Highly efficient CRISPR-mediated genome editing through microfluidic droplet cell mechanoporation.

Clustered regularly interspaced short palindromic repeats (CRISPR)-based editing tools have transformed the landscape of genome editing. However, the absence of a robust and safe CRISPR delivery method continues to limit its potential for therapeutic applications. Despite the emergence of various methodologies aimed at addressing this challenge, issues regarding efficiency and editing operations persist. We introduce a microfluidic gene delivery system, called droplet cell pincher (DCP), designed for highly efficient and safe genome editing. This approach combines droplet microfluidics with cell mechanoporation, enabling encapsulation and controlled passage of cells and CRISPR systems through a microscale constriction. Discontinuities created in cell and nuclear membranes upon passage facilitate the rapid CRISPR-system internalization into the nucleus. We demonstrate the successful delivery of various macromolecules, including mRNAs (~98%) and plasmid DNAs (~91%), using this platform, underscoring the versatility of the DCP and leveraging it to achieve successful genome engineering through CRISPR-Cas9 delivery. Our platform outperforms electroporation, the current state-of-the-art method, in three key areas: single knockouts (~6.5-fold), double knockouts (~3.8-fold), and knock-ins (~3.8-fold). These results highlight the potential of our platform as a next-generation tool for CRISPR engineering, with implications for clinical and biological cell-based research.

Effect of Age at Cochlear Implantation in Educational Placement and Peer Relationships.

OBJECTIVES: The education and school life of children who have undergone cochlear implantation (CI) is very important and should be monitored continuously. This study assessed auditory performance in children with cochlear implants over time, along with educational placement and peer relationship, and compared the results based on the age at CI and the presence of additional disabilities. DESIGN: In total, 77 children who had undergone CI at the Hearing Center in Ajou University Hospital at less than 10 years old and who were presently attending school or had already graduated from school within 3 years were enrolled in this study. All children had congenital bilateral severe or profound hearing loss at the diagnosis. They were classified based on the age at which they received CI: the "early-implanted group," younger than 3.5 years (n = 38), and the "late-implanted group," between 3.5 and 10 years old (n = 39). RESULTS: The early-implanted group had worse short-term auditory performance than the late group, but after 2 years of the implant use, auditory performance was similar in both groups. The early and late groups did not differ significantly in terms of the proportions of children who were enrolled in a regular school (94.7% and 89.7%, respectively). However, the early-implanted group had a larger proportion of children who were enrolled in a regular school without additional needs (73.0%), compared with the late group (48.6%) (p = .034). Children with multiple disabilities showed poorer performance (word score of 57.7% and sentence score of 44.7%) than children with hearing disability only (91.8% and 87.2%, respectively), which affected full-time enrollment in regular schools. With regard to peer relationships, 19.0% of children in the early-implanted group required close observation and assistance, and 9.5% even required help and counseling. Children who underwent early CI had a high tendency toward social restraint, apathy, and over-commitment. No correlations were observed between audiological factors and the aspects of peer relationships assessed in this study. CONCLUSIONS: Early CI and the absence of other disabilities were the two main factors that increased the likelihood of full-time enrollment in mainstream classes at regular schools. Nevertheless, many children who underwent earlier CI still encountered difficulties in peer relationships.

Antioxidant, Antimicrobial and Anti-Inflammatory Activities of Essential Oil Derived from the Wild Rhizome of Atractylodes macrocephala.

The present study investigated the chemical composition, antioxidant, antimicrobial, and anti-inflammatory activities of essential oil (EO) derived from the wild rhizomes of Atractylodes macrocephala Koidz. (AMA) growing in Qimen County (eastern China). GC/MS analysis identified fifteen compounds, representing 92.55 % of AMA EO. The major compounds were atractylone (39.22 %), ß-eudesmol (27.70 %), thymol (5.74 %), hinesol (5.50 %), and 11-isopropylidenetricyclo[4.3.1.1(2,5)]undec-3-en-10-one (4.71 %). Ferricyanide reducing, 1,1-diphenyl-2-picyrlhydrazyl (DPPH) and 3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) scavenging assays revealed that AMA EO exhibited strong antioxidant capacities. Additionally, AMA EO showed inhibitory effects on growth of Escherichia coli, Pseudomonas aeruginosa, Salmonella enterica, Staphylococcus aureus, and Bacillus subtilis, with the minimum inhibitory concentrations (MIC) ranging from 0.5 to 2.0 mg/mL. Treatments with AMA EO also significantly inhibited nitric oxide (NO) and prostaglandin E2 (PGE2 ) production in lipopolysaccharide-stimulated RAW264.7 cells, indicating anti-inflammatory activity of AMA EO. Furthermore, treatments with AMA EO decreased the transcriptional levels of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2), which might be the molecular mechanisms underlying its anti-inflammatory effects. Overall, these results provide a theoretical basis for further study and application of AMA EO in food and medicine products.

