A handheld continuous-flow real-time fluorescence qPCR system with a PVC microreactor.

The polymerase chain reaction (PCR) has unique advantages of sensitivity, specificity and rapidity in pathogen detection, which makes it at the forefront of academia and application in molecular biology diagnosis. In this study, we proposed a hand-held real-time fluorescence qPCR system, which can be used for the quantitative analysis of nucleic acid molecules. For the first time, we use a PVC microreactor which improved the transmittance of the microreactor and made it easy to collect the fluorescence signal. In order to make it portable, the system adopted a passive syringe for sample injection and integrated temperature control and detection with a lithium battery for power supply. What's more, the fluorescence signal was captured by using a smartphone through an external automatic robotic arm. This real-time qPCR system can detect genomic DNA of the H7N9 avian influenza over four orders of magnitude of concentration from 107 to 104 copies per µL. In addition, it was verified that the fluorescence images obtained by this system were clearer than those obtained by a traditional system (using a PTFE spatial PCR microreactor) with two typical dyes and a probe tested-EvaGreen, SYBR Green and FAM.

Flow-through polymerase chain reaction inside a seamless 3D helical microreactor fabricated utilizing a silicone tube and a paraffin mold.

We introduce a new strategy for fabricating a seamless three-dimensional (3D) helical microreactor utilizing a silicone tube and a paraffin mold. With this method, various shapes and sizes of 3D helical microreactors were fabricated, and a complicated and laborious photolithographic process, or 3D printing, was eliminated. With dramatically enhanced portability at a significantly reduced fabrication cost, such a device can be considered to be the simplest microreactor, developed to date, for performing the flow-through polymerase chain reaction (PCR).

Bronchobiliary fistula after traumatic liver rupture: a case report.

INTRODUCTION: Bronchobiliary fistulas are rare and difficult to treat. Peacock first reported this entity in 1850 while treating a patient with hepatic encopresis. CASE PRESENTATION: A 67-year-old Chinese male patient presented to the outpatient clinic with a complaint of coughing up phlegm with chest tightness for 4 days with symptoms of intermittent bilirubin sputum with a sputum volume of about 500 ml per day but no symptoms of abdominal pain or jaundice and no yellow urine or steatorrhea. The examination revealed cyanosis of the lips and mouth, barrel chest, low breath sounds on the right side, and a large number of wet rales heard in both lungs. The imaging investigations were suggestive of bronchobiliary fistula. Therefore, the patient was operated on and discharged with no perioperative complications. CONCLUSION: Bronchobiliary fistula should be considered diagnostically in patients with known liver disease who also experience trauma or medical treatment and cough up bile-colored sputum, regardless of the presence of concurrent infections, and in conjunction with radiological expertise to identify it. Here, we report a case of bronchobiliary fistula and a brief review of the literature on it.

Flow-through PCR on a 3D qiandu-shaped polydimethylsiloxane (PDMS) microdevice employing a single heater: toward microscale multiplex PCR.

Consistent temperature control in an on-chip flow-through polymerase chain reaction (PCR) employing two or more heaters is one of the main obstacles for device miniaturization and integration when realizing micro total analysis systems (µTAS), and also leads to operational complexity. In this study, we propose a qiandu (right triangular prism)-shaped polydimethylsiloxane (PDMS) microdevice with serpentine microchannels fabricated on its slanted plane, and apply the device for an on-chip flow-through PCR employing a single heater. The inclined nature of the qiandu-shaped microdevice enables the formation of a surface temperature gradient along the slanted plane of the microdevice in a height-dependent manner by the use of a single heater, and enables liquid to traverse over wide ranges of temperatures, including the three temperature zones--denaturation, annealing, and extension temperatures--required in a typical PCR. The feasibility of the qiandu-shaped PDMS microdevice as a versatile platform for performing a flow-through PCR was examined by employing multiple templates and varying the inclination angle of the device. In addition, the potential of performing a multiplex PCR using a single qiandu-shaped PDMS microdevice was explored. A 409 bp long gene fragment effective as a marker for diagnosing lung cancer and a 230 bp long gene fragment from a plasmid vector were simultaneously amplified in less than 25 min on a single microdevice, paving the way for a microscale, multiplex PCR on a single device employing a single heater.

