Methionine inducing carbohydrate esterase secretion of Trichoderma harzianum enhances the accessibility of substrate glycosidic bonds.

BACKGROUND: The conversion of plant biomass into biochemicals is a promising way to alleviate energy shortage, which depends on efficient microbial saccharification and cellular metabolism. Trichoderma spp. have plentiful CAZymes systems that can utilize all-components of lignocellulose. Acetylation of polysaccharides causes nanostructure densification and hydrophobicity enhancement, which is an obstacle for glycoside hydrolases to hydrolyze glycosidic bonds. The improvement of deacetylation ability can effectively release the potential for polysaccharide degradation. RESULTS: Ammonium sulfate addition facilitated the deacetylation of xylan by inducing the up-regulation of multiple carbohydrate esterases (CE3/CE4/CE15/CE16) of Trichoderma harzianum. Mainly, the pathway of ammonium-sulfate's cellular assimilates inducing up-regulation of the deacetylase gene (Thce3) was revealed. The intracellular metabolite changes were revealed through metabonomic analysis. Whole genome bisulfite sequencing identified a novel differentially methylated region (DMR) that existed in the ThgsfR2 promoter, and the DMR was closely related to lignocellulolytic response. ThGsfR2 was identified as a negative regulatory factor of Thce3, and methylation in ThgsfR2 promoter released the expression of Thce3. The up-regulation of CEs facilitated the substrate deacetylation. CONCLUSION: Ammonium sulfate increased the polysaccharide deacetylation capacity by inducing the up-regulation of multiple carbohydrate esterases of T. harzianum, which removed the spatial barrier of the glycosidic bond and improved hydrophilicity, and ultimately increased the accessibility of glycosidic bond to glycoside hydrolases.

Artificial Cells for Cellular Behavior Mimicry Induced by Sphingomyelin Degradation.

Sphingomyelinase (SMase), a hydrolase of sphingomyelin (SM) enriched in the outer leaflet of the plasma membrane of mammalian cells, is closely associated with the onset and development of many diseases, but the specific mechanisms of SMase on the cell structure, function, and behavior are not yet fully understood due to the complexity of the cell structure. Artificial cells are minimal biological systems constructed from various molecular components designed to mimic cellular processes, behaviors, and structures, which are excellent models for studying biochemical reactions and dynamic changes in cell membranes. In this work, we presented an artificial cell model that mimics the lipid composition and content of the outer leaflet of mammalian plasma membranes for studying the effect of SMase on cell behavior. The results confirmed that the artificial cells can respond to SM degradation by producing ceramides that enrich and alter the membrane charge and permeability, thus inducing the budding and fission of the artificial cells. Thus, the artificial cells developed here provide a powerful tool to study the mechanism of action of cell membrane lipids on cell biological behavior, paving the way for further molecular mechanism studies.

Accuracy and margin quality of advanced 3D-printed monolithic zirconia crowns.

STATEMENT OF PROBLEM: Nanoparticle jetting (NPJ) is a novel ceramic 3D-printing technology with high printing accuracy. However, studies reporting the accuracy of zirconia crowns manufactured by NPJ and comparing them with conventional zirconia crowns are lacking. PURPOSE: The purpose of this in vitro study was to evaluate and compare the trueness, crown fit, and margin quality of monolithic zirconia crowns manufactured by NPJ with those milled by a computer numerical control system. MATERIAL AND METHODS: A gypsum left mandibular first molar was prepared and scanned with an intraoral scanner (TRIOS 4). Three types of monolithic crowns were manufactured through 3D printing and subtractive manufacturing (SM): NPJ (3D printing), VITA (milling), UPCERA (milling). The crowns were scanned, and the dimensional deviation (trueness) was evaluated and compared by using a software program. The triple scan method was used to measure crown fit and uniform index through precise alignment in the software program, and margin quality was also observed with an optical microscope. The data were analyzed with 1-way analysis of variance and the Tukey post hoc test (α=.05). RESULTS: The NPJ group reported better trueness of all crown and axial surfaces compared with the other SM group (P<.001), but marginal trueness (P=.601), intaglio surface (P=.596), and occlusal surface (P=.641) were statistically similar compared with the Vita milled group. All 3 groups reported clinically acceptable crown fit and uniformity with statistically similar values (P>.05). The NPJ group had more crowns judged to have flawless margin quality compared with the milled groups. CONCLUSIONS: All 3 manufacturing methods can fabricate zirconia crowns with a clinically acceptable crown fit. The NPJ system could be used to manufacture monolithic zirconia crowns with better margin quality and proximal surface trueness than milled crowns.

