A lysosomal enigma CLN5 and its significance in understanding neuronal ceroid lipofuscinosis.

Neuronal Ceroid Lipofuscinosis (NCL), also known as Batten disease, is an incurable childhood brain disease. The thirteen forms of NCL are caused by mutations in thirteen CLN genes. Mutations in one CLN gene, CLN5, cause variant late-infantile NCL, with an age of onset between 4 and 7 years. The CLN5 protein is ubiquitously expressed in the majority of tissues studied and in the brain, CLN5 shows both neuronal and glial cell expression. Mutations in CLN5 are associated with the accumulation of autofluorescent storage material in lysosomes, the recycling units of the cell, in the brain and peripheral tissues. CLN5 resides in the lysosome and its function is still elusive. Initial studies suggested CLN5 was a transmembrane protein, which was later revealed to be processed into a soluble form. Multiple glycosylation sites have been reported, which may dictate its localisation and function. CLN5 interacts with several CLN proteins, and other lysosomal proteins, making it an important candidate to understand lysosomal biology. The existing knowledge on CLN5 biology stems from studies using several model organisms, including mice, sheep, cattle, dogs, social amoeba and cell cultures. Each model organism has its advantages and limitations, making it crucial to adopt a combinatorial approach, using both human cells and model organisms, to understand CLN5 pathologies and design drug therapies. In this comprehensive review, we have summarised and critiqued existing literature on CLN5 and have discussed the missing pieces of the puzzle that need to be addressed to develop an efficient therapy for CLN5 Batten disease.

MicroRNA-93-5p/IFNAR1 axis accelerates metastasis of endometrial carcinoma by activating the STAT3 pathway.

OBJECTIVE: The aim of this study was to elucidate whether microRNA-93-5p/IFNAR1 axis promoted proliferative and metastatic changes of endometrial carcinoma (EC) cells by regulating the STAT3 signaling pathway. PATIENTS AND METHODS: The expression of microRNA-93-5p in EC tissues and cells was detected by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). Western blot was conducted to detect the protein expression of IFNAR1 in EC cells. Chi-square test was used to analyze the relationship between microRNA-93-5p expression and pathological characteristics of EC patients. The negative control, microRNA-93-5p mimics, microRNA-93-5p inhibitor or IFNAR1 vector were transfected into EC cells. Moreover, the proliferative and migratory potentials of EC cells were determined by cell counting kit-8 (CCK-8) and transwell assay, respectively. RESULTS: MicroRNA-93-5p expression was significantly upregulated in EC tissues and cells. High expression of microRNA-93-5p indicated poor survival of EC patients. The microRNA-93-5p expression level was correlated with FIGO stage and lymph node metastasis of EC. Meanwhile, a negative correlation was found between microRNA-93-5p and IFNR1 in EC tissues. Furthermore, the overexpression of microRNA-93-5p remarkably increased the viability and migratory rate of EC cells, which were reversed by IFNR1 up-regulation. CONCLUSIONS: MicroRNA-93-5p/IFNAR1 axis accelerates metastasis of EC through the STAT3 pathway.

[Effect of CIK on long-term survival in the treatment of HCC after RFA combined TACE].

OBJECTIVE: To evaluate the clinical effects of autologous cytokine-induced killer cell(CIK) on the cumulative survival and reactivation rate of hepatitics B virus(HBV) after radiofrequency ablation(RFA) combined with transcatheter arterial chemoembolization(TACE). METHODS: A total of 156 patients with hepatocellular carcinoma treated from June 2006 to September 2012 in Shengli Oilfield Central Hosptial were divided into control group(RFA, TACE) and research group(RFA, TACE, CIK). According to the tumors number, diameter and vascular invasion condition, the patients were divided into another 4 groups: the high and low risk group with tumor ≤5 cm, the high and low risk group with tumor>5 cm.The prognosis of these groups was analyzed. The effects on HBV reactivation rate between antiviral and unantiviral patients were respectively analyzed . RESULTS: The ratios of the research and control group over 1-, 3-, 5-year were 75.3%(70/93), 58.9%(53/90), 21.5%(20/93)vs 71.4%(45/63), 55.6%(35/63), 22.2%(14/63)(P>0.05), the ratios of the research and control group in the high risk group with tumor≤5 cm were 75.0%(18/24), 58.3%(14/24), 37.5%(9/24)vs 58.8%(10/17), 41.2%(7/17), 23.5%(4/17)(P<0.05). The incidences of HBV reactivation for the research and control group were 6.0% and 24.3%(P<0.05). CONCLUSION: Postoperative adjuvant CIK therapy with tumor≤5 cm after RFA combined with TACE is beneficial to the high risk group and decreases the risk of HBV reactivation.

