapamycin strongly inhibited rLEC proliferation, bFGF-induced migration, and extracellular matrix Fn formation. Thus, rapamycin may have a potential inhibition of posterior capsule opacificatin (PCO) and needs further study.

Purpose To report the identification of a novel nonsense mutation in CRYGC in a Chinese family with autosomal dominant congenital nuclear cataracts and microcornea. Methods We investigated a four-generation Chinese family with six members affected with nuclear cataracts and microcornea. The family resides in a relatively isolated region of northern China. Genomic DNA was isolated from blood leucocytes, genotyping was performed using more than 100 microsatellite markers for the known cataract candidate gene loci, and LOD scores were calculated using the LINKAGE programs. Mutations were detected by DNA sequence analysis of the candidate genes. Results Evidence for linkage was detected at marker D2S325 (LOD score [Z]=2.29, recombination fraction [θ]=0.0), which closely flanks the γ-crystallin gene cluster (CRYGA-CRYGD) on chromosome 2q32.3-q35. Direct sequencing of the candidate CRYGA-CRYGD gene cluster revealed a c.470G>A transversion in exon 3 of CRYGC, which cosegregated with cataracts in the family and was not observed in 100 normal controls. This single nucleotide change was predicted to introduce a translation stop codon at tryptophan 157 (W157X). Conclusions The present study has identified a novel nonsense mutation in CRYGC associated with autosomal dominant cataracts and microcornea in a Chinese family. Our finding expands the spectrum of CRYGC mutations associated with congenital cataract and confirms the role of γ-crystallin in the pathogenesis of congenital nuclear cataracts.

Purpose To characterize the clinical phenotype, histopathological features, and molecular genetic basis of an Avellino corneal dystrophy (ACD) in a Chinese family. Methods A complete ophthalmologic examination was performed in 21 individuals (6 affected and 15 unaffected) of the four-generation family. DNA was obtained from peripheral blood leukocytes of each participant. Genetic analysis included TGFBI polymerase chain reaction (PCR) amplification and automated nucleotidic sequenceing of all 17 exons of genomic DNA. Histological analysis of corneal tissue from the proband was performed after a penetrating keratoplasty. One hundred Chinese controls were scanned for the presence of the R124H mutation by amplifying TGFBI exon 4 and then by direct sequencing of PCR products. Results The proband of the pedigree had phenotypic features consistent with diagnosis of ACD. He was homozygous for the same R124H mutation in TGFBI as previously reported in Japan and European countries. In addition, 4 affected and 7 unaffected individuals carried the same variation in the heterozygous state were identified. None of the 100 control subjects was positive for this mutation. Moreover, a variable expressivity and an apparent non-penetrance were observed in the individuals with heterozygous R124H mutation in our pedigree. After excluding the missed diagnosis or a late onset, it could be interpreted as a reduced penetrance. Conclusions We reported a novel ACD family which exhibited a reduced penetrance of phenotype in northern China. This outcome supports that although the R124H mutation is one of the genetic causes of the disease, different genetic and environmental factors may influence the expressivity and the penetrance. Uncovering the mechanism may facilitate us to inhibit the occurrence of the corneal dystrophy caused by the R124H mutation in TGFBI, irrespective of the homozygous and heterozygous mutation.

