Use of customized 3-dimensional printed mandibular prostheses with a dental implant pressure-reducing device in mandibular body defect: A finite element study performing multiresponse surface methodology.

Background/purpose: Segmental body defects of the mandible result in the complete loss of the affected region. In our previous study, we investigated the clinical applicability of a customized mandible prosthesis (CMP) with a pressure-reducing device (PRD) in an animal study. In this study, we further incorporated dental implants into the CMP and explored the use of dental implant PRD (iPRD) designs. Materials and methods: By employing a finite element analysis approach, we created 4 types of CMP: CMP, CMP with iPRD, CMP-PRD, and CMP-PRD with iPRD. We developed 2 parameters for the iPRD: cone length (CL) in the upper part and spring pitch (SP) in the lower part. Using the response surface methodology (RSM), we determined the most suitable structural assignment for the iPRD. Results: Our results indicate that CMP-PRD had the highest von Mises stress value for the entire assembly (1076.26 MPa). For retentive screws and abutments, CMP with iPRD had the highest von Mises stress value (319.97 and 452.78 MPa, respectively). CMP-PRD had the highest principal stress (131.66 MPa) in the anterior mandible. The iPRD reduced principal stress in both the anterior and posterior mandible. Using the RSM, we generated 25 groups for comparison to achieve the most favorable results for the iPRD and we might suggest the CL to 12 mm and the SP to 0.4 mm in the further clinical trials. Conclusion: Use of the PRD and iPRD in CMP may resolve the challenges associated with CMP, thereby promoting its usage in clinical practice.

Oral soft tissue biopsy surgery: Current principles and key tissue stabilization techniques.

There are different kinds of benign and malignant lesions in the oral cavity. Clinically, definite diagnosis can be confirmed only by doing adequate surgical biopsy and subsequent histopathological examination. Inadequate biopsy technique, unsuitable selection of the location for biopsy, inappropriate tissue handling and record of patients' information may lead to artifacts and misdiagnosis by the oral pathologists. Soft tissue stabilization is a challenge during oral surgery procedures. It needs the cooperation of operator, assistants, and patients to overcome the difficulty and ensure the successful outcome. In this article, we reviewed the procedures for clinical surgical biopsy, and raised three current tissue stabilization methods including fingers and gauze stabilization, stabilization with chalazion forceps and adapted instruments, and stabilization with retraction sutures. Moreover, some limitations were also presented. Clinician should examine the clinical characteristics of the oral lesion, the surrounding anatomical structures, and their own clinical experience and preference to select the appropriate tool. More understanding of these biopsy and tissue stabilization methods can effectively improve the biopsy procedures and obtain adequate tissues for histopathological examination and subsequent issue of an accurate pathological report.

The use of customized 3D-printed mandibular prostheses with pressure-reducing device: A clinical trial.

BACKGROUND: Segmental bone defects of the mandible result in the complete loss of the affected region. We had incorporated the pressure-reducing device (PRD) designs into the customized mandible prostheses (CMP) and conducted a clinical trial to evaluate this approach. METHODS: Seven patients were enrolled in this study. We examined the association among the history of radiotherapy, the number of CMP regions, the number of chin regions involved, and CMP exposure. RESULTS: We included five men and two women with an average age of 55 years. We excised tumors with an average weight of 147.8 g and the average weight of the CMP was 68.5 g. No significant difference between the two weights was noted (p = 0.3882). Three patients received temporary dentures and the CMP remained stable in all patients. CONCLUSION: The use of PRD in CMP may address the previous challenges associated with CMP, but further research is necessary.

The Clinical and Biological Effects of Receptor Expression-Enhancing Protein 6 in Tongue Squamous Cell Carcinoma.