Serum nerve growth factor level indicates therapeutic efficacy of 125I seed implantation in advanced pancreatic adenocarcinoma.

OBJECTIVE: To evaluate serum nerve growth factor (NGF) as a marker in predicting effectiveness of 125I seed implantation in advanced pancreatic carcinoma. PATIENTS AND METHODS: A total of 45 patients (30 males/15 females with mean age of 52.07±8.43 years) diagnosed with advanced pancreatic adenocarcinoma (PCa) between January 2011 to May 2014 were enrolled as PCa group in this study. Tumors were categorized as at least stage III with unresectionable condition by the TNM standard. The average tumour shortest diameter was 37.54±13.84 mm (18.50-71.20 mm). NGF level in serum before 125I seed implantation and in tumor tissue resected during surgery was measured by ELISA. After treatment, CT Scan was used to serially monitor the diameters of the tumour monthly for 6-month follow-up. RECIST was applied to evaluate the efficacy. Predictive value of serum and tumour derived NGF was evaluated based on ROC curve chart. RESULTS: We found that the serum NGF level was significantly increased in PCa patients (775.60 ± 250.97 pg/ml) compared to the healthy control group (35.03 ± 25.36 pg/ml), after age and gender adjustment. In the PCa group, the serum NGF level positively correlated with that from loci tumor tissue (r=0.487). The serum NGF level was compared between the effective group (537.42 ± 122.61 pg/ml) and noneffective group (883.17 ± 217.79 pg/ml), and significant difference was detected (p<0.0001). Patients with lower serum NGF level had good response to the 125I seeds implantation. Taking cut-off at 649.59 pg/ml, 85.70% specificity and 90.30% sensitivity were achieved by ROC. Area under the Curve of serum NGF was 0.945, standard deviation was 0.032, 95% confidence interval was 0.882-1.000. CONCLUSIONS: The level of serum NGF could be a referential index to predict the therapeutic efficacy of 125I seed implantation treatments in advanced pancreatic adenocarcinoma.

Value of alpha-fetoprotein and clinical characteristics in patients with liver neoplasm.

This article aimed to investigate the value of α-fetoprotein (AFP) for the diagnosis of hepatocellular carcinoma (HCC) and to evaluate the relationship between AFP and various clinical variables of HCC comprehensively. A retrospective study of postoperative patients diagnosed with liver neoplasm from two Chinese centers was enrolled in our study.A total of 3050 patients were included. The best cut-off point of AFP for the diagnosis of HCC was 20ng/ml with ideal sensitivity (69.74%), specificity (91.18%), LR (4.12) and YI (0.61). Non-HBV infection patients showed the highest specificity (94.44%) but lowest sensitivity (60.13%). In HBV infection. Patients, HBsAg, HBeAb, and HBcAb positive patients had the highest sensitivity (79.55%) and specificity (58.49%). AFP levels increased significantly in symptomatic patients (p=0.011). Those patients with tumor sizes ≥10cm had much higher serum AFP level compared with smaller tumors ones (p=0.014). AFP levels increased remarkably in patients with vascular invasion (p=0.015). Stepwise logistic regression showed tumor size (≥10cm) was an independent predictor of elevated AFP (OR=2.743, 95%CI: 1.167-6.447, P=0.021). The best discriminating AFP value for the diagnosis of HCC is 20ng/ml; HBsAg, HBeAb and HBcAb positive patients have the optimal sensitivity and specificity; tumor size ≥ 10cm is an independent predictor of elevated AFP.

Hepatocytes transduced with human TERT gene acquire a prolonged lifespan in culture and retain permissiveness to hepatitis B virus.

Hepatitis B virus (HBV) can be propagated in vitro in primary cultures of human hepatocytes and some stable hepatoma cell lines maintained under specific conditions. The lack of simple and non-neoplastic cell culture systems for HBV has hampered the analysis of virus life cycle and development of antiviral compounds. In this study, we succeeded in prolonging the lifespan of human hepatocytes in primary culture by transducing them with human telomerase reverse transcriptase (hTERT) gene. The transgenic cells expressed hTERT constitutively and propagated HBV up to 5x105 DNA copies/ml for 28 days.

Luteinizing hormone receptor splicing variants in bovine Leydig cells.

