Optimized nickase- and nuclease-based prime editing in human and mouse cells.

Precise genomic modification using prime editing (PE) holds enormous potential for research and clinical applications. In this study, we generated all-in-one prime editing (PEA1) constructs that carry all the components required for PE, along with a selection marker. We tested these constructs (with selection) in HEK293T, K562, HeLa and mouse embryonic stem (ES) cells. We discovered that PE efficiency in HEK293T cells was much higher than previously observed, reaching up to 95% (mean 67%). The efficiency in K562 and HeLa cells, however, remained low. To improve PE efficiency in K562 and HeLa, we generated a nuclease prime editor and tested this system in these cell lines as well as mouse ES cells. PE-nuclease greatly increased prime editing initiation, however, installation of the intended edits was often accompanied by extra insertions derived from the repair template. Finally, we show that zygotic injection of the nuclease prime editor can generate correct modifications in mouse fetuses with up to 100% efficiency.

[Two cases of herpes simplex keratitis after trans-epithelial photorefractive keratectomy].

We herein report 2 cases of herpes simplex keratitis after trans-epithelial photorefractive keratectomy. Patients' medical histories, symptoms, signs, clinical examination results, diagnosis and treatment were showed in detail. Following precision diagnosis and medical intervention, including topical and systemic antiviral treatmented for 1 to 2 weeks. The two patients were cured with full reepithelialization without corneal scar.

[Efficacy of mitomycin C 0.02% for prevention of haze after transepithelial photorefractive keratectomy for mild and moderate myopia].

Objective: To investigate the efficacy of mitomycin C (MMC) 0.02% for prevention of haze after transepithelial photorefractive keratectomy (Trans-PRK) for mild and moderate myopia. Methods: Retrospective cohort study. We reviewed medical records of 295 patients (588 eyes) who underwent Trans-PRK with or without use of MMC. There were 45 patients (90 eyes) in the mild myopia group (aged between 18 and 41 years; 37 males and 8 females; myopia diopter <3.00 D) and 250 patients (498 eyes) in the moderate myopia group (aged between 18 and 46 years; 168 males and 82 females; myopia diopter: 3.00 to 6.00 D). The two groups were divided into subgroups with MMC 0.02% and without MMC, respectively. The time of intraoperative application of MMC, if there was, was 15 s and 30 s in the mild myopia group and the moderate myopia group, respectively. The mean follow-up time was 6 months. Postoperative best corrected visual acuity (BCVA), spherical equivalent (SE) and haze were analyzed and compared using an independent Student t-test or Mann-Whitney U test between subgroups. Haze variables were compared using chi-square statistics. Results: Haze was quantified with Fantes from grade 0.5 to 4. In the mild myopia group, all haze grades were 0.5 within 3 months. The incidence of haze was 6.25% (2/32) in eyes treated with MMC and 8.62% (5/58) in eyes treated without MMC; there was no statistical significance (χ²=0.00, P>0.999). In the moderate myopia group, the incidence of haze was 9.19% (24/261) in eyes treated with MMC within 3 months; the grade was 0.5 in 91.67% (22/24) of eyes with haze and 1 in 8.33% (2/24). The incidence of haze was 29.53% (70/237) in eyes treated without MMC; the grade was 0.5 in 60.00% (42/70) of eyes with haze, 1 in 18.57% (13/70), and 2 in 5.71% (4/70) within 3 months, and 0.5 in 15.71% (11/70) after 3 months (χ²=12.36, P=0.002). In the mild myopia group, BCVA was 5.0(5.0, 5.1) versus 5.0(5.0, 5.1) in the subgroups with MMC and without MMC (Z=-0.34, P=0.733). In the moderate myopia group, BCVA was 5.0(5.0, 5.1) versus 5.0(5.0, 5.1) in the subgroups with and without MMC (Z=-2.05, P=0.040). In the mild myopia group, SE was (0.33±1.07) D versus (0.32±0.57) D in the subgroups with and without MMC (t=0.25, P=0.805). In the moderate myopia group, SE was (0.66±0.85) D versus (0.53±0.67) D in the subgroups with and without MMC (t=2.97, P=0.003). Conclusions: MMC 0.02% was effective in preventing haze after Trans-PRK in the treatment of moderate myopia. However, it was not effective in mild myopia.