Anti-inflammatory effects of mulberry (Morus alba L.) root bark and its active compounds.

Mulberry (Morus alba L.) root bark (MRB) was extracted using methanol and the extracts were subjected to tests of anti-inflammatory effects. The ethyl acetate fraction demonstrated the best anti-inflammatory effects. Purified compounds, sanggenon B, albanol B and sanggenon D, showed inhibitory effects on NO production in LPS-stimulated RAW264.7 cells and albanol B demonstrated the best anti-inflammatory effects. Regarding the underlying molecular mechanisms, further investigations showed that treatments with Albanol B reduced production of pro-inflammatory cytokines and decreased expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2). These results would contribute to development of novel anti-inflammatory drugs from MRB.

Critical Factors for Binaural Hearing in Children with Bilateral Sequential Cochlear Implantation: First Implant Performance and Inter-Implant Interval.

OBJECTIVES: This study evaluated the influence of the performance of the first cochlear implant (CI-1) on the second implant (CI-2) and the significance of inter-implant intervals in children with sequential cochlear implantation. METHOD: Thirty-four patients were included for speech perception outcome and 38 patients were included for subjective hearing satisfaction in daily life in this study. The patients were classified into 3 groups according to inter-implant interval: group I, <4 years; group II, 4-7 years; group III, >7 years. Open-set speech perception scores before and at 3, 6, 12, and 24 months after the second implantation was compared among the CI-1, CI-2, and bilateral CI conditions. Subjective hearing benefits in daily life were measured using a questionnaire of Speech, Spatial, and Qualities of Hearing Scale (SSQ) for parents of children with impaired hearing, and device use and preference were evaluated (by phone interview.) Results: The speech perception of children using the CI-2 only improved quickly within 3 months postoperatively and were similar to those using the CI-1 only. The results of monosyllabic and disyllabic word tests using CI-2 at 3 months after second implantation were not significantly different from those in patients using the CI-1 only. In the sentence test, the scores using the CI-2 only were not significantly different from those using the first implant only at 6 months after second implantation. SSQ scores were similar among groups and the worst score in each section was shown from the questions about performance under noisy conditions in each section. Device usage habits showed that 17 (44.7%) and 16 (42.1%) patients preferred bilateral and the CI-1, respectively. Only 5 (13.2%) patients wanted the CI-2 rather than the CI-1. While group I and II showed a prominent positive attitude to bilateral use, group III, for which the inter-implant interval was 7 years or more, showed a definite preference for the CI-1 (p < 0.01). CONCLUSIONS: Functional binaural benefits were achieved in patients who were good performers with the CI-1 after the second implantation irrespective of the inter-implant interval. Sequential CI should be strongly recommended for patients with unilateral CI showing a good performance.

Recurrence Rate of Herpes Zoster and Its Risk Factors: a Population-based Cohort Study.

BACKGROUND: Herpes zoster (HZ) is generally thought to occur once in a lifetime and recurrence is considered to be limited to immunocompromised individuals. Although HZ recurrence rates seem to be increasing, there have been few studies exploring these rates in the general population. We investigated the recurrence rate and associated risk factors in the general population. METHODS: We used the population-based samples of the National Health Insurance Service database to identify cases of initial HZ episodes from January 1, 2002 to December 31, 2013. We also followed up on these cases through December 31, 2013 to identify recurrence. RESULTS: Overall, the incidence rate of HZ is 5.1 per 1,000 person years and the recurrence rate is 12.0 per 1,000 person years. There were 2,100 recurrent cases out of 39,441 initial episodes with 4.4 years of the mean follow-up period. We identified significant risk factors for recurrence such as old age (51-70 years) (hazard ratio [HR], 1.447; 95% confidence interval [CI], 1.311-1.598), women (1.476; 1.345-1.619), zoster-related pain (ZRP) longer than 30 days (cases of ZRP lasting 31-90 days [1.200; 1.042-1.383], and ZRP lasting longer than 90 days [2.293; 1.990-2.643]). Concurrent hematologic malignancies (2.864; 1.929-4.251), autoimmune diseases (1.466; 1.252-1.715), dyslipidemia (1.390; 1.263-1.530), and hypertension (1.222; 1.107-1.350) were also significant risk factors. CONCLUSION: Our results suggest that the recurrence of HZ is much more common than generally expected, and that the associated risk factors can play an important role in predicting recurrence.