There are currently no effective biomarkers for the diagnosis and treatment of tongue squamous cell carcinoma (TSCC), which causes a poor 5-year overall survival rate. Thus, it is crucial to identify more effective diagnostic/prognostic biomarkers and therapeutic targets for TSCC patients. The receptor expression-enhancing protein 6 (REEP6), a transmembrane endoplasmic reticulum resident protein, controls the expression or transport of a subset of proteins or receptors. Although it was reported that REEP6 plays a role in lung and colon cancers, its clinical impact and biological role in TSCC are still unknown. The present study aimed to identify a novel effective biomarker and therapeutic target for TSCC patients. Expression levels of REEP6 in specimens from TSCC patients were determined with immunohistochemistry. Gene knockdown was used to evaluate the effects of REEP6 in cancer malignancy (colony/tumorsphere formation, cell cycle regulation, migration, drug resistance and cancer stemness) of TSCC cells. The clinical impact of REEP6 expression and gene co-expression on prognosis were analyzed in oral cancer patients including TSCC patients from The Cancer Genome Atlas database. Tumor tissues had higher levels of REEP6 compared to normal tissues in TSCC patients. Higher REEP6 expression was related to shorter disease-free survival (DFS) in oral cancer patients with poorly differentiated tumor cells. REEP6-knocked-down TSCC cells showed diminished colony/tumorsphere formation, and they also caused G1 arrest and decreased migration, drug resistance and cancer stemness. A high co-expression of REEP6/epithelial-mesenchymal transition or cancer stemness markers also resulted in poor DFS in oral cancer patients. Thus, REEP6 is involved in the malignancy of TSCC and might serve as a potential diagnostic/prognostic biomarker and therapeutic target for TSCC patients.

MAP3K11 facilitates autophagy activity and is correlated with malignancy of oral squamous cell carcinoma.

Autophagy-related 4B (ATG4B) is a protease required for core machinery of autophagy. Phosphorylation of ATG4B promotes autophagy and is correlated with poor outcome of cancer. However, little is known about the upstream kinases for ATG4B phosphorylation and their association with clinical outcomes of cancer patients. Through siRNA library screening, MAP3K11 was identified as a potential kinase that phosphorylates ATG4B and increases its proteolytic activity. Ablation of MAP3K11 attenuated pS383/392-ATG4B protein levels and autophagic flux in oral cancer cells. Moreover, loss of MAP3K11 inhibited oral cancer cell growth, migration/invasion, and synergized starvation-reduced cell viability. MAP3K11 knock-out cancer cells also showed growth inhibition in vivo. Furthermore, the protein level of MAP3K11 was higher in tumor tissues than that in adjacent normal tissues in patients with oral squamous cell carcinoma (OSCC), comprising 179 buccal mucosa squamous cell carcinoma (BMSCC) and 249 tongue squamous cell carcinoma (TSCC). MAP3K11 protein levels were positively correlated with ATG4B and pS383/392-ATG4B levels in patients with OSCC, particularly in TSCC. In addition, high coexpression of MAP3K11 and ATG4B was associated with poor disease-specific survival in BMSCC and TSCC, while high coexpression of MAP3K11 and pS383/392-ATG4B was associated with unfavorable disease-free survival in BMSCC and TSCC. Taken together, our results indicated that MAP3K11 stimulated activity of ATG4B and autophagy, which may confer to malignancy of cancer cells. The expression of MAP3K11 and ATG4B was further associated with poor survival of OSCC, suggesting MAP3K11 could serve as a theranostic target of patients with OSCC.

The Use of Customized Three-Dimensionally Printed Mandible Prostheses with a Pressure-Reducing Device: A Finite Element Analysis in Different Chewing Positions, Biomechanical Testing, and In Vivo Animal Study Using Lanyu Pigs.

Segmental bony defects of the mandible constitute a complete loss of the regional part of the mandible. Although several types of customized three-dimension-printed mandible prostheses (CMPs) have been developed, this technique has yet to be widely used. We used CMP with a pressure-reducing device (PRD) to investigate its clinical applicability. First, we used the finite element analysis (FEA). We designed four models of CMP (P1 to P4), and the result showed that CMP with posterior PRD deployment (P4 group) had the maximum total deformation in the protrusion and right excursion positions, and in clenching and left excursion positions, posterior screws had the minimum von Mises stress. Second, the P4 CMP-PRD was produced using LaserCUSING from titanium alloy (Ti-6Al-4V). The fracture test result revealed that the maximum static pressure that could be withstood was 189 N, and a fatigue test was conducted for 5,000,000 cycles. Third, animal study was conducted on five male 4-month-old Lanyu pigs. Four animals completed the experiment. Two animals had CMP exposure in the oral cavity, but there was no significant inflammation, and one animal had a rear wing fracture. According to a CT scan, the lingual cortex of the mandible crawled along the CMP surface, and a bony front-to-back connection was noted in one animal. A histological examination indicated that CMP was significantly less reactive than control materials (p = 0.0170). Adequate PRD deployment in CMP may solve a challenge associated with CMP, thus promoting its use in clinical practice.