Nivolumab with carboplatin, paclitaxel, and bevacizumab for first-line treatment of advanced nonsquamous non-small-cell lung cancer.

BACKGROUND: This international, randomized, double-blind phase III study (ONO-4538-52/TASUKI-52) evaluated nivolumab with bevacizumab and cytotoxic chemotherapy as first-line treatment for nonsquamous non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: Between June 2017 and July 2019, this study enrolled treatment-naïve patients with stage IIIB/IV or recurrent nonsquamous NSCLC without sensitizing EGFR, ALK, or ROS1 alterations. They were randomly assigned in a 1 : 1 ratio to receive nivolumab or placebo in combination with carboplatin, paclitaxel, and bevacizumab every 3 weeks for up to six cycles, followed by nivolumab/placebo with bevacizumab until progressive disease or unacceptable toxicity. The primary endpoint was progression-free survival (PFS) assessed by an independent radiology review committee (IRRC). RESULTS: Overall, 550 patients from Japan, Korea, and Taiwan were randomized; of these patients, 273 and 275 received the nivolumab and placebo combinations, respectively. In the present preplanned interim analysis with a median follow up of 13.7 months, the IRRC-assessed median PFS was significantly longer in the nivolumab arm than in the placebo arm (12.1 versus 8.1 months; hazard ratio 0.56; 96.4% confidence interval 0.43-0.71; P < 0.0001). The PFS benefit was observed across all patients with any programmed death-ligand 1 (PD-L1) expression levels including PD-L1-negative patients. The IRRC-assessed objective response rates were 61.5% and 50.5% in the nivolumab and placebo arms, respectively. The incidence of treatment-related adverse events of grade 3 or 4 was comparable between the two arms; treatment-related adverse events leading to death were observed in five and four patients in the nivolumab and placebo arms, respectively. CONCLUSION: The TASUKI-52 regimen should be considered a viable new treatment strategy for treatment-naïve patients with advanced nonsquamous NSCLC.

[RNA interference targeting DNA-PKcs inhibits glioma cells malignancies and enhances temozolomide sensitivity].

Objective: To investigate the effect of DNA dependent protein kinase catalytic subunit (DNA-PKcs) on glioma proliferation, invasion and temozolomide sensitivity, and also to explore the potential mechanisms. Methods: Human glioma cell lines H4 and U87 were chosen to carry out RNA interference transfection, and then divided into negative control group (blank group) and siRNA group (test group). The knockdown efficacy of DNA-PKcs siRNA was tested by quantitative PCR and Western blot. The MTS assay and Transwell assay were used to investigate the effect of DNA-PKcs knockdown on glioma cell growth and invasion, respectively. We also used MTS assay to investigate the IC(50) value of temozolomide in negative control group and siRNA groups. Result: Compared with blank group, DNA-PKcs specific siRNA significantly downregulated both mRNA and protein level of DNA-PKcs. MTS assay results demonstrated that 72-hours proliferation of test group were only 52.48%, 54.70% (H4) and 52.98%, 50.45% (U87) of the blank group's counterpart. Transwell assay results showed that the invasiveness abilities of blank and test groups were 1.00±0.03, 0.41±0.05, 0.39±0.04 (H4) and 1.00±0.02, 0.28±0.04, 0.27±0.04 (U87). Moreover, knockdown of DNA-PKcs significantly decreased the temozolomide IC(50) value (H4: 249±27, 97±39, 88±35; U87: 485±41, 86±49, 73±38). Further we applied the Western blot to reveal the mechanism of inhibitory effect of DNA-PKcs knockdown on glioma malignancies and temozolomide sensitivity. We found that downregulation of DNA-PKcs reduced the activity of AKT signal and the expression of its downstream effectors, such as c-Myc, MMP9, and Survivin. Conclusion: RNA interference targeting DNA-PKcs could inhibit glioma malignancies and enhance temozolomide sensitivity. The inhibitory effect of DNA-PKcs knockdown on those biological activities were mainly through inhibition of AKT signal and its downstream effectors.

[Distribution and compensation mechanism of aberrations between anterior and posterior surface of the cornea in myopia and myopic astigmatism eyes].