Repeated Glucose Deprivation/Reperfusion Induced PC-12 Cell Death through the Involvement of FOXO Transcription Factor.

BACKGROUND: Cognitive impairment and brain damage in diabetes is suggested to be associated with hypoglycemia. The mechanisms of hypoglycemia-induced neural death and apoptosis are not clear and reperfusion injury may be involved. Recent studies show that glucose deprivation/reperfusion induced more neuronal cell death than glucose deprivation itself. The forkhead box O (FOXO) transcription factors are implicated in the regulation of cell apoptosis and survival, but their role in neuronal cells remains unclear. We examined the role of FOXO transcription factors and the involvement of the phosphatidylinositol 3-kinase (PI3K)/Akt and apoptosis-related signaling pathways in PC-12 cells exposed to repeated glucose deprivation/reperfusion. METHODS: PC-12 cells were exposed to control (Dulbecco's Modified Eagle Medium [DMEM] containing 25 mM glucose) or glucose deprivation/reperfusion (DMEM with 0 mM glucose for 6 hours and then DMEM with 25 mM glucose for 18 hours) for 5 days. MTT assay and Western blot analysis were performed for cell viability, apoptosis, and the expression of survival signaling pathways. FOXO3/4',6-diamidino-2-phenylindole staining was done to ascertain the involvement of FOXO transcription factors in glucose deprivation/reperfusion conditions. RESULTS: Compared to PC-12 cells not exposed to hypoglycemia, cells exposed to glucose deprivation/reperfusion showed a reduction of cell viability, decreased expression of phosphorylated Akt and Bcl-2, and an increase of cleaved caspase-3 expression. Of note, FOXO3 protein was localized in the nuclei of glucose deprivation/reperfusion cells but not in the control cells. CONCLUSION: Repeated glucose deprivation/reperfusion caused the neuronal cell death. Activated FOXO3 via the PI3K/Akt pathway in repeated glucose deprivation/reperfusion was involved in genes related to apoptosis.

Ginsenoside Rg3 prevents INS-1 cell death from intermittent high glucose stress.

BACKGROUND: Ginsenoside Rg3 has been proposed to mediate anti-diabetic effects, but their direct effect on pancreatic ß cell viability and mechanisms are not clearly understood. Recent studies suggest that intermittent high glucose (IHG) could be more harmful to pancreatic ß cells than sustained high glucose. There are few reports about the effect of the ginsenosideRg3 to ß cell apoptosis and proliferation against IHG. METHODS: INS-1 cells were treated with alternative glucose concentration with or without ginsenoside Rg3. Cell apoptosis and viability were detected by Annexin V staining and MTT assay. The activation of mitogen-activated protein kinases (MAPKs) was analyzed by Western blotting using specific antibodies. Quantification of secreted insulin protein was measured using rat/mouse Insulin ELISA kits. Bromodeoxyuridine (BrdU) staining and florescence in situ hybridization (FISH) analysis was performed to compare cell proliferation. RESULT: INS-1 cell viability was decreased under IHG and increased with Rg3 treatment.Rg3 significantly reduced the apoptotic INS-1 cells against IHG. The quantification of secreted insulin concentration was increased with Rg3. Rg3 increased INS-1 cell proliferation. ERK and p38 MAPK pathways reduced by IHG were activated by the ginsenoside Rg3. CONCLUSION: Ginsenoside Rg3 protected INS-1 cell death from IHG with reducing apoptosis and increasing proliferation.

Co-Culture of α TC-6 Cells and ß TC-1 Cells: Morphology and Function.