Prognostic effects of surgery or radiotherapy on adenoid cystic carcinoma of the head and neck: a retrospective cohort study.

Background: There is little evidence exploring prognostic effects of surgery and radiotherapy on adenoid cystic carcinoma (ACC) of the head and neck. This study sought to evaluate the prognostic effects of surgery or radiotherapy on ACC of the salivary gland, oropharynx, and nose, nasal cavity, and middle ear. Methods: In this cohort study, the data of 2,392 participants with ACC of the head and neck were extracted from the Surveillance, Epidemiology and End Results (SEER) database. Participants were divided into the salivary gland group (n=1,351), the mouth and oropharynx group (n=563), and the nose, nasal cavity, and middle ear group (n=478). Baseline characteristics were assessed via questionnaires or laboratory analysis and outcome variables were all-cause death and cancer-specific death of patients. Baseline data were collected in 2004, and patients were followed-up to 2016. The survival time of patients were recorded. Univariate and multivariate Cox regression analyses explored the effects of surgery and radiotherapy on overall prognosis of ACC patients. Fine-Gray test assessed the effects of surgery and radiotherapy on cancer-specific mortality of ACC patients. Results: In total, 766 died and 1,626 survived with a median survival time of 9.92 years. After adjusting for confounders, patients with ACC of the salivary gland who underwent surgery had a decreased risk of all-cause mortality [hazard ratio (HR) =0.51; 95% confidence interval (CI): 0.36-0.71] and cancer-specific mortality (HR =0.57; 95% CI: 0.34-0.97). Surgery was found to be a protective factor for the risk of all-cause mortality (HR =0.47; 95% CI: 0.28-0.78) and cancer-specific mortality (HR =0.70; 95% CI: 0.33-1.50) of patients with ACC of the mouth and oropharynx after adjusting for confounders. After adjusting for confounders, patients with ACC of the nose, nasal cavity, and middle ear who underwent surgery had a decreased risk of all-cause mortality (HR =0.46; 95% CI: 0.30-0.70) and cancer-specific mortality (HR =0.35; 95% CI: 0.20-0.61). Conclusions: Surgery was associated with a decreased risk of mortality in patients with ACC of the salivary gland, oropharynx, and nose, nasal cavity, and middle ear, which suggested the value of surgery for improving their prognosis.

Three-dimensional on-chip continuous-flow polymerase chain reaction employing a single heater.

Multi-step temperature control in a polymerase chain reaction (PCR) is a limiting factor in device miniaturization and portability. In this study, we propose the fabrication of a three-dimensional (3D) microdevice employing a single heater to minimize temperature control required for an on-chip continuous-flow PCR as well as the overall footprint by stacking the device in multi-layers. Two poly(dimethylsiloxane) (PDMS) layers with differing thicknesses are vertically stacked with their microchannel-engraved sides facing down. Through-holes are made in the thicker PDMS layer, which is sandwiched between a glass substrate at the bottom and the thinner PDMS layer at the top. In this way, a fluidic conduit is realized in a 3D configuration. The assembled 3D microdevice is then placed onto a heater glass-side down. The interface of the two PDMS layers displays a relatively lower temperature than that of the PDMS and glass layers due to the low thermal conductivity of the PDMS and its physical distance from the heater. The denaturation temperature can be controlled by adjusting the temperature of the heater, while the annealing/extension temperature can be controlled automatically by molding the thicker bottom PDMS layer into the appropriate thickness calculated using a numerical derivation proposed in this study. In this way, a cumbersome temperature measurement step is eliminated. DNA amplification was successfully carried out using the proposed 3D fluidic microdevice, and the intensity of the resulting amplicon was comparable to that obtained using a thermal cycler. This novel concept of adopting a single heating source greatly simplifies the temperature control issue present in an on-chip continuous-flow PCR. It also allows the use of a commercialized hot plate as a potential heat source, paving the way for device miniaturization and portability in a highly cost-effective manner. In this study, a simple and facile technique to make arrays of through-holes for the fluidic interconnection inside a 3D channel configuration is also addressed.