Influences of different source microplastics with different particle sizes and application rates on soil properties and growth of Chinese cabbage (Brassica chinensis L.).

The potentially negative effects of microplastics (MP) on agroecosystems have raised worldwide concerns. However, little is known about the negative effects of MP exposure on the soil-plant system. To fill up this knowledge gap, a pot experiment was set up, and two different MP types [high density polyethylene (HDPE) and general purpose polystyrene (GPPS)] were used, which had four particle sizes (<25, 25-48, 48-150, and 150-850 µm) at four application rates (2.5, 5, 10, and 20 g MP kg-1 soil). Some soil properties and the growth of Chinese cabbage (Brassica chinensis L.) were monitored. The results showed that (1) MP application with high application rates and relatively small particle sizes significantly enhanced the soil urease activity, which accompanied with enhanced soil pH and decreased soil available concentrations of phosphorus and potassium in some cases. (2) The exposure of MP did not significantly affect the activity of soil catalase regardless of their application rates and sizes. MP with different application rates and small sizes significantly reduced the soil sucrase activity, but the largest size of MP enhanced the activity of soil sucrase. (3) GPPS at 10-20 g kg-1 or with the sizes of <25 and 48-150 µm significantly reduced the fresh weight of Chinese cabbage, but the addition of HDPE had no remarkable effects on the fresh weight regarding of its application rates or sizes. (4) MP with high application rates and large sizes enhanced but small sizes of MP reduced the leaf soluble sugar concentration. The increasing application rates of MP and small size HDPE significantly reduced the starch concentration in the leaves of Chinese cabbage, however, the different sizes of GPPS showed limited effects on the leaf starch. The addition of MP with increasing application rates and different sizes always reduced the concentration of leaf chlorophyll. These parameters regarding to plant and soil could be used to assess the risks of MP pollution in the soil-plant systems. We found that the risks resulting from MP pollution were MP type-dependent and particle size-dependent. These findings indicate that overaccumulation of MP in the agriculture may possess an ecology risk and will negatively affect the agricultural sustainability and the food safety.

Dental Age Difference in Children with ADHD.

OBJECTIVE: The purpose of this study was to determine if changes in dental development are associated with Attention Deficit Hyperactivity Disorder (ADHD) or ADHD medications. STUDY DESIGN: This retrospective chart review evaluated the dental age of 128 patients between 6 and 16 years of age using the Demirjian method from the following two groups a) children with ADHD b) unaffected children. The ADHD group was further stratified into four groups according to the medication type. The impact of ADHD on dental age difference (the difference between dental age and chronologic age) was analyzed using T-test and the association between medication type and dental age difference was analyzed through one way ANOVA. RESULTS: The mean difference between estimated dental age and chronologic age (dental age difference) for all subjects was 0.80 years. There was no significant dental age difference in subjects with ADHD and the control group (0.78±1.28vs. 0.84 ±1.09 years respectively; P=0.75) and there was no significant difference in dental age difference and type of medication (P=0.84). CONCLUSION: No significant difference was found between children with ADHD and unaffected children with respect to dental age difference. No significant differences were found in dental age difference in the four medication groups.

Potential Involvement of MiR-30e-3p in Myocardial Injury Induced by Coronary Microembolization via Autophagy Activation.