Objective: To study the compensation mechanism of aberrations between anterior and posterior corneal surface, and to investigate the correlations between corneal aberrations and K values, intraocular pressure and central corneal thickness. Methods: One hundred and sixty-one subjects (300 eyes) with myopia and myopic astigmatism were recruited randomly. Corneal aberrations (anterior, posterior and total) in three different optical zones (2 mm, 4 mm, and 6 mm) centered on the corneal vertex were assessed with a corneal topographer (Sirius). We also calculated compensation factors (CFs) as a measure of the relative efficiency of the aberration compensation mechanism. Astigmatism was divided into corneal astigmatism and non-corneal astigmatism. Mann-Whitney U test was utilized for the comparison of different aberrations and CFs between the two astigmatism groups. Spearman correlation was applied to analyze the correlations between corneal aberrations and K values, intraocular pressure and central corneal thickness. Results: As the order of the aberrations elevated from the second to the seventh, the RMS values decreased significantly. The larger the optical zone, the greater the values of aberrations and the lower the percentage of the compensation mechanism among all the Zernike terms. At the same time, as the order of the aberrations increased, the predominance of the compensation mechanism was increasingly obvious. Slight compensation of spherical aberration (Z40) was observed in the peripheral. In the center of the analyzing zones (2 mm), compensation mechanism represented in coma (Z3±1), and it disappeared in the peripheral. We detected slight compensation of the corneal astigmatism (Z2±2) in the surrounding zone, although the compensation factors were closed to zero in diverse optical zones. Superposition was found in trefoil (Z3±3) straightly. Nevertheless, a tendency towards compensation was discovered with the enlargement of the optical zones. And the secondary spherical aberration (Z60) behaved compensation continuously. However, similar compensation was discovered between the right eye and left eye in different analyzing optical zones. There were significant correlations between the flattest K values and Z2±2, Z3±3and Z40 in the anterior, posterior and total cornea. It was similar with the steepest K values. When the optical zone expanded to 6 mm, Z3±1 and Z40 significantly decreased with intraocular pressure (rcoma=-0.188, P<0.05. rspherical=-0.147, P<0.05). No correlation was found between various aberrations and central corneal thickness (P>0.05). Conclusions: Compensation dominated in the corneal center, while the percentage decreased gradually as the optical zone extended. Slight compensation in astigmatism and spherical aberration between the anterior and posterior cornea may be benificial to the scotopic visual quality. Corneal aberrations were significantly related to K values and intraocular pressure. (Chin J Ophthalmol, 2016, 52: 840-849).

[An analysis of changes in posterior corneal elevation and relevant factors after small incision lenticule extraction].

OBJECTIVE: To investigate changes in posterior corneal elevation after small incision lenticule extraction (SMILE) and related factors. METHODS: Retrospective case series study. Eighty-three eyes of 44 myopic patients undergoing SMILE were examined with the Pentacam preoperatively, and at 1 day, 1 month, 3 months, and 6 months postoperatively. Posterior corneal elevation at the corneal apex and 0°, 45°, 90°, 135°, 180°, 225°, 270° and 325° points of the 2 mm and 6 mm diameter (total, 17 points) was analyzed. RESULTS: The changes in posterior corneal elevation at 1 day, 1 month, 3 months, and 6 months were(-1.72±2.59), (-0.98±2.37), (-0.45±1.81) and (-0.25±2.20) µm, respectively, at the corneal apex. The results were statistically significant (t=6.07, 3.75, 6.07; P<0.05), except 6 months. The changes in posterior corneal elevation were (-1.42±2.06),(-0.69±1.86), (-0.30±1.50) and(-0.22±1.58) µm, respectively, in the 2 mm circle. The results of 1 day and 1 month were statistically significant (t=6.28, 6.28, P<0.05). The changes in posterior corneal elevation were (1.48±1.47),(0.98±1.32),(0.90±1.31) and (0.90±1.16) µm, respectively, in the 6 mm circle .The results were totally statistically significant (t=6.28, 6.28, P<0.05). There were statistically significant differences between 1 month and 1 day postoperatively in the corneal apex, 2 mm and 6 mm circles. The changes were (0.75±2.55),(0.73±1.97) and(-0.50±1.60) µm. There were statistically significant differences between 3 months and 1 month postoperatively in the corneal apex and 2 mm circle. The changes were (0.53±2.22) and (0.39±1.80) µm. No significant change was found in the 6 mm circle. Between postoperative 6 months and 3 months, there were no statistically significant differences. The spherical equivalent, intraocular pressure, ablation depth, residual bed thickness, corneal hysteresis, and corneal resistance factor had no obvious correlation with the changes in posterior corneal elevation. CONCLUSIONS: After SMILE, the surrounding cornea was slightly forward, while the central posterior cornea was slightly backwards, and returned gradually. The spherical equivalent, intraocular pressure, ablation depth, residual bed thickness, corneal hysteresis, and corneal resistance factor had no obvious correlation with the changes in posterior corneal elevation. (Chin J Ophthalmol, 2016, 52: 494-498).