Effect of EGFR on SQSTM1 Expression in Malignancy and Tumor Progression of Oral Squamous Cell Carcinoma.

Oral squamous cell carcinoma (OSCC) is one of the most common types of malignant tumor. Sequestosome 1 (SQSTM1) serves as an adaptor of autophagy for degrading protein aggregates. The regulation of autophagy by EGFR and its clinical impacts are indicated in various types of cancer. However, the association of EGFR and SQSTM1 in OSCC is still unknown. Our results show that the expression levels of SQSTM1 and EGFR proteins are higher in tumor tissues than in the corresponding tumor-adjacent (CTAN) tissues of OSCC patients. The expression levels of SQSTM1 were positively associated with the EGFR expression level. High co-expression of SQSTM1 and EGFR is associated with poor prognosis in OSCC patients. Moreover, SQSTM1 expression is decreased in EGFR-knockdown cells. Cell growth and invasion/migration are also decreased in cells with single/combined knockdowns of EGFR and SQSTM1 or in SQSTM1-knockdown cells without EGFR kinase inhibitor Lapatinib treatment compared to that in scrambled cells. However, cell growth and invasion/metastasis were not significantly different between the scrambled cells and SQSTM1-knockdown cells in the presence of Lapatinib. This study is the first to indicate the biological roles and clinical significance of SQSTM1 regulation by EGFR in OSCC.

Changes in Pharyngeal Airway Space and Craniocervical Angle after Anterior Bimaxillary Subapical Osteotomy.

PURPOSE: This study explored the effects of genioplasty (Gep) and anterior subapical osteotomy of the maxilla and mandible (ASOMx+ASOMd) on the pharyngeal airway dimensions of patients with bimaxillary protrusion (BiP). METHOD: Thirty-two patients were divided into 2 groups. Group 1 received ASOMx+ASOMd, and group 2 received ASOMx+ASOMd+Gep. The cephalograms of the patients were collected before surgery and 2 months after surgery. Changes in the landmarks, related cephalometric angles (gonial, SN-GoGn, Y-axis, and SN-C2C4 angles), and 2 pharyngeal airway dimensions (uvulo-pharyngeal airway [UOP] and tongue-pharyngeal airway [TOP]) were analyzed. RESULTS: Before surgery, the parameters (incisor superius, incisor inferius, menton, most superior and anterior point of the hyoid bone, tip of the uvula, inferoanterior point on the second cervical vertebra, and inferoanterior point on the fourth cervical vertebra) and measured angles (SNA, SNB, ANB, gonial, SN-GoGn, Y-axis, and C4C2-SN) of both groups showed no significant differences. Following ASOMx, the patients in groups 1 and 2 exhibited a setback by 7.0 and 6.6 mm, respectively. After ASOMd, groups 1 and 2 exhibited 4.9 and 5.3 mm setbacks, respectively. No significant difference in the amount of setback was observed between groups 1 and 2. The postoperative horizontal and vertical positions of Me in group 2 were significantly forward by 6.1 mm and upward by 1.5 mm, respectively. Regarding pharyngeal airway dimensions, TOP was decreased in group 1 (1.7 mm) and group 2 (1.3 mm). In the postoperative Pearson correlation coefficient test, the horizontal and vertical positions of Me showed no significant correlation with TOP in both groups. Therefore, Gep did not prevent the reduction of TOP in group 2. CONCLUSION: After bimaxillary anterior subapical osteotomy, the TOP of patients with BiP was decreased, and this situation was unavoidable, regardless of whether Gep was performed.

