Oral and oropharyngeal NUT carcinoma: a multicentre screening study of poorly differentiated oral cancer.

BACKGROUND AND AIMS: Nuclear protein testis (NUT) carcinoma (NC) is a rare and highly aggressive tumour characterised by chromosomal rearrangement of the nuclear protein testis family member 1 (NUTM1) gene, also known as the NUT gene. NC occurs mainly in the head and neck, mediastinum and lung. In general, primary NC in the oral cavity is extremely rare and reported sporadically. METHODS: A total of 111 formalin-fixed and paraffin-embedded specimens of poorly differentiated oral and oropharyngeal tumours were collected from 10 hospitals. NUT protein IHC staining was performed on these samples, and fluorescence in-situ hybridisation (FISH) and RNA sequencing detection were further carried out for NUT IHC-positive cases. RESULTS: The expression of NUT protein in tumour cells was detected in five cases (five of 111, 4.5%). The tumours in these cases were located in the oral floor, lip, base of the tongue, gingiva and hard palate. FISH detection results showed BRD4::NUT rearrangement in three patients and a non-BRD4::NUT rearrangement pattern in two patients. RNA sequencing results confirmed BRD4::NUT rearrangement in two cases. CONCLUSIONS: To our knowledge, this is the first and largest retrospective study of oral NC, and we found that NC is easily misdiagnosed as poorly differentiated oral squamous cell carcinoma (SCC) or poorly differentiated carcinoma. The morphology and immunophenotype of four NC cases were similar to SCC, and abrupt keratinisation was observed in three cases. Therefore, it is necessary to detect NUT protein for NC screening in oral malignant tumours with these morphologies, especially for young patients who are more likely to be misdiagnosed with other types of cancer.

Repeated CT-guided percutaneous radiofrequency thermocoagulation for recurrent trigeminal neuralgia.

AIMS: To investigate the long-term outcomes of repeated percutaneous radiofrequency thermocoagulation (PRT) for recurrent trigeminal neuralgia (TN) patients. METHODS: Between 2002 and 2012, 33 patients with recurrent TN following an initial PRT procedure were retrospectively studied and underwent 43 repeated PRT procedures. RESULTS: The mean length of follow-up after repeated PRT was 34 months. Pain relief was immediate in 30 patients (90.9%), and no pain relief occurred in 3 patients (9.1%) following a second PRT procedure. The percentage of patients who remained in an 'excellent' and 'good' pain relief condition (pain intensity ≤BIN grade III) after the second PRT procedure was 75% at 1 year, 68% at 2 years and 68% at 5 years, and 22 of these patients (54.5%) remained satisfied with their pain relief during the follow-up period. Nine patients underwent PRT three times and 1 patient four times. The total number of patients who benefited from repeated PRT was 28 (84.8%). Postprocedure complications including masseter weakness were present in 3 patients and limited mouth opening affected 1 patient. No mortalities were observed during or after repeated PRT procedures. CONCLUSION: Repeated PRT provides long-term pain relief benefits to patients with recurrent TN and should be considered as an alternative treatment for recurrent TN.

Long-term outcome of computed tomography-guided percutaneous radiofrequency thermocoagulation for classic trigeminal neuralgia patients older than 70 years.

The incidence of trigeminal neuralgia (TN) in elderly patients is higher. However, for those with poor fitness, the optimal surgical treatment for those refractory to medical treatment is controversial. The aim of current study was to investigate the long-term outcome of computed tomography (CT)-guided percutaneous radiofrequency thermocoagulation (PRT) for 304 TN patients 70 years or older. We conducted a retrospective study of 304 elderly patients with TN who were treated with CT-guided PRT between 2002 and 2012. Follow-up was censored at the time of last contact, additional surgery, or death. Sixty-seven patients (22.1%) were of more than American Society of Anesthesiologists classification system physical status II. Excellent pain relief was 100% at discharge, 85% at 1 year, 75% at 3 years, 71% at 5 years, and 49% at 10 years. Pain relief outcomes were correlated with facial numbness. Lower temperature group (≤75°C) can attain the same long-term pain relief as higher temperature group (≥80°C); however, the incidence of painful dysesthesia rate of higher temperature group was higher than lower temperature group. Postoperative morbidity included facial numbness, masseter weakness, corneitis, hearing loss, dropping eyelid, and limited mouth opening. There were no mortalities observed during or after PRT. Our result showed CT-guided PRT is safe and effective for classic TN patients 70 years or older, including poor-fitness patients (American Society of Anesthesiologists classification system physical status >II). Lower temperature (≤75°C) is recommended for PRT in the treatment of TN.