BACKGROUND/AIMS: Coronary microembolization (CME) can lead to no-reflow or slow reflow, which is one of the important reasons for loss of clinical benefit from myocardial reperfusion therapy. MicroRNAs and autophagy are heavily implicated in the occurrence and development of almost all cardiovascular diseases. Therefore, the present study was designed to investigate the role of miR-30e-3p and autophagy in CME-induced myocardial injury rat model. METHODS: Sixty rats were randomly divided into six groups: sham, CME 1h,3h,6h,9h, and 12h (n = 10 per group). Our CME rat model was created by injecting polyethylene microspheres (42mm) into the left ventricle of the heart; the sham group was injected with same volume of normal saline. The cardiac function and serum cardiac troponin I (cTnI) level of each group was measured. HE staining and HBFP staining were used to evaluate the myocardial micro-infarction area of myocardium tissue samples. Then RT-qPCR and western blot were used to detect the expression of miR-30e-3p and, autophagy related protein LC3-II and p62, respectively. Transmission electron microscope (TEM) was used to identify autophagic vacuoles in tissue samples. RESULTS: The cardiac function of the CME 6h,9h, and 12h groups were significantly decreased compared to the sham group (P < 0.05) and the cTnI level in each group were also significantly increased (P < 0.05). The expression of miR-30e-3p in the CME 6h, 9h and 12h group were decreased significantly compared with the sham group (P < 0.05). Meanwhile, the expression of autophagy related protein LC3-II decreased significantly and p62 increased significantly in the CME 9h and 12h group (P < 0.05). TEM images showed typical autophagic vacuoles for each of the CME groups. CONCLUSIONS: Myocardial miR-30e-3p is down regulated after CME and is accompanied by inhibited autophagy and decreased cardiac function. Therefore, miR-30e-3p may be involved in CME-induced cardiac dysfunction by regulating myocardial autophagy.

Additive manufacturing of dental ceramics in prosthodontics: The status quo and the future.

PURPOSE: This review aims to summarize the available technologies, material categories, and prosthodontic applications of additive manufacturing (AM) dental ceramics, evaluate the achievable accuracy and mechanical properties in comparison with current mainstream computer-aided design/computer-aided manufacturing (CAD/CAM) subtractive manufacturing (SM) methods, and discuss future prospects and directions. STUDY SELECTION: This paper is based on the latest reviews, state-of-the-art research, and existing ISO standards on AM technologies and prosthodontic applications of dental ceramics. PubMed, Web of Science, and ScienceDirect were amongst the sources searched for narrative reviews. RESULTS: Relatively few AM technologies are available and their applications are limited to crowns and fixed partial dentures. Although the accuracy and strength of AM dental ceramics are comparable to those of SM, they have the limitations of relatively inferior curved surface accuracy and low strength reliability. Furthermore, functionally graded additive manufacturing (FGAM), a potential direction for AM, enables the realization of biomimetic structures, such as natural teeth; however, specific studies are currently lacking. CONCLUSIONS: AM dental ceramics are not sufficiently developed for large-scale clinical applications. However, with additional research, it may be possible for AM to replace SM as the mainstream manufacturing technology for ceramic restorations.

Clinical efficacy of a two-piece abutment conforming to the concept of one abutment one time in the posterior region: A clinical pilot study.

OBJECTIVES: To assess the impact of a two-piece abutment workflow on enhancing the stability of the alveolar bone and gingiva surrounding the dental implant, and to determine the level of patient satisfaction. MATERIALS AND METHODS: A total of 48 patients with dentition defect in the posterior region were included and divided into two groups: the two-piece abutment workflow (TAW) and the sealing screw with submerged healing workflow (SHW). Marginal bone level (MBL), soft tissue indicators, oral hygiene indicators, and patient satisfaction were assessed and recorded partially at 0, 3, 6, and 12 months after surgery. The primary outcome was the change of MBL in different time periods. A generalized linear mixed model (GLMM) was used to take into account the correlated nature of the data, and adjust for potential confounding factors within inter-group differences. RESULTS: The survival rate of implants and prosthesis reached 100% at 12-month follow-up, with an average decrease of 0.25 mm (SD 0.23 mm) of MBL in the TAW group and 0.48 mm (SD 0.45 mm) in the SHW group. The change of MBL in the TAW group (0.15 ± 0.31 mm) was significantly lower than the SHW group (0.41 ± 0.41 mm) through the analysis of GLMM within 6 months, while no significance was found in 12 months. Moreover, less gingival pain and oppression during prosthesis loading, and less time consumption overall duration were showed in the TAW group through Visual Analogue Scale (VAS, p < 0.05). CONCLUSIONS: Within a 6-month period, the two-piece abutment workflow showed superior efficacy in preserving the integrity of the marginal bone level. Furthermore, it streamlined treatment procedures and mitigated discomfort, hence increasing patient satisfaction.