BACKGROUND: In vitro experiments using only ß-cell lines instead of islets are limited because pancreatic islets are composed of four different types of endocrine cells. Several recent studies have focused on cellular interactions among these cell types, especially α- and ß-cells. Because islet isolation needs time and experience, we tested a simple co-culture system with α- and ß-cells. Their morphology and function were assessed by comparison to each single cell culture and pancreatic islets. METHODS: α TC-6 cells and ß TC-1 cells were maintained in Dulbecco's Minimal Essential Medium containing 5 mM glucose and 10% fetal bovine serum. Cells were mixed at a 1:1 ratio (5×105) in 6-well plates and cultured for 24, 48, and 72 hours. After culture, cells were used for insulin and glucagon immunoassays and tested for glucose-stimulated insulin secretion (GSIS). RESULTS: α TC-6 and ß TC-1 cells became condensed by 24 hours and were more strongly compacted after 48 hours. ß TC-1 cells showed both ß-ß and ß-α cell contacts. GSIS increased with increasing glucose concentration in co-cultured cells, which showed lower secreted insulin levels than ß TC-1 cells alone. The increase in the secreted insulin/insulin content ratio was significantly lower for co-cultured cells than for ß-cells alone (P=0.04). Compared to islets, the α-/ß-cell co-culture showed a higher ratio of GSIS to insulin content, but the difference was not statistically significant (P=0.09). CONCLUSION: α TC-6 and ß TC-1 cells in the co-culture system showed cell-to-cell contacts and a similar stimulated insulin secretion pattern to islets. The co-culture system may be used to better mimic pancreatic islets in in vitro assessments.

Diabetic myonecrosis in a patient with hepatitis B-induced liver cirrhosis.

Diabetic myonecrosis-a rare complication of long-standing, poorly controlled diabetes mellitus-typically presents with acute-onset muscle pain, is self-limiting, and responds well to conservative management. We report a case of diabetic myonecrosis in a 33-year-old man with hepatitis B-induced liver cirrhosis and type 2 diabetes who presented with abdominal distension and pain in the left thigh. Diabetic myonecrosis was diagnosed based on clinical presentation, radiological findings, magnetic resonance imaging and histopathological investigations; he was successfully treated conservatively with insulin and analgesics. Diabetic myonecrosis should be considered in the differential diagnosis of muscle pain in patients with diabetes.

The effect of proton pump inhibitors on glycated hemoglobin levels in patients with type 2 diabetes mellitus.

OBJECTIVE: Gastrin increases the growth and neogenesis of the islets of Langerhans. Oral proton pump inhibitors (PPIs) increase the circulating gastrin level in animals and humans, but the therapeutic benefit of PPIs for diabetes mellitus has not been resolved. We examined whether treatment with a PPI for ≥2 months affected the glycated hemoglobin (A1C) level in patients with type 2 diabetes. METHODS: The electronic medical records of patients treated at the Busan Paik Hospital in South Korea were examined. The primary outcome measure was the change in A1C before and after PPI treatment for ≥2 months. We also tested if the primary outcome measure was affected by sex, age, duration of diabetes, body mass index, PPI molecule, duration of treatment with PPI or concurrent therapy with other antidiabetes agents. RESULTS: In total, 43 patients (17 men and 26 women) were studied (mean age 63.8 years). Patients were treated with a PPI for a mean of 180 days. The A1C levels before and after treatment were not significantly different (6.86%±1.10% and 6.77%±1.07%, respectively; p=0.406). Metformin monotherapy did not lower A1C levels as compared with a combination therapy including metformin and antidiabetes medication not including metformin. CONCLUSIONS: Proton pump inhibitor treatment of patients with type 2 diabetes did not reduce A1C levels. The data of this study were obtained from a retrospective chart review and included a small number of subjects. Furthermore, large randomized controlled studies are needed to define the effect of PPIs for type 2 patients with diabetes.

Population-based study of the epidemiology of herpes zoster in Korea.

General epidemiological data regarding herpes zoster (HZ) are necessary for treatment and prevention of this disease. In addition, epidemiological data can play an important role in evaluating the efficacy and impact of vaccination. Though several epidemiological studies of HZ in Korea have been conducted, they usually depend on hospital-based data and may not be representative of HZ characteristics all over Korea. The purpose of this study was to evaluate the incidence and other epidemiological features of HZ in the general Korean population. We used population-based medical records from the Health Insurance Review & Assessment Service, which includes 50,908,646 medical insurance subscribers, to calculate the incidence of HZ. Also, we analyzed an age-stratified random sample of 1,375,842 individuals to study descriptive epidemiologic characteristics of HZ in Korea in 2011. We observed that the incidence of HZ was 10.4 per 1,000 person-years and was strongly correlated with age. Sex had a major influence on HZ incidence; overall, there were 12.6 cases per 1,000 person years in women and 8.3 cases per 1,000 person years in men. There was no difference in incidence according to the locality and season.