Biodegradable ferroelectric molecular crystal with large piezoelectric response.

Transient implantable piezoelectric materials are desirable for biosensing, drug delivery, tissue regeneration, and antimicrobial and tumor therapy. For use in the human body, they must show flexibility, biocompatibility, and biodegradability. These requirements are challenging for conventional inorganic piezoelectric oxides and piezoelectric polymers. We discovered high piezoelectricity in a molecular crystal HOCH2(CF2)3CH2OH [2,2,3,3,4,4-hexafluoropentane-1,5-diol (HFPD)] with a large piezoelectric coefficient d33 of ~138 picocoulombs per newton and piezoelectric voltage constant g33 of ~2450 × 10-3 volt-meters per newton under no poling conditions, which also exhibits good biocompatibility toward biological cells and desirable biodegradation and biosafety in physiological environments. HFPD can be composite with polyvinyl alcohol to form flexible piezoelectric films with a d33 of 34.3 picocoulombs per newton. Our material demonstrates the ability for molecular crystals to have attractive piezoelectric properties and should be of interest for applications in transient implantable electromechanical devices.

Adhesive interaction of functionalized particles and endothelium in idealized microvascular networks.

OBJECTIVE: Leukocytes play a key role in the early response to tissue injury/infection resulting from physical, chemical or biological stimuli. This process involves the initiation of the leukocyte adhesion cascade mediated by a series of interactions between receptors and ligands on the endothelium and the leukocytes. Here, we characterize the adhesion profile of functionalized particles under physiological flow conditions in an idealized synthetic microvascular network (SMN) characterized by a bifurcation. We hypothesize that differences in the level of adhesion of functionalized particles in bifurcating SMNs are dependent on the ratio of adhesion molecules on the particles as well as geometric features of the in vitro networks. METHODS: Functionalized particles were prepared by coating their surfaces with different ratios of antibodies against ICAM-1 and E-selectin (aICAM-1:aE-selectin=100:0, 70:30, 50:50, 30:70, and 0:100). The adhesion of functionalized particles to 4h TNF-α activated human umbilical vein endothelial cells under shear flow (0.5, 2, and 4dyn/cm(2)) in bifurcating SMNs and in a parallel plate flow chamber was then quantified. RESULTS: The level of adhesion of 50:50 aICAM-1:aE-selectin particles was significantly higher compared to other particles in the bifurcating SMNs (~1.5-4 fold higher). However, in the parallel plate flow chamber 70:30 aICAM-1:aE-selectin particles exhibited a significantly higher level of adhesion (~1.5-2.5 fold higher). Furthermore, the adhesion of particles in junction regions was about 3-18 fold higher than that in straight sections of the SMNs. As expected, in straight sections of the SMNs and in the parallel plate flow chamber particle adhesion increased with decreasing shear. However, particle adhesion did not change significantly with decreasing shear at the junction regions of SMNs for all functionalized particles. CONCLUSION: Adhesion efficiency of functionalized particles is significantly affected by cell-adhesion molecule ratio density as well as geometric features of the vessels. Moreover, the differential adhesion patterns of particles between straight sections of bifurcating SMNs and parallel plate flow chamber, as well as straight sections and junction regions of bifurcating SMNs, indicates that adhesion profile of particles is highly dependent on the vascular geometry of the system used.

The efficacy and safety of the Xuesaitong soft capsule in the treatment of patients with ischemic stroke: systematic review and meta-analysis.