Development and evaluation of an inactivated coxsackievirus A16 vaccine in gerbils.

Coxsackievirus A16 (CVA16) is one of the major pathogens responsible for human hand, foot, and mouth disease (HFMD), which has threatened the health of young children, particularly in Asia-Pacific nations. Vaccination is an effective strategy for protecting children from CVA16 infection. However, there is currently no licensed CVA16 vaccine for use in humans. In this study, we isolated a high-growth CVA16 virus strain in MRC-5 cells and developed an MRC-5-adapted vaccine candidate strain termed CVA16-393 via two rounds of plaque purification. The CVA16-393 strain was grouped into the B1b subgenotype and grew to a titre of over 107 TCID50/ml in MRC-5 cells. The VP1 gene region of this strain, which contains the major neutralizing epitopes, displayed high stability during serial passages. The inactivated whole-virus vaccine produced by the CVA16-393 strain induced an effective neutralizing antibody response in Meriones unguiculatus (gerbils) after two doses of intraperitoneal inoculation. One week after the booster immunization, the geometric mean titres of the neutralizing antibodies for the 10246, 40812TXT, 11203SD, TJ-224 and CA16-194 strains from different regions of China were 137.8, 97.8, 113.4, 64.1 and 122.3, respectively. A CVA16 vaccine dose above 25 U was also able to provide 100% cross-protection against lethal challenges with these five clinical strains in gerbils. Immunization at a one-week interval could maintain a high level of neutralizing antibody titres for at least 8 weeks. Thus, the vaccine produced by this CVA16-393 strain might be promising.

Double-barrel fibula vascularized free flap with dental rehabilitation for mandibular reconstruction.

PURPOSE: The vascularized free fibula flap has become the most popular reconstruction method after mandibular resection because of adequate bone graft length and acceptance of dental implants. However, using 1 fibula bone may produce a height discrepancy between the native mandible and the grafted fibula that results in subsequent difficulty in wearing conventional dentures or osteointegrated implants. Several options can be used to resolve this problem such as delayed onlay bone graft, iliac bone reconstruction, fibula distraction, and double-barrel fibula flap graft. This article describes the reconstruction of segmental mandible defects with the double-barrel fibula flap and denture rehabilitation. MATERIALS AND METHODS: This procedure was used in 7 patients. A donor site fibula corresponding to at least twice the length of the mandibular defect was harvested. Double-barrel free fibula grafting was performed in 6 patients for primary reconstruction and 1 patient for secondary reconstruction, including 3 cases of osteocutaneous flap with skin islands in the reconstruction of a composite defect from a malignant tumor. Prosthodontic treatment was completed in all 7 patients. Four patients received secondary implant-supported dental reconstruction, and 3 patients who received radiation (6,000 cGy) after graft surgery had conventional removable partial dentures. RESULTS: Microvascular fibula transfers were completely successful, and all skin paddles survived without necrosis. The original mandibular contour was maintained by a reconstruction plate; the reconstruction mandibular length was 6.5 to 10 cm, the reconstruction height of the double-barrel fibula was 3.0 to 3.8 cm, and all patients were satisfied with the postoperative facial esthetics and chewing function from the implant-supported denture and removable partial prostheses. CONCLUSIONS: Mandibular segmental defects can be esthetically and functionally reconstructed by a double-barrel vascularized fibula flap that not only matches the height of the native mandible but also allows osteointegrated dental implantation for dental rehabilitation.