Association of low muscle mass and combined low muscle mass and visceral obesity with low cardiorespiratory fitness.

OBJECTIVE: Previous studies have shown that low cardiorespiratory fitness (CRF), visceral obesity and low muscle mass may share pathophysiological mechanisms, such as insulin resistance and chronic inflammation. In this study, we investigated whether low CRF is associated with low muscle mass, visceral obesity, and visceral obesity combined with low muscle mass. RESEARCH DESIGN AND METHODS: The associations between CRF and low muscle mass and combined low muscle mass and visceral obesity were examined in 298 apparently healthy adults aged 20-70 years. Low muscle mass was defined using a skeletal muscle mass index (SMI) that was calculated using dual energy X-ray absorptiometry. Visceral obesity was defined as a visceral fat area (VFA) exceeding 100 cm2 in women and 130 cm2 in men. We classified the participants into 4 low muscle mass/visceral obesity groups according to SMI and VFA. CRF was measured using a cycle ergometer test. RESULTS: CRF level correlated positively with SMI and negatively with VFA. Individuals with low muscle mass had lower CRF values than those without low muscle mass. After adjustment for age, sex, lifestyle factors, and markers for insulin resistance and inflammation, participants in the lowest quartile of CRF had an odds ratio (OR) for low muscle mass of 4.98 compared with those in the highest quartile (95% confidence interval (CI) = 1.19-12.99; P for trend = 0.001) and an OR for combined low muscle mass and visceral obesity of 31.46 (95% CI = 4.31-229.68; P for trend = 0.001). CONCLUSIONS: Individuals with lower CRF exhibited increased risk of low muscle mass and combined low muscle mass and visceral obesity. These results suggest that low CRF may be a potential indicator for low muscle mass and combined low muscle mass and visceral obesity in Korean adults.

Pregnancy in postpartum estrus induces inflammatory milk production and catagen specific pup skin inflammation in interleukin-10 deficient mice.

BACKGROUND: The interleukin 10 deficient mice (IL-10(-/-)) showed high incidence of pup alopecia compared to other strains, and pup alopecia was caused by skin inflammation and was recoverable. Pup alopecia of B6.IL-10(-/-) might be related with maternal factor and interleukin-10 deficient phenotype. OBJECTIVE: The objectives of this study were elucidating of maternal factors for inflammatory milk production and characterization of pup alopecia in IL-10(-/-) mice. METHODS: Incidences of pup alopecia were analyzed with 13 breeding cases. Comparison between control and alopecia pups and its dams, were conducted with histological examination (H&E, TUNEL assay, immunohistochemistry for F4/80, iNOS, CD206, Gr-1, CD4, CD8, CD11c and CD326), fostering test, forced weaning test, qPCR for tyrosine hydroxylase, flow cytometry, IL-10 inhibition test, BMDM stimulation test and LC/MS analysis. RESULTS: Presence of pregnancy in postpartum estrus showed significant correlation with inflammatory milk production and mammary gland involution in B6.IL-10(-/-) mice. There were no different mass in inflammatory milk, but different ionization intensity was detected. Inflammatory milk directly induced hepatocyte steatosis, catagen stage specific hair breaking and alopeicia in pups. Histologically, hypertropy of outer root sheath and macrophage/neutrophil infiltration were typical. CONCLUSION: B6.IL-10(-/-) dam with stress such as PPE could produce untimely mammary gland involution and inflammatory milk production. Interleukin 10 is important for maternal stress regulation and protecting inflammatory milk production, also influence severity of pup skin inflammation and alopecia. Remarkably, inflammatory milk induced hepatocyte steatosis, and it could indicate there is abnormal lipid metabolism. This was first report for catagen specific alopecia in mouse.

Serum adipocyte fatty acid-binding protein levels are independently associated with sarcopenic obesity.