BACKGROUND: Ischaemic stroke is a common neurological disease and a leading cause of severe disability and death in developed countries. In most cases, stroke is thought to be a multifactorial disorder or complex trait for which classic patterns of inheritance cannot be shown. Xuesaitong is one of the most commonly used medicines for treating ischemic stroke in China. However, compared to the conventional therapy, the effectiveness and safety of Xuesaitong for ischemic stroke needs to be further systematically reviewed and determined. METHODS: Relevant randomized controlled trials (RCTs) examining the use of the Xuesaitong soft capsule in the treatment of patients with ischemic stroke were identified from databases, including the China National Knowledge Infrastructure, Wanfang, PubMed, Embase, and Web of Science databases. Next, 2 researchers independently extracted information from the included studies, analyzed the data using STATA 15.0 software, and evaluated the quality of the included studies using RevMan 5.3. RESULTS: A total of 17 RCTs (comprising 1,942 patients with ischemic stroke) were included in the meta-analysis. The meta-analysis results showed that the Xuesaitong soft capsule treatment increased patients' total effective rate compared to conventional or other drug treatments, and improved patients' Clinical Severity Score (CSS scores) or Barthel index (BI) score. A further subgroup analysis stratified by different treatment times showed that Xuesaitong soft capsule treatment at 4 and 8 weeks improved CSS scores more than treatment at 2 weeks in patients with ischemic stroke. Additionally, the Xuesaitong soft capsule also significantly improved plasma viscosity, whole-blood viscosity at high and low shear rates, fibrinogen, hematocrit, and the effect on traditional Chinese medicine (TCM) single symptoms or signs in patients with ischemic stroke. DISCUSSION: In summary, compared to conventional or other drug treatments, the Xuesaitong soft capsule treatment was beneficial in improving patients' TCM symptoms (e.g., crooked mouth and tongue, and dizziness) and various indicators. Further, Xuesaitong soft capsule may be a safe and effective drug for the treatment of ischemic stroke. And large-scale randomized clinical trials are needed to further confirm our findings.

One-Step Reverse-Transcription Recombinase Polymerase Amplification Using Lateral Flow Strips for the Detection of Coxsackievirus A6.

Hand, foot, and mouth disease (HFMD) is a common infectious disease affecting mainly children under 5 years of age. Coxsackievirus A6 (CVA-6), a major causative pathogen of HFMD, has caused outbreaks in recent years. Currently, no effective vaccine or antiviral treatments are available. In this study, one-step reverse-transcription recombinase polymerase amplification (RT-RPA), combined with a disposable lateral flow strip (LFS) assay, was developed to detect CVA-6. This assay can be performed in less than 35 min at 37°C without expensive instruments, and the result can be observed directly with the naked eye. The sensitivity of the RT-RPA-LFS was 10 copies per reaction, which was comparable to that of the conventional real-time quantitative polymerase chain reaction (qPCR) assays. Moreover, the assay specificity was 100%. The clinical performance of the RT-RPA-LFS assay was evaluated using 142 clinical samples, and the coincidence rate between RT-RPA-LFS and qPCR was 100%. Therefore, our RT-RPA-LFS assay provides a simple and rapid approach for point-of-care CVA-6 diagnosis.

Simultaneous delivery of chemotherapeutic and thermal-optical agents to cancer cells by a polymeric (PLGA) nanocarrier: an in vitro study.

PURPOSE: To test the effectiveness of a dual-agent-loaded PLGA nanoparticulate drug delivery system containing doxorubicin (DOX) and indocyanine green (ICG) in a DOX-sensitive cell line and two resistant cell lines that have different resistance mechanisms. METHODS: The DOX-sensitive MES-SA uterine sarcoma cell line was used as a negative control. The two resistant cell lines were uterine sarcoma MES-SA/Dx5, which overexpresses the multidrug resistance exporter P-glycoprotein, and ovarian carcinoma SKOV-3, which is less sensitive to doxorubicin due to a p53 gene mutation. The cellular uptake, subcellular localization and cytotoxicity of the two agents when delivered via nanoparticles (NPs) were compared to their free-form administration. RESULTS: The cellular uptake and cytotoxicity of DOX delivered by NPs were comparable to the free form in MES-SA and SKOV-3, but much higher in MES-SA/Dx5, indicating the capability of the NPs to overcome P-glycoprotein resistance mechanisms. NP-encapsulated ICG showed slightly different subcellular localization, but similar fluorescence intensity when compared to free ICG, and retained the ability to generate heat for hyperthermia delivery. CONCLUSION: The dual-agent-loaded system allowed for the simultaneous delivery of hyperthermia and chemotherapy, and this combinational treatment greatly improved cytotoxicity in MES-SA/Dx5 cells and to a lesser extent in SKOV-3 cells.

Liposomal Antitumor Vaccines Targeting Mucin 1 Elicit a Lipid-Dependent Immunodominant Response.