[Comparison of the anterior corneal asphericity after small incision lenticule extraction and femtosecond laser in situ keratomileusis].

OBJECTIVE: To evaluate and compare the anterior corneal asphericity after small incision lenticule extraction(SMILE)and femtosecond laser in situ keratomileusis(FS-LASIK). METHODS: In this case-control study, 45 subjects who underwent SMILE operation comprised the study group, and 33 subjects with FS-LASIK operation comprised the control group. The asphericity coefficient Q-value of the right eyes in both groups was measured at diameters of 6, 7, 8 and 9 mm, respectively, before surgery and at 1 day, 1 week, 1 month and 6 months following surgery. The correlation between the variation of Q-value and the central cutting depth was analyzed. RESULTS: The Q-value of anterior corneal surface was 0.85 ± 0.31, 0.80±0.28, 0.69±0.25 and 0.51±0.23 after SMILE, and 1.13±0.56, 1.01±0.47, 0.80±0.39 and 0.51±0.31 after FS-LASIK at 1 week. In both groups, the Q-value was significantly different before and after surgery(P< 0.05); there were interaction effects between the operation method and time; the difference between the two groups at 6-mm and 7-mm diameters was statistically significant(P<0.05). The variation of the Q-value before and after operation(ΔQ)showed significant difference(P6mm=0.004, P7mm=0.014)between the two groups at 6-mm and 7-mm diameters. The cap diameter of the SMILE group was smaller than that of the FS-LASIK group, but the cutting depth was larger. There was no correlation between ΔQ and the cap/disc diameter. It showed a linear relationship(P<0.05)between ΔQ and the central cutting depth at all examined diameters in the two groups, and the relation degree in the FS-LASIK group was superior to the SMILE group. CONCLUSIONS: Both SMILE and FS-LASIK operations can change the negative Q-value of the anterior corneal surface to the positive. The impact of SMILE on the asphericity is smaller than that of FS-LASIK. (Chin J Ophthalmol, 2016, 52: 681-685).

Direct observation therapy with appropriate treatment regimens was associated with a decline in second-line drug-resistant tuberculosis in Taiwan.

The resistance of Mycobacterium tuberculosis (MTB) to second-line drugs (SLDs) is growing worldwide; however, associations between the appropriateness of treatment for tuberculosis (TB) and whether the directly observed treatment, short course (DOTS)/DOTS-plus programs had an impact on the prevalence of SLD-resistant MTB are still uncertain. We performed a retrospective analysis of resistance profiles among MTB isolates obtained from 6,035 consecutive patients from 2004 to 2011 at two TB referral hospitals in Taiwan. There was a significant decrease (all p-values <0.01) in the prevalence of MTB isolates that were resistant to fluoroquinolones, injectable SLDs, and orally administered SLDs, and multidrug-resistant (MDR) and extensively drug-resistant (XDR) MTB isolates over time. There was a significant increase in the coverage rate of DOTS/DOTS-plus programs and that of administering appropriate first-line and second-line regimens (all p < 0.01). Compared with isoniazid-susceptible isolates, high-level (1.0 mg/L) isoniazid-resistant and MDR isolates showed extensive cross resistance to ofloxacin (5.9%, p < 0.01 and 33.6%, p < 0.01), levofloxacin (9.6%, p < 0.01 and 38.1%, p < 0.01), moxifloxacin (11.1%, p < 0.01 and 26.5%, p < 0.01), kanamycin (6.8 %, p < 0.01 and 16.7 %, p < 0.01), ethionamide (6.4%, p < 0.01 and 16.2%, p < 0.01), and para-aminosalicylic acid (13.1%, p < 0.01 and 20.4%, p < 0.01), but not to capreomycin (2.0%, p = 0.06 and 1.6%, p = 0.08). The decline in prevalence of resistance to SLDs was negatively correlated with the rise in rates of administering appropriate regimens as well as the DOTS/DOTS-plus programs, but not with the increase in usage of second-line regimens. The implementation of DOTS/DOTS-plus programs with appropriate regimens was associated with a decrease in the prevalence of SLD-resistant and XDR TB.