Clinical management of peripheral ameloblastoma.

Peripheral ameloblastoma is a rare epithelial odontogenic tumor, limited to the soft tissues of the gingiva or oral mucosa. Peripheral ameloblastoma represents approximately 2% to 10% of all ameloblastomas. It is always considered to be benign, but occasionally it may be locally aggressive or with malignant potential. In this article, we report 3 new cases of benign peripheral ameloblastoma and further discuss the clinical management of this disease.

Design and development of a new type of phimosis dilatation retractor for children.

BACKGROUND: Phimosis is one of the most common diseases in children. Early selection of appropriate treatment for phimosis in children is beneficial to the development of their reproductive organs and significantly improves the prognosis of phimosis in children. Although traditional circumcision is the most widely used, it has many disadvantages, including postoperative bleeding and incision infection, pain, obvious scars on the surgical incision, and unsatisfactory appearance. In addition, there is much controversy regarding treatment options and timing at home and abroad. Surgical procedures such as circumcision and cerclage for children with excessively long foreskin will greatly affect the normal life of children after the operation. Young children need general anesthesia, but this anesthesia carries a great risk. AIM: To design a new children phimosis dilatation retractor for children phimosis. METHODS: The children phimosis was dilated with an elastic dilation frame, in order to expand the foreskin mouth and expose the penis head, and after that, the phimosis was cured. RESULTS: A new type of phimosis dilatation retractor was designed, which can gently dilate the prepuce at multiple angles and in multiple directions at the same time. It has obtained the national patent for clinical application. CONCLUSION: The phimosis dilatation retractor based on the principle of elastically expanding the prepuce can achieve the purpose of expanding the phimosis. The clinical application shows that the effect of the children phimosis retractor is significant, which is worth promoting.

Free fibula osteocutaneous flap for primary reconstruction of T3-T4 gingival carcinoma.

Lower gingival squamous cell carcinoma (SCC) frequently invades the mandibular bone and buccal or lingual oral mucosa. In the concept of en bloc surgery of malignant tumors, it is advisable to prefer segmental mandibulectomy for T3-T4 lower gingival carcinoma that had radiologic bone involvement and resection of soft tissue on the buccal or lingual side with negative border of margin. Consequential defects of oral mucosa and mandible need immediate reconstruction to provide the maximum probability of cure and quality of life with minimal donor site morbidity. The aim of this study was to evaluate the fibula osteocutaneous flap with skin island as a means to meet both hard and soft tissue reconstructions needed in a one-stage procedure of gingival SCC. Data of 17 patients, with gingival SCC pathologically and who underwent en bloc operations including segmental mandibulectomy and reconstruction of mandible and intraoral mucosa with fibular flap, were retrospectively analyzed. The segmental mandibular defects ranged from 8 to 17 cm, and intraoral soft tissue defects ranged from 4 degrees at 2.5 cm to 8.5 degrees at 4 cm. The flaps survived in all 17 patients including 9 patients who received postoperative radiotherapy with good final function of the lower leg. Of 17 patients, 11 had with no evidence of disease with a mean follow-up period of 25 months. Our study results, together with literature findings, revealed that the fibula that had a long length of good-quality bone and sufficient blood supply were suitable for stable osteosynthesis, with the overlying skin suitable in thickness and without limitation of skin flap size for intraoral reconstruction especially after ablative surgery. This method provides oral and maxillofacial surgeons with a means to meet both hard and soft tissue needs in a one-stage procedure for extensive resection of gingival SCC.

The functioning of a novel protein, Swollenin, in promoting the lignocellulose degradation capacity of Trichoderma guizhouense NJAU4742 from a proteomic perspective.