[Mechanical complications with self-made, antibiotic-loaded cement articulating spacers in the treatment of the infected hip replacement].

OBJECTIVE: To study the complications and efficacy of self-made, antibiotic-loaded cement articulating spacers in the treatment of the infected hip replacement. METHODS: Between January 2006 and July 2016, 265 patients (266 hips) received a self-made, antibiotic-loaded cement articulating spacer as part of a two-stage protocol. Among those patients, there were 143 males(144 hips) and 122 females(122 hips). The cement articulating spacers with vancomycin and two Steinman pins were made by a self-made mold system. Meanwhile, another antibiotic was added to the spacers according to the drug sensitivity test. Record if the infected prosthesis was removed, related complication with spacer(breakage and dislocation), Harris score, and control rate of infection. RESULTS: The mean age of two-stage revision operation was(57.4±14.2) years. Thirty-nine patients(14.7%) used extended trochanteric osteotomy(ETO) to remove the infected prosthesis. And 38 patients occurred mentioned complications(14.3%). Spacer breakage occurred in 28 cases(10.5%) and dislocation occurred in 10 cases(3.8%). The mean follow-up time was(83.4±14.6) months. The Harris hip score was from 47.56±14.23 preoperatively to 86.43±12.84 at final follow-up(P<0.05). The infection of 256 cases(96.6%) got control after revision operation. However, during postoperative follow-up, 4 cases occurred re-infection, and they were reoperated, and the infections obtained effective control after the operation. Thus total infection control rate was 95.1%(252/265). CONCLUSIONS: Antibiotic-loaded cement articulating spacer made by a self-made mold system is effective in controling infection caused by hip replacement. Related complication is less with spacer by a mould enclosing two Steinman pins. Using metallic internal fixation or allograft bone combined with spacer does not affect infection control.

Instantaneous room temperature bonding of a wide range of non-silicon substrates with poly(dimethylsiloxane) (PDMS) elastomer mediated by a mercaptosilane.

This paper introduces an instantaneous and robust strategy for bonding a variety of non-silicon substrates such as thermoplastics, metals, an alloy, and ceramics to poly(dimethylsiloxane) (PDMS) irreversibly, mediated by one-step chemical modification using a mercaptosilane at room temperature followed by corona treatment to realize heterogeneous assembly also at room temperature. The mercapto functional group is one of the strongest nucleophiles, and it can instantaneously react with electrophiles of substrates, resulting in an alkoxysilane-terminated substrate at room temperature. In this way, prior oxidation of the substrate is dispensed with, and the alkoxysilane-terminated substrate can be readily oxidized and irreversibly bonded with oxidized PDMS at room temperature. A commercially available Tesla coil was used for surface oxidation, replacing a bulky and expensive plasma generator. Surface characterization was conducted by water contact angle measurement and X-ray photoelectron spectroscopy (XPS) analysis. A total of fifteen non-silicon substrates including polycarbonate (PC), two types of poly(vinylchloride) (PVC), poly(methylmethacrylate) (PMMA), polystyrene (PS), polyimide (PI), two types of poly(ethylene terephthalate) (PET), polypropylene (PP), iron (Fe), aluminum (Al), copper (Cu), brass, alumina (Al2O3), and zirconia (ZrO2) were bonded successfully with PDMS using this method, and the bond strengths of PDMS-PMMA, PDMS-PC, PDMS-PVC, PDMS-PET, PDMS-Al, and PDMS-Cu assemblies were measured to be approximately 335.9, 511.4, 467.3, 476.4, 282.2, and 236.7 kPa, respectively. The overall processes including surface modification followed by surface oxidation using corona treatment for bonding were realized within 12 to 17 min for most of the substrates tested except for ceramics which required 1 h for the bonding. In addition, large area (10 × 10 cm(2)) bonding was also successfully realized, ensuring the high reliability and stability of the introduced method.