Clinical Significance and the Role of Guanylate-Binding Protein 5 in Oral Squamous Cell Carcinoma.

Guanylate binding protein 5 (GBP5) is the interferon (IFN)-inducible subfamily of guanosine triphosphatases (GTPases) and is involved in pathogen defense. However, the role played by GBP5 in cancer development, especially in oral squamous cell carcinoma (OSCC), is still unknown. Herein, next-generation sequencing analysis showed that the gene expression levels of GBP5 were significantly higher in OSCC tissues compared with those found in corresponding tumor adjacent normal tissues (CTAN) from two pairs of OSCC patients. Higher gene expression levels of GBP5 were also found in tumor tissues of 23 buccal mucosal squamous cell carcinoma (BMSCC)/14 tongue squamous cell carcinoma (TSCC) patients and 30 oral cancer patients from The Cancer Genome Atlas (TCGA) database compared with those in CTAN tissues. Immunohistochemical results showed that protein expression levels of GBP5 were also higher in the tumor tissues of 353 OSCC patients including 117 BMSCC, 187 TSCC, and 49 lip squamous cell carcinoma patients. Moreover, TCGA database analysis indicated that high gene expression levels of GBP5 were associated with poor overall survival in oral cancer patients with moderate/poor cell differentiation, and associated with poor disease-free survival in oral cancer patients with moderate/poor cell differentiation and lymph node metastasis. Furthermore, GBP5-knockdowned cells exhibited decreased cell growth, arrest at G1 phase, and decreased invasion/migration. The gene expression of markers for epithelial-mesenchymal transition and cancer stemness was also reduced in GBP5-silenced oral cancer cells. Taken together, GBP5 might be a potential biomarker and therapeutic target for OSCC patients, especially for those with poor cell differentiation and lymph node metastasis.

Antilingula as a Surgical Reference Point for Vertical Ramus Osteotomy.

PURPOSE: This study investigated the antilingula and its related landmarks, the mandibular rami, by using cone-beam computed tomography (CBCT). METHODS: CBCT images of 37 patients (74 sides of the mandibular ramus) were collected. The landmarks of antilingula (AntiL), anterior ramus (A), posterior ramus (P), superior ramus (S), and inferior ramus (I) were identified. The distances (A-AntiL, P-AntiL, S-AntiL, and I-AntiL) were statistically evaluated according to gender, side (right and left), and skeletal patterns. RESULTS: The distance from the antilingula to the anterior (A-AntiL) border of the ramus was significantly longer on the right side (14.69 mm) than on the left side (13.97 mm). Male patients had longer AntiL-P, AntiL-I, and S-I distances (18.96, 40.07, and 54.94 mm, respectively) than did female patients (16.66, 35, and 47.54 mm, respectively). Regarding skeletal patterns, the classes can be ordered as follows in terms of the measurements: class III>class II>class I. However, the differences between the classes were nonsignificant. Pearson correlation analysis revealed that gender and S-I distance were strongly correlated (r = 0.667); specifically, male patients had a longer S-I distance. A-AntiL and A-P also exhibited a strong correlation (r = 0.796). CONCLUSION: Antilingula-related distances did not differ between skeletal patterns. Among antilingula-related variables, A-AntiL could serve as a favorable measuring point during operation.

Osteomyocutaneous Free Fibula Flap Prevents Osteoradionecrosis and Osteomyelitis in Head and Neck Cancer Reconstruction.

BACKGROUND: Osteoradionecrosis (ORN) is one of the most severe complications of free fibula reconstruction after radiotherapy. The gold standard treatment of osteomyelitis involves extensive debridement, antibiotics, and sufficiently vascularized muscle flap coverage for better circulation. Therefore, we hypothesized that free fibula flap with muscle could decrease the risk of ORN. METHODS: This study consisted of 85 patients who underwent reconstruction with free fibula flap in head and neck cancer by a single reconstructive surgeon at Kaohsiung Veterans General Hospital over a period of 19 years (1998-2016). Patients with postoperative adjuvant radiotherapy were included in the study and were grouped by either free fibula osteocutaneous flap or free fibula osteomyocutaneous flap (with flexor hallucis longus muscle), and the incidence of ORN was compared. RESULTS: Of the 85 patients, 15 were reconstructed with osteocutaneous fibula flap and 70 were with osteomyocutaneous fibula flap. The rate of ORN or osteomyelitis was significantly lower in the muscle group (18.6%, n = 13/70 vs. 46.7%, n = 7/15, p = 0.020, Chi-square test). CONCLUSION: Vascularized muscle transfer increases perfusion of surrounding tissues and the bone flap, thereby decreasing the incidence of osteomyelitis or osteonecrosis.