The functioning of a novel auxiliary enzyme, TgSWO from Trichoderma guizhouense NJAU4742, was investigated based on the proteomic analysis of wild-type (WT), knockout (KO) and overexpression (OE) treatments. The results showed that the cellulase and hemicellulase activities of OE and WT were significantly higher than those of KO. Simultaneously, tandem mass tag (TMT) analysis results indicated that cellulases and hemicellulases were significantly upregulated in OE, especially hydrophobin (HFB, A1A105805.1) and endo-ß-1,4-glucanases (A1A101831.1), with ratios of 43.73 and 9.88, respectively, compared with WT. The synergistic effect of TgSWO on cellulases increased the reducing sugar content by 1.45 times in KO + TgSWO (1.8 mg) compared with KO, and there was no significant difference between KO + TgSWO (1.2 mg) and WT. This study elucidated the function of TgSWO in promoting the lignocellulose degradation capacity of NAJU4742, which provides new insights into the efficient conversion of lignocellulose.

Multifunctional DNA Polymer-Assisted Upconversion Therapeutic Nanoplatform for Enhanced Photodynamic Therapy.

Although considerable clinical attempts on various kinds of cancers have been made, photodynamic therapy (PDT) still suffers from attenuated therapeutic effects because of the developed resistance of cancer cells. As a novel antiapoptosis protein, survivin has been demonstrated to be selectively overexpressed in a great number of human malignancies and plays a significant part in cancer progression and therapeutic resistance. Herein, we present an upconversion nanoplatform for enhanced PDT by DNAzyme-mediated gene silencing of survivin. In our system, a long single-stranded DNA (ssDNA) with a repetitive aptamer (AS1411) and survivin-targeted DNAzyme was fabricated by rolling circle amplification (RCA) and adsorbed on the upconversion nanoparticles (UCNPs) by electrostatic attraction. The multivalence of the ssDNA endows the upconversion nanoplatform with high recognition and loading capacity of photosensitizers and DNAzymes. When the nanoplatform is targeted internalized into cancer cells, PDT can be triggered by near-infrared (NIR) light to generate reactive oxygen species (ROS) for killing the cancer cells. Moreover, the encoded DNAzyme can efficiently inhibit the gene expression of survivin, providing the potential to enhance the efficiency of PDT. This study thus highlights the promise of an upconversion photodynamic nanoplatform for admirable combination therapy in cancer.

Intralesional injection of Pingyangmycin may be an effective treatment for epulis.

Gingival epulis represents a family of benign tumors and tumorlike masses of the gingiva. A spectrum of inflammatory and other reactive changes can be observed pathologically as granulation, fibrous or vascular tissue. It may occur at any age, but most commonly found in patients at their twenties and sixties. Women are more frequently affected than men. Surgical excision or removal is the treatment of choice, and wider resection including extraction of the involved teeth and adjacent tissues is often required in order to prevent recurrence. However, wider resection may result in cosmetic and functional problems. Epulis bears some similarities with hemangioma both clinically and histopathologically. Therefore, we hypothesize that intralesional injection of Pingyangmycin may be a safe and effective treatment for epulis. The efficacy of this treatment modality is worthy of further investigation.

Global Gene Expression Analysis of the Brainstem in EV71- and CVA16-Infected Gerbils.

Enterovirus 71 (EV71) and coxsackievirus A16 (CVA16) are the two most important pathogens of hand, foot, and mouth disease (HFMD). However, the neuropathogenesis of EV71 and CVA16 has not been elucidated. In our previous study, we established gerbils as a useful model for both EV71 and CVA16 infection. In this work, we used RNA-seq technology to analyze the global gene expression of the brainstem of EV71- and CVA16-infected gerbils. We found that 3434 genes were upregulated while 916 genes were downregulated in EV71-infected gerbils. In CVA16-infected gerbils, 1039 genes were upregulated, and 299 genes were downregulated. We also found significant dysregulation of cytokines, such as IP-10 and CXCL9, in the brainstem of gerbils. The expression levels of 10 of the most upregulated genes were confirmed by real-time RT-PCR, and the upregulated tendency of most genes was in accordance with the differential gene expression (DGE) results. Our work provided global gene expression analysis of virus-infected gerbils and laid a solid foundation for elucidating the neuropathogenesis mechanisms of EV71 and CVA16.