Clinical characteristics and outcomes of patients with pleural infections due to Stenotrophomonas maltophilia at a medical center in Taiwan, 2004-2012.

Stenotrophomonas maltophilia can cause various clinical diseases; however, pleural infections due to S. maltophilia are rare. We evaluated the clinical characteristics and outcomes of patients with pleural infections (complicated parapneumonic effusion or empyema) due to S. maltophilia who were treated at a medical center in Taiwan from 2004 to 2012. During the study period, 40 patients were treated for pleural infections due to S. maltophilia. The incidence of S. maltophilia pleural infections ranged from 2.66 per 1,000,000 patient-days in 2009 to 12.44 per 1,000,000 patient-days in 2011. Most of the patients with S. maltophilia pleural infections were immunocompromised male adults and all of the infections were acquired in healthcare settings. The majority of patients had polymicrobial pleural infections (n = 31, 77.5 %) and the most common pathogen was Pseudomonas aeruginosa (n = 12). The causes of pleural infections due to S. maltophilia were pneumonia due to S. maltophilia in two patients (5 %), post-surgical/tube thoracostomy in 26 (65 %) patients, and fistula (bronchopleural, esophagopleural and biliopleural) in 12 (30 %) patients. The 14-day and 30-day mortality rates were 32.5 % and 42.5 %, respectively. Pleural infections due to S. maltophilia are most commonly the result of surgical procedures, thoracostomy, and underlying fistulas. These infections are associated with a high mortality rate, especially among immunocompromised patients.

Effect of the beta secretase-1 inhibitor on the amyloid C-terminal fragment of amyloid precursor protein processing in a hyperphosphorylated tau rat model.

The amyloid C-terminal fragment (ßCTF) of the amyloid precursor protein (APP) is the cleaved component of APP by beta secretase-1 (BACE1), which shows similar neurotoxicity as amyloid beta (Aß) in many ways. Evidence suggested that in addition to Aß, ßCTF might also participate in the pathogenesis of Alzheimer's disease (AD). In recent years, the relationship between ßCTF processing and hyperphosphorylated tau has attracted increasing research attention. In this study, we established an animal model of tau hyperphosphorylation with okadaic acid (OA) treatment, and analyzed ßCTF processing in vivo. The ßCTF level was found to increase in neurons, which was most likely caused by the induction of OA and BACE1 overexpression. Furthermore, these results provide the first evidence that ßCTF can predominately accumulate in the axons of neurons in a hyperphosphorylated tau state in vivo, and suggested that the redistribution of ßCTF is involved in the pathogenesis of AD. These results indicate that BACE1 could be a therapeutic target of AD by affecting the processing of ßCTF.

Isolated familial pneumothorax in a Taiwanese family with Birt-Hogg-Dubé syndrome.

Primary spontaneous pneumothorax usually occurs as a sporadic event, but may be clustered in certain families with an underlying inherited disorder. Birt-Hogg-Dubι (BHD) syndrome is a rare autosomal dominant disease accounting for familial pneumothorax. BHD syndrome, caused by mutation of the folliculin gene, is characterized by skin fibrofolliculoma, pulmonary cysts, pneumothorax, and renal cancer. We describe a BHD-affected Taiwanese family with clinical and genetic study. A rare mutation of the folliculin gene was detected in the patient and members with pulmonary cysts or pneumothorax, but no skin or renal lesions were found. This mutation was reported in a Taiwanese family and might indicate a pneumothorax-predominant phenotype. Isolated pneumothorax is an uncommon initial presentation of BHD syndrome. Family history should be carefully reviewed when managing a patient with pneumothorax.