[Clinical features of inflammatory myofibroblastic tumor in the head and neck].

OBJECTIVE: To summarize the clinical characteristics of inflammatory myofibroblastic tumor (IMT) in head and neck and to discuss its treatment. METHODS: Twenty-seven cases of IMT in head and neck diagnosed at the Chinese PLA General Hospital from 2004 to 2012 were analyzed retrospectively. Among the 27 patients, 12 males and 15 females, age ranged from 8 to 77 years, with a median 43 years old. Treatment included: 1 with radiotherapy, 22 with surgery, 3 with surgery and postoperative radiotherapy, one with concurrent chemoradiotherapy followed by surgery. Of the 27 cases, 5 located in the neck, 6 in the nasal and paranasal sinus, 4 in the temporal bone, 3 in the throat, 2 in the parotid gland, 2 in the lower pharynx, 1 in the mandible, 1 in the maxilla, 1 in the masseter muscle, 1 in the amygdala and 1 in the pharynx nasalis. RESULTS: Following-up time was 4-85 months, with a median of 26 months. Six cases lost follow-up, 1 case with malignant transformation and died, 16 cases survived with no recurrence, 4 cases relapsed, of whom 2 were alive with tumors and 2 died. CONCLUSIONS: IMT in the head and neck has a tendency to be malignancy, with certain recurrence rate and mortality. Radical excision is still the first choice of treatment for IMT in head and neck.

[Research of postoperative quality of life of laryngeal carcinoma patients].

OBJECTIVE: To explore and compare the effect of clinical characters on quality of life (QOL) of laryngeal carcinoma postoperative patients. METHOD: We performed a retrospective cohort follow up study of 303 patients who underwent operation for laryngeal carcinoma. The Chinese version of the general and the head and neck specific quality of life questionnaires of the European Organization for the Research and Treatment of Cancer (EORTC QLQ-C30 and EORTC QLQ-H&H35) were used to measure the quality of life of those patients in the cohort, to study the relationship between QOL and clinical factors. RESULT: In the 303 cases, there were 78 cases retaining the permanent tracheostomies (59 total laryngectomy and 19 partial laryngectomy); 4 patient retain the nasal feed pipe. Female patients were worse than male patients in the domain of physical function and global QOL (P<0.05). Older patients were worse than younger patients in the domain of physical function and speech (P<0.05). Patients with glottic tumors experience better global QOL than their peers with under glottic or supraglottic tumors. The patients with supraglottic tumor have more problem with swallowings (P<0.05). Earlier stage patients have better suitation in many domains than later stage patients, but III stage have more difficulty with swallowing than IV stage. Patients who accepted minimally invasive operation and laryngofissure have better QOL than those accepted partial laryngectomy or total laryngectomy; total laryngectomee have more problems with physical, emotion function, global QOL, sense, speech, cough and morbid feeling than patients with partial laryngectomy, but less problem with fatigue, pain, dyspnea, swollowing and dry mouth; Patients with partial horizontal laryngectomy have more difficulty with swallowing. In the multivariable analysis, the operation mode was an independent factor to speech problem. Patients who accepted neck dissction have more negative outcomes than patients without neck dissection. In the univariable analysis, the permanent tracheostomy was a notable factor which affected many domains in the quality of life; multivariable analysis showed that permanent tracheostomy was an important factor which worked on global quality of life. CONCLUSION: The quality of life was affected by many clinical factors. The operation mode was an independent factor which affect speech problem. Decannulation after partial laryngectomy was very important to keep the quality of life.

[Diagnosis and treatment of the primary parapharyngeal space tumors].