A porous photocurable elastomer for cell encapsulation and culture.

Encapsulating cells within a polymer matrix creates a three-dimensional (3D) scaffold that may more accurately represent the native microenvironment and cell organization. Here we report a porous scaffold prepared from a photocurable elastomer, poly(glycerolco-sebacate)-acrylate (PGSA). The scaffold porosity, swelling, mass loss, toxicity and mechanical properties, suggest that porous PGSA could be used to support the growth and differentiation of encapsulated cells. Neuroblastoma (NB) and human embryonic stem cells (hESCs) were encapsulated into the matrix and found to adhere to the material and interact with each other within 24h. After 7 days, encapsulated NB cells were found to grow, and form matrix fibrils and tissue. Undifferentiated hESCs proliferated and differentiated in the PGSA scaffold. In vivo experiments showed that both porous scaffolds have similar biocompatibility profiles as non-porous PGSA, but porous PGSA promotes tissue ingrowth, as compared to non-porous PGSA. We therefore propose that porous PGSA scaffolds can provide a logistical template for 3D growth of cells and tissue engineering.

Expression of E-cadherin in cervical lymph nodes from primary oral squamous cell carcinoma patients.

BACKGROUND: Oral squamous cell carcinoma (OSCC) is the most common malignant tumor in oral and maxillofacial region with poor prognosis. E-cadherin plays a key role in cell-to-cell adhesion. E-cadherin expression in the metastatic cervical lymph node, especially in the micrometastatic cervical lymph node has seldom been reported in OSCC patients. OBJECTIVE: To investigate the E-cadherin expression in cervical lymph nodes from OSCC patients as well as its clinical significance. DESIGN: Thirty-three OSCC patients were involved in this study; among them, there were 28 males and 5 females, the age ranged from 34 to 78 years (mean 58.8 years). The most suspicious metastatic cervical lymph node (total 99 lymph nodes) from three cervical regions of each OSCC patient was selected for detection of E-cadherin using routine pathological examination and immunohistochemistry. RESULTS: Increased E-cadherin expression in the metastatic cervical lymph nodes was detected, which was diagnosed by routine pathological examination using HE staining. However, in the micrometastatic cervical lymph node, E-cadherin expression was negative. The survival rate of OSCC patients correlated with decreased E-cadherin expression (P=0.001), N stage (P=0.024) and tumor recurrence (P<0.001). Tumor recurrence is the only independent factor on the prognosis (RR=20.83 and P=0.014). CONCLUSIONS: Decreased E-cadherin expression in cancerous tissue correlates with the poor prognosis of OSCC patients. Detection of E-cadherin expression is useful to confirm the cervical lymph node metastasis and maybe useless to detect the cervical lymph node micrometastasis; further studies are encouraged to reveal the detail mechanism of E-cadherin expression in formation of lymph node metastatic focus.

Coxsackievirus A16 induced neurological disorders in young gerbils which could serve as a new animal model for vaccine evaluation.

Coxsackievirus A16 (CA16) is one of the major pathogens associated with human hand, foot, and mouth disease (HFMD) in the Asia-pacific region. Although CA16 infections are generally mild, severe neurological manifestations or even death has been reported. Studies on CA16 pathogenesis and vaccine development are severely hampered because the small animal models that are currently available show major limitations. In this study, gerbils (Meriones unguiculatus) were investigated for their suitability as an animal model to study CA16 pathogenesis and vaccine development. Our results showed that gerbils up to the age of 21 days were fully susceptible to CA16 and all died within five days post-infection. CA16 showed a tropism towards the skeletal muscle, spinal cord and brainstem of gerbils, and severe lesions, including necrosis, were observed. In addition, an inactivated CA16 whole-virus vaccine administrated to gerbils was able to provide full protection to the gerbils against lethal doses of CA16 strains. These results demonstrate that gerbils are a suitable animal model to study CA16 infection and vaccine development.