CRISPR applications for Duchenne muscular dystrophy: From animal models to potential therapies.

CRISPR gene-editing technology creates precise and permanent modifications to DNA. It has significantly advanced our ability to generate animal disease models for use in biomedical research and also has potential to revolutionize the treatment of genetic disorders. Duchenne muscular dystrophy (DMD) is a monogenic muscle-wasting disease that could potentially benefit from the development of CRISPR therapy. It is commonly associated with mutations that disrupt the reading frame of the DMD gene that encodes dystrophin, an essential scaffolding protein that stabilizes striated muscles and protects them from contractile-induced damage. CRISPR enables the rapid generation of various animal models harboring mutations that closely simulates the wide variety of mutations observed in DMD patients. These models provide a platform for the testing of sequence-specific interventions like CRISPR therapy that aim to reframe or skip DMD mutations to restore functional dystrophin expression. This article is categorized under: Congenital Diseases > Genetics/Genomics/Epigenetics.

RNA is favourable for analysing EGFR mutations in malignant pleural effusion of lung cancer.

Malignant pleural effusion (MPE) is a useful specimen allowing for the evaluation of EGFR status in nonsmall cell lung cancer (NSCLC). However, direct sequencing of genomic DNA from MPE samples was found not to be sensitive for EGFR mutation detection. To test whether EGFR analysis from RNA is less prone to interference from nontumour cells that have no or lower EGFR expression, we compared three methods (sequencing from cell-derived RNA versus sequencing and mass-spectrometric analysis from genomic DNA), in parallel, for EGFR mutation detection from MPE samples in 150 lung adenocarcinoma patients receiving first-line tyrosine kinase inhibitors (TKIs). Among these MPE samples, EGFR mutations were much more frequently identified by sequencing using RNA than by sequencing and mass-spectrometric analysis from genomic DNA (for all mutations, 67.3 versus 44.7 and 46.7%; for L858R or exon 19 deletions, 61.3 versus 41.3 and 46.7%, respectively). The better mutation detection yield of sequencing from RNA was coupled with the superior prediction of clinical efficacy of first-line TKIs. In patients with acquired resistance, EGFR sequencing from RNA provided satisfactory detection of T790M (54.2%). These results demonstrated that EGFR sequencing using RNA as template greatly improves sensitivity for EGFR mutation detection from samples of MPE, highlighting RNA as the favourable source for analysing EGFR mutations from heterogeneous MPE specimens in NSCLC.

First-line nivolumab + ipilimumab in advanced NSCLC: CheckMate 227 subpopulation analyses in Asian patients.

BACKGROUND: Nivolumab plus ipilimumab demonstrated clinically meaningful improvement in efficacy versus chemotherapy with a manageable safety profile in patients with advanced non-small cell lung cancer (NSCLC) and tumor programmed death-ligand 1 (PD-L1) expression ≥1% or <1% in Part 1 of CheckMate 227. Here we report efficacy and safety results for the Asian subpopulation. METHODS: Patients with stage IV/recurrent NSCLC were randomized 1 : 1 : 1 to nivolumab plus ipilimumab, nivolumab monotherapy, or chemotherapy (PD-L1 ≥1%) or nivolumab plus ipilimumab, nivolumab plus chemotherapy, or chemotherapy (PD-L1 <1%). Overall survival (OS), progression-free survival, objective response rate, duration of response, and safety were evaluated among patients in Japan, South Korea, and Taiwan. RESULTS: In the Asian subpopulation with PD-L1 ≥1%, 81 patients received nivolumab plus ipilimumab and 81 received chemotherapy. Median OS was not reached with nivolumab plus ipilimumab versus 24.8 months with chemotherapy; 3-year OS rate was 53% versus 37% [hazard ratio (HR), 0.72; 95% confidence interval (CI) 0.47-1.11]. The 3-year progression-free survival rate was 26% versus 7% (HR, 0.65; 95% CI 0.45-0.96), objective response rate was 56% versus 37%, and median duration of response was 29.0 months (95% CI 15.0 months-not reached) versus 6.9 months (95% CI 3.9-11.1 months). Similar results were observed regardless of tumor PD-L1 expression and in Japanese patients. Grade 3-4 treatment-related adverse events occurred in 40% of patients receiving nivolumab plus ipilimumab and 36% receiving chemotherapy, in the overall Asian subpopulation (tumor PD-L1 expression ≥1% and <1%); no new safety signals were identified. CONCLUSIONS: At 3-year follow-up, nivolumab plus ipilimumab provided durable long-term efficacy benefits versus chemotherapy regardless of tumor PD-L1 expression in the Asian subpopulation, including Japanese patients. Consistent with findings for all randomized patients, these data support the use of nivolumab plus ipilimumab as first-line treatment of Asian patients with advanced NSCLC.