OBJECTIVE: To study the clinical characteristics, diagnosis and surgical managements of the parapharyngeal space tumors. METHODS: A retrospective study of 40 patients with primary parapharyngeal space tumors treated from January 2006 to December 2008 in Chinese PLA General Hospital was performed. Among the 40 patients, there were male 22 patients, female 18 (45%), age ranged from 1 - 77, median 42 years old. CT scan combined with MRI was helpful to diagnose the parapharyngeal space tumor and make surgical plan. The surgical approaches include: trans-oral in 1 patient, trans-cervical approach in 22, transcervical-parotid approach in 8, vertical ramus osteotomy approach in 1, transcervical-partial bone resection in the angle of mandible in 4, transparotid approach in 2, and transcervical in combination with post auricle craniotomy approach in 2. RESULTS: All 40 patients had undergone surgical treatment. Postoperative histopathology showed benign in 28 patients and malignant in 12 patients. The tumors originating from salivary glands were in 15 patients, neurogenic tumors in 12 patients and tumors originating from other tissues were in 13 patients.Among 28 patients with benign tumors, 23 had been cured with one operation, without recurrence during following-up of 13 - 47 months, with a median of 39 months. Among 12 patients with malignant tumors, 6 patients alive (with following-up of 24 - 50 months and a median of 36 months), 3 patients died in half year after operation and 3 patients lost. The post-operative complication included Cerebrospinal fluid leak in one patient, operative field infection in 2 patients, and vagus nerve injury in 3 patients. CONCLUSIONS: Surgery is the first choice for parapharyngeal space tumors. Transcervical approach alone can apply to most tumors and a broader approach is indicated for malignant or large benign tumors. The prognosis is good for the benign lesions, but poor for the malignant tumors.

Diagnosis and treatment of giant cell granuloma of the temporal bone: Report of eight cases.

UNLABELLED: Abstract Conclusions: It is impossible to make a diagnosis of temporal giant cell granuloma (GCG) before operation because of nonspecific clinical and imaging feature. Surgery is the first-line choice of treatment. OBJECTIVE: To evaluate the diagnosis and treatment of temporal GCG. METHODS: Eight patients with GCG receiving treatment in the Chinese PLA General Hospital between 2001 and 2010 were recruited for the study. These patients' clinical features, imaging and histopathological findings, types of surgery, and results of follow-up evaluations were noted. RESULTS: The group was made up of four males and four females, with a median age of 37 years (range 21-50 years). Four patients had a granuloma on the left side and four on the right of the head. The median duration of the disease was 21 months (range 5-60 months). All patients, except one referred to us for recurring disease, were managed in our hospital. The main symptoms were: hearing loss (n = 5), tinnitus (n = 4), otalgia (n = 3), dizziness (n = 2), and local masses (n = 2). Radiological examination of the masses revealed erosion of the temporal bone and base of the skull. There was no definitive diagnosis in any of the patients before surgery. All patients had surgical treatment, six of them by middle cranial fossa approach and two by combined cranio-auricular approach. Surgical complications included partial facial paralysis (three cases) and cerebral edema (one case) but they resolved soon after surgery. One patient was lost to follow-up, but the other seven were followed up over a mean period of 24 months; none has reported a recurrence.

[Observe the curative effect of n-Butyl cyanoacrylate injection in treating laryngopharynx hemangioma].

OBJECTIVE: To evaluate the clinical effectiveness of injection n-Butyl cyanoacrylate (NBCA) in treating laryngopharynx hemangioma. METHOD: Thirty cases of laryngopharynx hemangioma who received NBCA injection were obtained from our department 1998-2007. Twenty-five cases had tracheotomy under General anesthesia, and NBCA was injected into hemangioma by direct laryngoscopy. NBCA was mixed with iodide, and the concentration was 25.00% to 33.33%, NBCA dosage was 0.5 ml to 2.0 ml. There were 5 cases whose hemangioma confined to oropharynx didn't have tracheotomy, and they were injected straightly with the speculum oris. We observed the shedding of hemangioma in different time. RESULT: Hemangioma with diameter less than 1 cm, shedded in about 1 month. However, hemangioma with diameter more than 4 cm, shedded in a period more than 3.5 month. No recurrence was observed in the follow up of 3 months to seven years. CONCLUSION: Avoiding dissection of neck and repetitious operation, NBCA injection could be a safe, simple and effective therapy for laryngopharynx hemangioma.