The tumor-associated antigen mucin 1 (MUC1) has been pursued as an attractive target for cancer immunotherapy, but the poor immunogenicity of the endogenous antigen hinders the development of vaccines capable of inducing effective anti-MUC1 immunodominant responses. Herein, we prepared synthetic anti-MUC1 vaccines in which the hydrophilic MUC1 antigen was N-terminally conjugated to one or two palmitoyl lipid chains (to form amphiphilic Pam-MUC1 or Pam2 -MUC1). These amphiphilic lipid-tailed MUC1 antigens were self-assembled into liposomes containing the NKT cell agonist αGalCer as an adjuvant. The lipid-conjugated antigens reshaped the physical and morphological properties of liposomal vaccines. Promising results showed that the anti-MUC1 IgG antibody titers induced by the Pam2 -MUC1 vaccine were more than 30- and 190-fold higher than those induced by the Pam-MUC1 vaccine and the MUC1 vaccine without lipid tails, respectively. Similarly, vaccines with the TLR1/2 agonist Pam3 CSK4 as an adjuvant also induced conjugated lipid-dependent immunological responses. Moreover, vaccines with the αGalCer adjuvant induced significantly higher titers of IgG antibodies than vaccines with the Pam3 CSK4 adjuvant. Therefore, the non-covalent assembly of the amphiphilic lipo-MUC1 antigen and the NKT cell agonist αGalCer as a glycolipid adjuvant represent a synthetically simple but immunologically effective approach for the development of anti-MUC1 cancer vaccines.

A dynamic discount pricing strategy for viral marketing.

Viral marketing has been one of the main marketing modes. However, theoretical study of viral marketing is still lacking. This paper focuses on the problem of developing a cost-effective dynamic discount pricing strategy for a viral marketing campaign. First, based on a novel word-of-mouth propagation model, we model the original problem as an optimal control problem. Second, we show that the optimal control problem admits an optimal control and present the optimality system for solving the optimal control problem. Next, we solve some optimal control models to get their respective optimal dynamic discount pricing strategies. Finally, we examine the effect of some factors on the maximum marketing profit. These results contribute to gaining insight into viral marketing.

Otocephaly.

Otocephaly is a rare lethal syndrome of microstomia, aglossia, agnathia, and synotia. This male infant was born to a 19-year-old, gravida 1, para 0, woman who received routine prenatal check-up. Polyhydramnios, low-lying ears, and proboscis were noted by sonography at 29 weeks of gestation. Amniocentesis showed a normal karyotype of 46, XY. Premature rupture of membranes and preterm labor were noted at 32 weeks of gestation. A male infant was delivered preterm and died shortly after birth. The infant showed midline proboscis and absence of mandible. The simple, soft ears were extremely low-set and were near the midline of the neck. Otocephaly is regarded as the most severe form of first arch anomalies. Prenatal diagnosis should be dependent on ultrasound analysis. In the face of polyhydramnios, otocephaly is one of the possible fetal anomalies.

Targeted multidrug delivery system to overcome chemoresistance in breast cancer.

Chemotherapy has been widely used in breast cancer patients to reduce tumor size. However, most anticancer agents cannot differentiate between cancerous and normal cells, resulting in severe systemic toxicity. In addition, acquired drug resistance during the chemotherapy treatment further decreases treatment efficacy. With the proper treatment strategy, nanodrug carriers, such as liposomes/immunoliposomes, may be able to reduce undesired side effects of chemotherapy, to overcome the acquired multidrug resistance, and to further improve the treatment efficacy. In this study, a novel combinational targeted drug delivery system was developed by encapsulating antiangiogenesis drug bevacizumab into liposomes and encapsulating chemotherapy drug doxorubicin (DOX) into immunoliposomes where the human epidermal growth factor receptor 2 (HER2) antibody was used as a targeting ligand. This novel combinational system was tested in vitro using a HER2 positive and multidrug resistant breast cancer cell line (BT-474/MDR), and in vivo using a xenograft mouse tumor model. In vitro cell culture experiments show that immunoliposome delivery led to a high cell nucleus accumulation of DOX, whereas free DOX was observed mostly near the cell membrane and in cytoplasm due to the action of P-gp. Combining liposomal bevacizumab with immunoliposomal DOX achieved the best tumor growth inhibition and the lowest toxicity. Tumor size decreased steadily within a 60-day observation period indicating a potential synergistic effect between DOX and bevacizumab through the targeted delivery. Our findings clearly indicate that tumor growth was significantly delayed in the combinational liposomal drug delivery group. This novel combinational therapy has great potential for the treatment of patients with HER2/MDR double positive breast cancer.