Diagnostic value of procalcitonin in pleural effusions.

This study was to determine the diagnostic value of procalcitonin (PCT) in the differentiation of infectious and non-infectious causes of pleural effusion. From January 2005 to April 2005, we measured the PCT levels of pleural effusion from 76 patients using an immunoluminometric assay. The types of pleural infusions studied were para-pneumonic effusion (n = 26), empyema (n = 7), tuberculous pleurisy (n = 8), malignant pleural effusion (n = 25) and transudative pleural effusion (n = 8). The PCT levels were low in transudative pleural effusions (0.188 ± 0.077 ng/mL) and tuberculous pleurisy (0.130 ± 0.069 ng/mL), but high in empyema (5.147 ± 3.056 ng/mL), para-pneumonic effusion (1.091 ± 0.355 ng/mL), and malignant pleural effusion (0.241 ± 0.071 ng/mL). The receiver-operating characteristic curve analysis for an optimal discrimination between empyema and para-pneumonic effusion from non-para-pneumonic effusion could be performed at a cut-off point of 0.18 ng/mL with area under the curve of 0.776 (sensitivity: 69.7%, specificity: 72.1%). The correlation was found between pleural effusion PCT and serum PCT levels in 16 patients (r² = 0.967, p < 0.001). In conclusion, a high pleural effusion PCT level suggests the presence of empyema and para-pneumonic effusion.

Bacteremia caused by Brevundimonas species at a tertiary care hospital in Taiwan, 2000-2010.

We investigated clinical and microbiological characteristics of 30 patients with Brevundimonas bacteremia treated at a tertiary care hospital in Taiwan during 2000-2010. All the 30 bacteria isolates were confirmed to the species level by 16S rRNA sequencing analysis. Minimum inhibitory concentrations (MICs) of 11 antimicrobial agents against these isolates were determined by the agar dilution method. Seventeen (57%) patients had underlying malignancy, 12 (40%) had undergone central catheter placement, and 13 (43%) had received chemotherapy within the previous three months. Eight (27%) patients had community-acquired bacteremia and the remaining 22 patients (73%) had healthcare-associated bacteremia. The overall 14-day and 30-day mortality rates were 13% and 17%, respectively. Among the 30 isolates, B. vesicularis constituted most commonly (n = 22, 63%), followed by B. nasdae (n = 5) and B. diminuta (n = 3). All isolates were susceptible to piperacillin-tazobactam and amikacin, while all were resistant to ciprofloxacin and colistin. Tigecycline (MICs at which 90% of isolates are inhibited [MIC(90)] was 0.12 mg/L) and doripenem (MIC(90) of 1 mg/L) both possessed good in vitro activities. In conclusions, Brevundimonas should be considered a pathogen that can cause bacteremia in immunocompromised hosts. Piperacillin-tazobactam, amikacin, doripenem, and tigecycline exhibit good in vitro activities against these ciprofloxacin- and colistin-resistant Brevundimonas species.

Skin and soft-tissue infection caused by non-tuberculous mycobacteria in Taiwan, 1997-2008.