Background: Y-box binding protein 1 (YB-1) overexpression has been shown in various tumor cells including hepatocellular carcinoma (HCC); moreover, this protein can be actively secreted. Objectives: The aim of this study was to establish a method to quantify serum YB-1 and evaluate its clinical application in the clinical diagnosis of HCC. Patients and Methods: Recombinant YB-1 and two populations of its antibodies were prepared. A monoclonal antibody was specific to the N-terminus of YB-1 amino acids 134-160; and another was a polyclonal antibody. A sandwich-type chemiluminescence immunoassay (CLIA) was developed and evaluated. Levels of YB-1 and alpha fetoprotein (AFP) in serum samples from 105 HCC patients, 25 hepatitis B virus patients, 25 cirrhosis patients, and 50 healthy donors were detected using the established method and an AFP electrochemiluminescence kit. Results: The developed method was linear to 150 μg/L of YB-1 with a minimum detection limit of 0.01 μg/L. The average recoveries were between 93.9% and 109.0%. The mean intra- and inter-assay coefficients of variation (CVs) were 4.0-4.8% and 8.2-10.2%, respectively. The relationship between the concentration of diluted YB-1 and the dilution ratios gave a good linear correlation coefficient of 0.9986. The YB-1 concentration was increased in serum of HCC patients (33.0 ± 23.39 μg/L) compared to healthy individuals (13.2 ± 5.29 μg/L, P < 0.0001), patients with HBV (17.9 ± 7.49 μg/L, P = 0.0003), and patients with HBV cirrhosis (20.7 ± 8.75 μg/L, P < 0.05). Moreover, the combination of YB-1 and alpha-fetoprotein had a high sensitivity (89.5%) and reasonable specificity (62.0%) in identifying HCC. Conclusions: The established method has an acceptable performance in quantifying YB-1. In addition, serum YB-1 may aid in the diagnosis of HCC. Implication for health policy/practice/research/medical education: In our submitted manuscript, recombinant YB-1 and two populations of antibodies were prepared; and a rapid and sensitive sandwich-type chemilu- minescence immunoassay (CLIA) was developed. The concentrations of serum YB-1 in healthy donors, hepatitis B virus patients, patients with hepatic cirrhosis and HCC patients were quantitatively measured for the first time. This revealed that serum YB-1 concentrations were significantly increased in HCC compared with control groups. Furthermore, the combination of YB-1 and alpha-fetoprotein has a high sensitivity and reasonable specificity to identify HCC. Our findings suggested that this new assay is reliable for routine clinical application, and serum YB-1 levels might be a potential routine tumor marker for screening HCC, especially when tested in parallel with alpha-fetoprotein. Copyright © 2013, Kowsar Corp.; Licensee Kowsar Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http:// creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited. 1. Background

Background From field harvest to the consumer’s table, fresh citrus fruit spends a considerable amount of time in shipment and storage. During these processes, physiological disorders and pathological diseases are the main causes of fruit loss. Heat treatment (HT) has been widely used to maintain fruit quality during postharvest storage; however, limited molecular information related to this treatment is currently available at a systemic biological level. Results Mature ‘Kamei’ Satsuma mandarin (Citrus unshiu Marc.) fruits were selected for exploring the disease resistance mechanisms induced by HT during postharvest storage. Proteomic analyses based on two-dimensional gel electrophoresis (2-DE), and metabolomic research based on gas chromatography coupled to mass spectrometry (GC-MS), and liquid chromatography quadrupole time-of-flight mass spectrometry (LC-QToF-MS) were conducted. The results show resistance associated proteins were up-regulated in heat treated pericarp, such as beta-1, 3-glucanase, Class III chitinase, 17.7 kDa heat shock protein and low molecular weight heat-shock protein. Also, redox metabolism enzymes were down-regulated in heat treated pericarp, including isoflavone reductase, oxidoreductase and superoxide dismutase. Primary metabolic profiling revealed organic acids and amino acids were down-regulated in heat treated pericarp; but significant accumulation of metabolites, including tetradecanoic acid, oleic acid, ornithine, 2-keto-d-gluconic acid, succinic acid, turanose, sucrose, galactose, myo-inositol, glucose and fructose were detected. Noticeably, H2O2 content decreased, while, lignin content increased in heat treated pericarp compared to the control, which might increase fruit resistibility in response to external stress. Also, flavonoids, substances which are well-known to be effective in reducing external stress, were up-regulated in heat treated pericarp. Conclusions This study provides a broad picture of differential accumulation of proteins and metabolites in postharvest citrus fruit, and gives new insights into HT improved fruit disease resistance during subsequent storage of ‘Kamei’ Satsuma mandarin. Interpretation of the data for the proteins and metabolites revealed reactive oxygen species (ROS) and lignin play important roles in heat treatment induced fruit resistance to pathogens and physiological disorders.

The neuroprotective effects of 3,6'-disinapoyl sucrose (DISS) from Radix Polygala against glutamate-induced SH-SY5Y neuronal cells injury were evaluated in the present study. SH-SY5Y neuronal cells were pretreated with glutamate (8 mM) for 30 min followed by cotreatment with DISS for 12 h. Cell viability was determined by (3,4,5-dimethylthiazol-2-yl)-2,5-diphenylte-trazolium bromide (MTT) assay, and apoptosis was confirmed by cell morphology and flow cytometry assay, evaluated with propidium iodide dye. Treatment with DISS (0.6, 6, and 60 μmol/L) increased cell viability dose dependently, inhibited LDH release, and attenuated apoptosis. The mechanisms by which DISS protected neuron cells from glutamate-induced excitotoxicity included the downregulation of proapoptotic gene Bax and the upregulation of antiapoptotic gene Bcl-2. The present findings indicated that DISS exerts neuroprotective effects against glutamate toxicity, which might be of importance and contribute to its clinical efficacy for the treatment of neurodegenerative diseases.