UBE2C is a Potential Biomarker for Tumorigenesis and Prognosis in Tongue Squamous Cell Carcinoma.

Ubiquitin-conjugating enzyme 2C (UBE2C) involves in numerous cellular processes and the tumor progression in many cancers. However, its role in oral squamous cell carcinoma (OSCC) is unclear. We aimed to investigate the role and clinical significance of UBE2C in OSCC. The expression levels of UBE2C were examined by immunohistochemistry in 185 buccal mucosa squamous cell carcinomas, 247 tongue squamous cell carcinomas (TSCCs) and 75 lip squamous cell carcinomas. The roles of UBE2C in cell growth, invasion/migration and cancer stemness were also examined in OSCC cells. The expression levels of UBE2C protein were higher in tumor tissues than they were in the corresponding tumor adjacent normal tissues from OSCC patients. Higher UBE2C expression was associated with poor cell differentiation and lymph node invasion in OSCC patients. High UBE2C expression was also correlated with shorter disease-specific survival in TSCC patients having poor cell differentiation, advanced pathological stages, lymph node metastasis as well as receiving radiation therapy. Compared to control cells, OSCC cells in which UBE2C was silenced showed decreased cell proliferation, migration/invasion and colony formation and they exhibited lower expression levels of the following cancer stemness markers-ALDH1/A2, CD44, CD166 and EpCAM. High co-expression levels of UBE2C/CD44, UBE2C/CD166 and UBE2C/EpCAM were associated with poor prognosis in oral cancer patients from The Cancer Genome Atlas database. Our findings indicated that UBE2C might be a potential biomarker for tumorigenesis and prognosis in TSCC.

Detection of the clarithromycin resistance of Helicobacter pylori in gastric mucosa by the amplification refractory mutation system combined with quantitative real-time PCR.

The goal of this study was to evaluate the feasibility of detecting Helicobacter pylori clarithromycin resistance in gastric mucosa using the amplification refractory mutation system combined with quantitative real-time PCR (ARMS-PCR). Gastric mucosal specimens (150) were collected from patients who were unsuccessfully treated for H. pylori eradication. Each specimen was divided into 2 samples. One sample was used to extract genomic DNA and detect any gene mutations of H. pylori produced by ARMS- PCR. Sequencing was used to assess the accuracy of this method. The other sample was used to culture H. pylori. The E-test minimum inhibitory concentration (MIC) was used to assess clarithromycin resistance. The results were compared with a paired chi-square test to validate the coincidence rate among the 3 methods. The coincidence rate between the sequencing and ARMS-PCR results was 98.7%, thus verifying the accuracy of ARMS-PCR. E-tests detected 144 clarithromycin resistance cases, including 45 sensitivity cases; the resistance rate was 70%. The coincidence rate between the results of the E-test and ARMS-PCR was 97.1%, and no significant difference between the 2 methods was observed. ARMS-PCR is a simple and fast method that has high sensitivity and specificity and can be used to detect the clarithromycin resistance of H. pylori in gastric mucosa. ARMS-PCR is expected to be used to study drug resistance mechanisms and use in assays of individual therapies for H. pylori eradication.

Correlation between microRNA-143 in peripheral blood mononuclear cells and disease severity in patients with psoriasis vulgaris.