A Biomimetic Microfluidic Tumor Microenvironment Platform Mimicking the EPR Effect for Rapid Screening of Drug Delivery Systems.

Real-time monitoring of tumor drug delivery in vivo is a daunting challenge due to the heterogeneity and complexity of the tumor microenvironment. In this study, we developed a biomimetic microfluidic tumor microenvironment (bMTM) comprising co-culture of tumor and endothelial cells in a 3D environment. The platform consists of a vascular compartment featuring a network of vessels cultured with endothelial cells forming a complete lumen under shear flow in communication with 3D solid tumors cultured in a tumor compartment. Endothelial cell permeability to both small dye molecules and large liposomal drug carriers were quantified using fluorescence microscopy. Endothelial cell intercellular junction formation was characterized by immunostaining. Endothelial cell permeability significantly increased in the presence of either tumor cell conditioned media (TCM) or tumor cells. The magnitude of this increase in permeability was significantly higher in the presence of metastatic breast tumor cells as compared to non-metastatic ones. Immunostaining revealed impaired endothelial cell-cell junctions in the presence of either metastatic TCM or metastatic tumor cells. Our findings indicate that the bMTM platform mimics the tumor microenvironment including the EPR effect. This platform has a significant potential in applications such as cell-cell/cell-drug carrier interaction studies and rapid screening of cancer drug therapeutics/carriers.

The Long-Term Effective Rate of Different Branches of Idiopathic Trigeminal Neuralgia After Single Radiofrequency Thermocoagulation: A Cohort Study.

To evaluate the efficacy of computed tomography (CT) guided single radiofrequency thermocoagualtion (RFT) in 1137 patients with idiopathic trigeminal neuralgia after a follow-up period of 11 years, specially focused on duration of pain relief in different branches of trigeminal nerve, side effect, and complications. Retrospective study of patients with idiopathic trigeminal neuralgia treated with a single CT guided RFT procedure between January 2002 and December 2013. The mean follow-up time was 46.14 ±â€Š30.91 months. Immediate postprocedure pain relief was 98.4%. V2 division obtained the best pain relief rate: 91%, 89%, 80%, 72%, 60%, and 54% at 1, 3, 5, 7, 9, and 11 years, respectively. No statistical difference pairwise comparison was in other groups. The complications included masseter muscle weakness, corneitis, diplopia, ptosis, hearing loss, limited mouth opening, and low pressure headache. Masticatory weakness mostly occurred in patients with V3 branch involvement, while Corneitis and Diplopia all in patients with V1 branch involvement. No mortalities observed during or after RFT. All different branches division of trigeminal neuralgia achieved comparable satisfactory curative effect; V2 obtained the best excellent pain relief, after RFT procedure. Facial numbness is inevitable after RFT, which patients who have pain in all 3 trigeminal divisions and patients who desire no facial numbness should be cautious. Masticatory weakness is mainly related with V3 injured, while Corneitis and Diplopia in patients with V1 injured by RFT.

Ion-Responsive Channels of Zwitterion-Carbon Nanotube Membrane for Rapid Water Permeation and Ultrahigh Mono-/Multivalent Ion Selectivity.

The rational combination of polymer matrix and nanostructured building blocks leads to the formation of composite membranes with unexpected capability of selectivity of monovalent electrolytes and water, which affords the feasibility to effeciently remove harmful ions and neutral molecules from the environment of concentrated salines. However, the multivalent ion rejection in salined water of routine nanocomposite membranes was less than 98% when ion strength is high, resulting in a poor ion selectivity far below the acceptable value. In this contribution, the ion-responsive membrane with zwitterion-carbon nanotube (ZCNT) entrances at the surface and nanochannels inside membrane has been proposed to obtain ultrahigh multivalent ion rejection. The mean effective pore diameter of ZCNT membrane was dedicated tuned from 1.24 to 0.54 nm with the rise in Na2SO4 concentration from 0 to 70 mol m(-3) as contrary to the conventional rejection drop in carbon nanotube (CNT) membrane. The ultrahigh selective permeabilities of monovalent anions against divalent anions of 93 and against glucose of 5.5 were obtained on ZCNT membrane, while such selectivities were only 20 and 1.6 for the pristine CNT membrane, respectively. The ZCNT membranes have potential applications in treatment of salined water with general NaCl concentration from 100 to 600 mol m(-3), which are widely applicable in desalination, food, and biological separation processes.