AIMS: Adipocyte fatty acid-binding protein (A-FABP) plays a key role in obesity-related insulin resistance and inflammation which may be involved in the pathogenesis of sarcopenic obesity (SO). This study examined the association of SO with serum A-FABP levels in Korean adults. METHODS: Two hundred ninety eight adults aged 20-70 years were examined using dual X-ray absorptiometry and computed tomography and measuring serum A-FABP levels. Sarcopenia was defined as the appendicular skeletal muscle mass (ASM) divided by weight (%) of <1 SD below the mean values of young adults in both sexes. Obesity was defined as visceral fat area (VFA) ≥ 100 cm(2). RESULTS: Serum A-FABP levels were higher in groups with SO compared to non-SO groups in both men and women. In the unadjusted model, serum A-FABP levels were positively associated with VFA and negatively associated with ASM/weight. Even after adjusting for possible confounding factors, ASM/weight was found to be independently and negatively associated with serum A-FABP levels. In addition, multiple logistic regression analysis showed that increased serum levels of A-FABP were independently associated with the presence of SO. CONCLUSIONS: The present findings indicate that serum A-FABP levels may be valuable markers of the presence of SO.

A1c variability can predict coronary artery disease in patients with type 2 diabetes with mean a1c levels greater than 7.

BACKGROUND: Recent studies suggested that the association of acute glucose variability and diabetic complications was not consistent, and that A1c variability representing long term glucose fluctuation may be related to coronary atherosclerosis in patients with type 1 diabetes. In this study, we attempt to determine whether or not A1c variability can predict coronary artery disease (CAD) in patients with type 2 diabetes. METHODS: We reviewed data of patients with type 2 diabetes who had undergone coronary angiography (CAG) and had been followed up with for 5 years. The intrapersonal standard deviation (SD) of serially-measured A1c levels adjusted by the different number of assessments among patients (adj-A1c-SD) was considered to be a measure of the variability of A1c. RESULTS: Among the 269 patients, 121 of them had type 2 diabetes with CAD. In patients with A1c ≥7%, the mean A1c levels and A1c levels at the time of CAG among the three groups were significantly different. The ratio of patients with CAD was the highest in the high adj-A1c-SD group and the lowest in the low adj-A1c-SD group (P=0.017). In multiple regression analysis, adj-A1c-SD was an independent predictor for CAD in subjects with A1c ≥7% (odds ratio, 2.140; P=0.036). CONCLUSION: Patients with higher A1c variability for several years showed higher mean A1c levels. A1c variability can be an independent predictor for CAD as seen in angiographs of patients with type 2 diabetes with mean A1c levels over 7%.

A simple separation method with a microfluidic channel based on alternating current potential modulation.

A simple separation and detection system based on an electrochemical potential modulated microchannel (EPMM) device was developed for the first time. The application of alternating current (AC) potential to the microfluidic separation channel walls, which were composed of screen printed carbon electrodes, resulted in the oscillation and fluctuation of analytes and in the formation of a perfect flat flow front. These events resulted in an increase in the effective concentration and in the fine separation of samples. The performance of the EPMM device was examined through the analysis of endocrine disruptors (EDs) and heavy metal ions (HMIs) as model compounds. The analytical parameters that affected the separation and detection of EDs and HMIs were studied in terms of AC amplitude, AC frequency, flow rate, buffer concentration, pH, detection potential, and temperature. The separation efficiency was evaluated through measurements of the theoretical plate number (N), the retention time, and the half-peak width. Linear calibration plots for the detection of EDs and HMIs were obtained between 0.15 and 250.0 nM (detection limit 86.4 ± 2.9 pM) and between 0.01 and 10.0 nM (detection limit 9.5 ± 0.3 pM), respectively. The new device was successfully demonstrated with authentic and real samples.

Perforated Mitral Valve Aneurysm in the Posterior Leaflet without Infective Endocarditis.

Aneurysm of the mitral valve, although uncommon, occurs most commonly in association with infective endocarditis of the aortic valve and true mitral valve aneurysm is a rare cause of mitral regurgitation. We report a case with perforated mitral valve aneurysm in the posterior leaflet without concurrent infective endocarditis initially mistaken diagnosis of cystic mass, which was confirmed at operation with successful mitral valve annuloplasty.

Effects of Q-switched and long-pulsed 1064 nm Nd:YAG laser on enlarged facial pores.

'Enlarged facial pore' is a subjective term, which is not clearly defined but often complained by many. A diverse range of treatments are used though evidence of efficacy remains largely anecdotal. We report a series of nine patients who underwent a split face trial with Q-switched 1064 nm Nd:YAG and long-pulsed 1064 nm Nd:YAG laser to treat enlarged facial pores.