This study aims to explore the correlation between microRNA-143 (miR-143) expression in peripheral blood mononuclear cells (PBMCs) and disease severity in patients with psoriasis vulgaris. From March 2014 to November 2015, 194 patients with psoriasis vulgaris (102 patients in progressive stage and 92 patients in stable stage) were selected as the case group and 175 healthy people as a control group were enrolled in this study. ELISA was used to detect the levels of IL-17 and VEGF in serum. The qRT-PCR assay was performed to detect the relative expression of miR-143 in PBMCs. Disease severity in psoriasis vulgaris patients was graded with Psoriasis Lesions Area and Severity Index (PASI). The value of miR-143 expression in PBMCs for the diagnosis of psoriasis vulgaris was evaluated using receiver operating characteristic (ROC) curve. The correlation between miR-143 expression in PBMCs and PASI scores was measured using Spearman rank correlation analysis. Compared with the control group, serum levels of IL-17 and VEGF were higher and miR-143 expression in PBMCs was lower in the case group. Furthermore, miR-143 expression in PBMCs was lower in patients in progressive stage than that in patients with stable stage. The relative expression of miR-143 in PBMCs was negatively correlated with PASI scores of patients with psoriasis vulgaris. ROC curve showed that miR-143 was a reliable and accurate biomarker of psoriasis vulgaris. Our findings suggest that miR-143 expression in PBMCs is negatively correlated the disease severity in psoriasis vulgaris.

Application of voice signal collection and analysis in traditional Chinese medicine syndrome differentiation of deficiency and excess.

OBJECTIVE: To provide more objective basis for traditional Chinese medicine (TCM) syndrome differentiation of deficiency and excess by collecting and analyzing voice signals and extracting characteristic parameters. METHODS: All of 308 samples including 150 samples of qi-deficiency, 52 yin-deficiency, 55 excess and 51 normal were collected by "Voice Collecting System of TCM" and analyzed by wavelet packet transform (WPT) combined with approximate entropy (ApEn). The characteristic parameters with remarkable differences were chosen as input vectors for support vector machine (SVM) classifier to obtain classification results. RESULTS: Comparison between the normal and non-healthy showed that ApEn values of several nodes in 2 to 3 kHz, 3.5 to 4 kHz and 5 to 8 kHz frequencies were significantly different (P≤0.05); comparison between the deficiency and excess showed that ApEn values of several nodes in 0 to 2.5 kHz and 6 to 6.5 kHz frequencies were significantly different (P≤0.05); comparison between the qi-deficiency and yin-deficiency showed that ApEn values of node in 2 to 4 kHz frequency were significantly different(P≤0.05). The outputs of SVM showed that accuracies of samples in each group had good classification results by analyzing the ApEn values in different frequencies through WPT. CONCLUSION: The methods of voice collection and analysis used in the auscultation of TCM can provide objective basis for syndrome differentiation of deficiency and excess.

The apoptotic effect of nanosilver is mediated by a ROS- and JNK-dependent mechanism involving the mitochondrial pathway in NIH3T3 cells.

Nanomaterials and nanoparticles have received considerable attention recently because of their unique properties and diverse biotechnology and life sciences applications. Nanosilver products, which have well-known antimicrobial properties, have been used extensively in a range of medical settings. Despite the widespread use of nanosilver products, relatively few studies have been undertaken to determine the biological effects of nanosilver exposure. The purpose of this study was to evaluate the toxicity of nanosilver and to elucidate possible molecular mechanisms underlying the biological effects of nanosilver. Here, we show that nanosilver is cytotoxic, inducing apoptosis in NIH3T3 fibroblast cells. Treatment with nanosilver induced the release of cytochrome c into the cytosol and translocation of Bax to mitochondria, indicating that nanosilver-mediated apoptosis is mitochondria-dependent. Nanosilver-induced apoptosis was associated with the generation of reactive oxygen species (ROS) and JNK activation, and inhibition of either ROS or JNK attenuated nanosilver-induced apoptosis. In nanosilver-resistant HCT116 cells, up-regulation of the anti-apoptotic proteins, Bcl-2 appeared to be associated with a diminished apoptotic response. Taken together, our results provide the first evidence for a molecular mechanism of nanosilver cytotoxicity, showing that nanosilver acts through ROS and JNK to induce apoptosis via the mitochondrial pathway.