The luteinizing hormone receptor (LHR) plays a key role in testosterone production through its interaction with the gonadotropins, LH and chorionic gonadotropin. We examined the LHR splicing pattern in bovine Leydig cells; LH-induced expression of eight cloned splicing variants was detected by real-time PCR. Luteinizing hormone applied to cultured Leydig cells resulted in expression of full-length LHR and the A and B isoforms, as well as secretion of testosterone, which first increased, then declined, and then increased further, with increased LH levels. The secretion of testosterone progressively increased with increasing LH, but the expression levels of LHR (FL, A, and B) did not increase correspondingly. We conclude that the LHR splicing pattern is complex in bovine Leydig cells, and that expression of full-length LHR and isoforms A and B changes when induced with LH.

Reversible ferromagnetism study in un-doped ZnO thin films.

Room temperature ferromagnetism in pure ZnO thin films prepared by spin-coating method was observed. X-ray photoelectron spectroscopy and inductively coupled plasma-mass spectrometry showed no or extremely little presence of impurities, which were unlikely to be responsible for the large magnetization moment observed. In order to study the origin of ferromagnetism, ZnO thin films were rapidly annealed in N2 and O2 ambient in a repetitive way. Electrical and magnetic performance after each annealing was measured. It is found that ferromagnetism is diminished and re-appeared, in accordance with the decrease and increase of conductivity. Cathodoluminescence spectra show evidence of reversible variation of oxygen vacancy defect in the annealing process. These results provide strong evidence that oxygen vacancies play a significant role in inducing ferromagnetism in ZnO thin films.

Hepatitis C virus infection of mouse hepatoma cell expressing human CD81 or LDLR.

Hepatitis C virus (HCV) infection represents a serious public health problem worldwide. Development of new therapeutics against HCV has been hampered by the lack of a small-animal model. Until now, it has been unclear which host factors influence the HCV infection. It was known that human CD81 (hCD81) or low-density lipoprotein receptor (hLDLR), the putative HCV receptors, induce concentration of viral particles on the cell surface. In this study, recombinant plasmids containing hCD81 or hLDLR genes were transfected to mouse hepatoma Hepa 1-6 cells and transgenic cell lines expressing hCD81 (hCD81/1-6 cell line) or hLDLR (hLDLR/1-6 cell line) on their surface have been established. HCV infection of these cell lines showed that the virus was bound, entered the cell, and replicated inside the cell. This finding is essential for the development of mouse model for the study of HCV replication in vivo.

Evidence of intrinsic ferromagnetism in individual dilute magnetic semiconducting nanostructures.

Semiconductors doped with magnetic ions, also known as dilute magnetic semiconductors, are both semiconducting and ferromagnetic. It remains unclear, however, whether this ferromagnetism is intrinsic, as is required for spintronic applications, or is due instead to dopant clustering. Here, we report conclusive evidence for intrinsic ferromagnetism in individual ZnO nanoparticles doped with transition metal ions. Through a simultaneous magnetic and microstructural characterization using electron magnetic chiral dichroism and channelling-enhanced electron energy loss microanalysis, respectively, we show that ZnO nanoparticles have intrinsic ferromagnetism when doped with cobalt, but not when doped with iron.

Identification and degradation characterization of hexachlorobutadiene degrading strain Serratia marcescens HL1.

A bacterium (strain HL1) capable of growing with hexachlorobutadiene (HCBD) as sole carbon and energy sources was isolated from a mixture of soil contaminated with HCBD and activated sludge obtained from petrochemical plant wastewater treatment plant by using enrichment culture. Biochemical characteristics and phylogenetic analysis based on 16S rDNA sequence indicate that strain HL1 clearly belongs to Serratia marcescens sp. Resting cells of strain HL1 were found to remove HCBD from culture fluids with the concomitant release of chloride ion under aerobic conditions. The ranges of pH value and temperature for satisfactory growth of strain HL1 cells were from 7.0 to 8.0 and 25 to 30 degrees C, respectively. Capability of resting cells to degrade HCBD was induced by HCBD in the culture fluids. HCBD (20mg/l) was removed from culture fluids by resting cells in 4 d without lag phase, but for 50mg/l and 80mg/l HCBD 7 days were needed with lag phase. Growth of strain HL1 cells was inhibited by HCBD at the concentration up to 160mg/l. First order kinetics could be fitted to the biodegradation of HCBD by HL1 cells after lag phase at initial concentrations of 20, 50, and 80mg/l. Strain HL1 also showed strong capacity to degrade chloroprene, trichloroethylene, tetrachloroethylene, and vinyl chloride at solely initial concentration of 50mg/l. Results could offer useful information for the application of strain HL1 in bioremediation or control of HCBD-polluted environment.

Novel molecular device based on electrostatic interactions in organic polymers.