The aim of this study was to investigate the clinical, microbiological, and pathological characteristics and the outcomes of skin and soft-tissue infection (SSTI) caused by non-tuberculous mycobacteria (NTM). Medical records of 50 patients with SSTI caused by NTM identified from 2005 to 2008 and 63 patients previously reported in a medical centre from 1997 to 2004 were reviewed. The annual incidence (per 100,000 outpatients and in-patients) ranged from 0·57 in 2005, 0·38 in 2007, to 1·1 in 2008, with an average of 0·62/100,000. From 1997 to 2008, the average incidence was 1·39/100,000 patients. The average annual incidence of SSTI caused by NTM was 0·62/100,000 outpatients and in-patients during 2005 and 2008. Of the total of 113 patients identified during the 12-year period, patients infected with Mycobacterium fortuitum and M. marinum were younger than those infected with M. avium-intracellulare complex (MAC) (36 and 44 years vs. 55 years, P=0·004 and P=0·056, respectively), and were more likely to have previous invasive procedures than those infected with MAC and M. abscessus (81·8% and 72·0% vs. 27·8% and 54·8%, P=0·007), and less likely to have associated immunosuppression (9·1% and 24% vs. 66·7% and 45·2%, P=0·006). Granuloma was more often observed in immunocompetent patients (60·1% vs. 40%, P=0·019), and in M. marinum-infected specimens (78·3%). There were significant differences in the demographic and clinical features of patients with NTM SSTI, including immunosuppression, trauma experience, and depth of tissue infections.

Imaging patterns and prognosis of patients with gefitinib-related interstitial lung disease.

PURPOSE: We aimed to summarize the imaging findings of 25 patients with gefitinib-related interstitial lung disease (ILD), and identify the factors related to prognosis of gefitinib-related ILD in patients with non-small-cell-lung cancer. MATERIALS AND METHODS: Diagnosis of gefitinib-induced ILD by at least two chest radiologists was based on a review and analysis of the chest radiography and CT findings plus clinical data in the medical records. All patients were diagnosed with Stage III - IV non-small-cell carcinoma (adenocarcinoma (n = 24), bronchioalveolar cell carcinoma (n = 1)) and essential clinical data such as gefitinib as first-line use and survival status were recorded and analyzed to determine whether these were prognosis predictors. The imaging findings were classified into four patterns according to the previous largest study in Japan. RESULTS: The 25 chest radiographs were classified as Pattern A (n = 8), Pattern B (n = 3), Pattern C (n = 6), and pattern D (n = 8). Likewise the 23 CT images were classified as pattern A (n = 8; 34.8%), B (n = 3; 13%), C (n = 5; 21.7%), and D (n = 7; 30.4%). The mortality rate was significantly higher in patients with pattern D than in patients with the other patterns. Pattern D imaging findings were also significantly correlated with non first-line use of gefitinib (p = 0.007). CONCLUSIONS: We found an increase in mortality rate in patients with gefitinib associated ILD/pattern D compared to other radiological patterns. Familiarity with these imaging patterns can facilitate early and accurate diagnosis and help physicians gauge clinical prognosis of gefitinib-related ILD.

The diagnosis of lung cancer in the era of interventional pulmonology.

Advances in bronchoscopic and other interventional pulmonology technologies have expanded the sampling procedures pulmonologist can use to diagnose lung cancer and accurately stage the mediastinum. Among the modalities available to the interventional pulmonologist are endobronchial ultrasound-guided transbronchial needles aspiration (EBUS-TBNA) and transoesophageal bronchoscopic ultrasound-guided fine-needle aspiration (EUS-B-FNA) for sampling peribronchial/perioesophageal central lesions and for mediastinal lymph node staging, as well as navigational bronchoscopy and radial probe endobronchial ultrasound (RP-EBUS) for the diagnosis of peripheral lung cancer. The role of the interventional pulmonologist in this setting is to apply these procedures based on the correct interpretation of clinical and radiological findings in order to maximise the chances of achieving the diagnosis and obtaining sufficient tissue for molecular biomarker testing to guide targeted therapies for advanced non-small cell lung cancer. The safest and the highest diagnosis-yielding modality should be chosen to avoid a repeat sampling procedure if the first one is non-diagnostic. The choice of site and biopsy modality are influenced by tumour location, patient comorbidities, availability of equipment and local expertise. This review provides a concise state-of-the art account of the interventional pulmonology procedures in the diagnosis and staging of lung cancer.