[The study on the preparation and spectroscopic properties of hydroxypropyl-beta-cyclodextrin/glipizide inclusion complex].

The interaction of glipizide and hydroxypropyl-pcyclodextrin was investigated in the present paper. The stability constant and molar ratio of glipizide and hydroxypropyl-beta-cyclodextrin was calculated from the phase soluility diagram. The solid state inclusion complex of hydroxypropyl-beta-cyclodextrin/glipizide prepared by neutralization method was characterized by Fourier transform infrared spectroscopy (FTIR), differential scanning calorimetry (DSC) and powder X-ray diffraction (XRD). It was found that the phase solubility diagram of the HP-beta-CD solution showed a typical AL-type, suggesting the formation of a soluble complex of 1 : 1 molar ratio, and the stability constant was 359 L x mol(-1) at 25 degrees C. The spectra of IR, thermograph of DSC and XRD pattern of the inclusion complex were remarkably different from the glipizide and hydroxypropyl-beta-cyclodextrin/glipizide physical mixture, and indicated that hydroxypropyl-beta-cyclodextrin/glipizide inclusion complex displayed amorphous characteristic. The experiment of solubility of inclusion complex indicated that the solubility of inclusion complex increased 25-fold.

[Comparison of sustainable development status in Heilongjiang Province based on traditional ecological footprint method and emergy ecological footprint method].

By using traditional ecological footprint method and its modification, emergy ecological footprint method, the sustainable development status of Heilongjiang Province in 2005 was analyzed. The results showed that the ecological deficits of Heilongjiang Province in 2005 based on emergy and conventional ecological footprint methods were 1.919 and 0.6256 hm2 x cap(-1), respectively. The ecological footprint value based on the two methods both exceeded its carrying capacity, which indicated that the social and economic development of the study area was not sustainable. Emergy ecological footprint method was used to discuss the relationships between human's material demand and ecosystem resources supply, and more stable parameters such as emergy transformity and emergy density were introduced into emergy ecological footprint method, which overcame some of the shortcomings of conventional ecological method.

[A comparison study on the physical/chemical properties and immunomodulatory activities of mycelial polysaccharide extracts from five Ganoderma species].

Five Ganoderma species, including G. lucidum, G. tsugae, G. oerstedii, G. resinaceum and G. subamboinens, were parallel studied under an identical condition. These species were cultivated using liquid fermentation and their mycelia polysaccharides were extracted and compared on the physical/chemical properties and in vitro immunomodulatory activities. These results showed that the polysaccharide yields varied markedly among different species, and G. oerstedii was the highest among the five. However, HPLC analysis showed all the polysaccharide extracts had similar molecular weight distributions and monosaccharide compositions. They all contained glucose, galactose, mannose, glucosamine hydrochloride and fucose. In vitro assays, these polysaccharide extracts significantly stimulated phagocytosis and nitric oxide production by RAW 264.7 macrophage cell line, and G. subamboinens exerted the strongest potency. When Con A was not or presented, they all showed an up-or-down immunomodulatory effect on mouse splenocyte proliferation. The results illustrate that in addition to G. lucidum and G. tsugae, which are the two mostly studied and applied species, other Ganoderma species can also produce polysaccharides with similar physical/chemical properties and with similar immunomodulatory activities.

Facile synthetic route to enantiopure unsymmetric cis-2,5-disubstituted pyrrolidines.

The (+/-)-cis-5-arylcarbamoyl-2-ethoxycarbonylpyrrolidines 6a-g were firstly synthesized in 53-64% yields by using meso-diethyl-2,5-dibromoadipate 3 and (S)-(-)-1-phenylethylamine in three steps. The diastereomeric mixture (S;2S,5R)-(-)-7 and (S;2R,5S)-(+)-8 were prepared by the Grignard reaction and separated by a flash column chromatography in 29 and 52% yields. The absolute configurations of (+)-8 was confirmed by X-ray crystallographic analysis and the enantiopure pyrrolidines (2S,5R)-(-)-9/(2R,5S)-(+)-9 and (2S,5R)-(-)-10/(2R,5S)-(+)-10 were obtained in good yields.