A number of researchers have reported attempts to design molecular level devices. One approach is to make use of electrostatic interactions in different parts of a polymeric molecule. This paper reports a means to achieve this by adding space charge to a molecule consisting of symmetric and asymmetric subgroups. Physically, space charge residing in a subgroup produces a dipolar charge layer thereby creating a potential trough in the polymer backbone. By lifting or lowering this potential minimum, it is possible to modify the terminal current. The effect of space charge on the potential profile in the polymer backbone was examined and the change correlated to data on carrier mobilities for OC1C10-PPV reported in the literature. Modulation of space charge in the subgroup allows the manipulation of current flow along the polymer backbone, forming the basis for the development of a molecular device. A first-order analysis suggested that such a device could have current-voltage (I-V) characteristics similar to those of a MOSFET at subthreshold, with an estimated transconductance approximately 1-2 pAV and a cutoff frequency approximately 10(15) Hz.

Variable-temperature scanning tunneling microscopy and scanning tunneling spectroscopy study on copper phthalocyanine ultrathin films on a Au(111) surface.

Variable-temperature high-resolution scanning tunneling microscopy (STM) images reveal that well-ordered copper phthalocyanine (CuPc) strips can be self-assembled by depositing CuPc molecules on a Au(111) surface. The self-assembled strips are supposed to result from the balance of the intermolecular interaction and the interaction between the molecules and substrate during annealing. The energy band (approximately 1.9-2.1 eV) of CuPc, measured by scanning tunneling spectroscopy (STS), is comparable to the optical band gap (approximately 1.7 eV). Spectroscopic measurements confirm that a dipole layer and/or an effect of image force exist at the CuPc/Au(111) interface.

A novel ethacrynic acid sensor based on a lanthanide porphyrin complex in a PVC matrix.

Lanthanide porphyrin complexes synthesized by a solid state method were used to prepare a novel ethacrynic acid (EA) sensor. The sensor, based on pentane-2,4-dionato(meso-tetraphenylporphinato)terbium [TbTPP(acac)] with an optimized membrane composition, exhibits a Nernstian response to EA- ion in the concentration range 7.4 x 10(-6)-1.0 x 10(-1) mol l-1 with a pH range from 3.2 to 6.8 and a fast response time of 30 s. The electrode shows improved selectivity towards EA- ion with respect to common co-existing ions compared with the previously reported EA sensor. As electroactive materials, lanthanide porphyrin complexes show better potentiometric response characteristics than copper porphyrin complexes. The effect of solvent mediators and lipophilic ion additives was studied and the experimental conditions were optimized. The electrode was applied to the determination of EA in human urine samples with satisfactory results.

Agrobacterium-mediated transformation of cotyledonary explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis).

A procedure for producing transgenic Chinese cabbage plants by inoculating cotyledonary explants with Agrobacterium tumefaciens strain EHA101 carrying a binary vector pIG121Hm, which contains kanamycin-resistance and hygromycin-resistance genes and the GUS reporter gene, is described. Infection was most effective (highest infection frequency) when explants were infected with Agrobacterium for 15 min and co-cultivated for 3 days in co-cultivation medium at pH 5.2 supplemented with 10 mg/l acetosyringone. Transgenic plants of all three cultivars used were obtained with frequencies of 1.6-2.7% when the explants were regenerated in shoot regeneration medium solidified with 1.6% agar. A histochemical GUS assay and PCR and Southern blot analyses confirmed that transformation had occurred. Genetic analysis of T1 progeny showed that the transgenes were inherited in a Mendelian fashion.

Medium and genotype factors influencing shoot regeneration from cotyledonary explants of Chinese cabbage (Brassica campestris L. ssp. pekinensis).

Medium conditions for reliable shoot regeneration from cotyledonary explants of Chinese cabbage were examined. Maximum shoot regeneration was obtained in the presence of 5 mg/l BA and 0.5 mg/l NAA. Shoot induction was further improved by the addition of AgNO3 as well as higher concentrations (1.2-1.6%) of agar in the regeneration medium. When 123 genotypes were tested, a large variation in regeneration frequency was observed, ranging from 95% to 0%. Shoot regeneration frequency was not related to origin and days to maturity of the genotypes. Ethylene production from cultured explants seemed to play an important role in shoot regeneration. Explants of highly responsive genotypes or if cultured on the medium solidified with a higher concentration of agar generally showed low levels of ethylene production. However, AgNO3, which also enhanced shoot induction, resulted in an increase in ethylene production. The possible interaction between ethylene and shoot regeneration is discussed.