The role of surface chemistry in determining in vivo biodistribution and toxicity of CdSe/ZnS core-shell quantum dots.

To examine the effect of surface chemistry and surface charge on in vivo biodistribution and toxicity of CdSe/ZnS core-shell quantum dots (QDs), QDs with positive, negative, or PEG coating are used in this study for in vivo evaluation in a mouse model. The results suggest that QDs coated with cationic polydiallyldimethylammonium chloride (PDDA) preferentially deposit in the lung other than in the liver, while the negative and PEGylated QDs render abundant accumulation in the liver. At higher doses positive QDs with PDDA coating show severe acute toxicity due to pulmonary embolism. Independent of their surface coatings, all QDs cause injuries in specific tissues like liver, spleen, lung, and kidney, after acute and long-term exposure, and the degree of injuries is dominated by their surface properties. For the positively charged QDs, the acute phase toxicity is primarily contributed by the coating material PDDA, while coating on QDs may amplify both in vitro and in vivo toxicity of PDDA. PEGylated QDs display the slightest chronic injuries in the long-term toxicity examination in comparison to positive or negative ones.

Nanocomplexation-assisted solubilization of pDNA in organic solvents for improved microencapsulation.

With an attempt to solubilize plasmid DNA (pDNA) in organic solvents, we developed nanocomplexation strategy by using block copolymer of polyethylene glycol-b-poly(L-lysine) (PEG-b-PLL). Self-assembly of PEG-b-PLL and pDNA in aqueous solution led to spherical nanocomplexes with core-shell structure when the +/- charge ratio (N/P) was higher than 1:1. This complexation had no significant impact on the helical structure of pDNA. After lyophilization, PEG-b-PLL/pDNA nanocomplexes originally formed in aqueous solution could be well dispersed in organic solvents such as THF and dichloromethane. Facilitating by nanocomplexation, pDNA could be encapsulated into PLGA nanoparticles with high efficiency, via oil-in-water emulsion solvent evaporation method. This complexation mediated pDNA solubilization in organic solvents may facilitate the development of novel nano- and micro-platforms for gene delivery.

Nanoplexes for cell imaging and hyperthermia: in vitro studies.

Novel IR820-polyethylene glycol-diamine nanoplexes (IR820-PDNCs) have potential multifunctional imaging-hyperthermia applications in cancer. Nanoplexes were formulated by ionic interaction and characterized in vitro for their imaging and hyperthermia capabilities. The resulting nanoplexes were approximately 50 nm diameter, with a zeta potential of 2.0 +/- 0.9 mV, and able to generate heat upon exposure to 808 nm laser. Cytotoxicity studies in SKOV-3, MES-SA and Dx5 cancer cell lines demonstrate comparable cytotoxicity of IR820-PDNCs versus free IR820 after 24 hours. The nanoplexes are able to produce hyperthermic cell growth inhibition in all three cancer cell lines after excitation with laser. The level of cell growth inhibition caused by hyperthermia is significantly higher for IR820-PDNCs compared to IR820 in MES-SA and Dx5 cells. Fluorescent microscope images after 2.5-hour exposure to 5 microM IR820-PDNCs or 5 microM free IR820 show increased uptake for IR820-PDNCs compared to free IR820, especially for SKOV-3 and Dx5 cancer cells. This formulation can potentially be used in multifunctional cancer theranostics.

Applying Taguchi methods for solvent-assisted PMMA bonding technique for static and dynamic micro-TAS devices.

This work examines numerous significant process parameters in the solvent-assistant Polymethyl methacrylate (PMMA) bonding scheme and presents two Micro-total-analysis System (micro-TAS) devices generated by adopting the optimal bonding parameters. The process parameters considered were heating temperature, applied loading, duration and solution. The effects of selected process parameters on bonding dimensions loss and strength, and subsequent optimal setting of the parameters were accomplished using Taguchi's scheme. Additionally, two micro-TAS devices were realized using a static paraffin microvalve and a dynamic diffuser micropump. The PMMA chips were carved using a CO2 laser that patterned device microchannels and microchambers. The operation principles, fabrication processes and experimental performance of the devices are discussed. This bonding technique has numerous benefits, including high bonding strength (240 kgf/cm2) and low dimension loss (2-6%). For comparison, this work also demonstrates that the normal stress of this technology is 2-15 times greater than that of other bonding technologies, including hot embossing, anodic bonding, direct bonding and thermal fusion bonding.