The hiPSC-derived corneal endothelial progenitor-like cell recovers the rabbit model of corneal endothelial dystrophy.

INTRODUCTION: Corneal endothelial dysfunction results in cornea opacity, damaging sightedness, and affecting quality of life. A corneal transplant is the current effective intervention. Due to the scarcity of donated cornea, such an unmet medical need requires a novel therapeutic modality. OBJECTIVES: Customizing patients' corneal endothelial progenitor cells with proliferative activity and lineage restriction properties shall offer sufficient therapeutic cells for corneal endothelial dystrophy. METHODS: The customized induced human corneal endothelial progenitor-like cell (iHCEPLC) was obtained through cell fate conversions starting from PBMC (peripheral blood mononuclear cell), hiPSC (human induced pluripotent stem cell), and hNCC (human neural crest cell), while it finally reached the iHCEPLC state via a series of induction. Several molecular diagnoses were applied to depict its progenitor state, including RNAseq, FlowCytometer, immunostainings, and rtPCR. Significantly, it can be induced to gain differentiation maturity through contact inhibition. In addition, a BAK-mediated rabbit model of corneal endothelial dystrophy was established in the present study to test the therapeutic effectiveness of the iHCEPLC. RESULTS: After inducing cell fate conversion, the specific HCEC markers were detected by rtPCR and immunostaining in iHCEPLC. Further, RNAseq was applied to distinguish its progenitor-like cell fate from primary human corneal endothelial cells (HECE). FlowCytometry profiled the heterogeneity subpopulation, consistently displaying a subtle difference from primary HCEC. A terminal differentiation can be induced in iHCEPLC, addressing its progenitor-like fate. iHCEPLC can restore the BAK-based rabbit model of corneal endothelial dystrophy. Immunohistochemistry verified that such acuity restoration of the BAK-treated cornea is due to the introduced iHCEPLC, and such therapeutic effectiveness is observed in the long term. CONCLUSION: Here, we demonstrated that customized iHCEPLC has long-term therapeutic efficacy. As a progenitor cell, our iHCEPLC has a restricted cell lineage nature and can proliferate in vitro, supporting sufficient therapeutic candidate cells. Due to the immune-privileged nature of the cornea, our iHCEPLC proves the principle of therapeutical feasibility in both autogenic and allogeneic modalities.

Assessment of iodine nutritional status and gestational thyroid function reference ranges during the first trimester of pregnancy in Taiwan.

BACKGROUND: Iodine nutrition is critical for fetal neurodevelopment in the first trimester of pregnancy, a period associated with dramatic changes in thyroid function. The aim of this study was to evaluate iodine nutritional status and thyroid function reference ranges in the first trimester in Taiwan. METHODS: Pregnant women aged 20 years and above in the first trimester were recruited in Taipei Veterans General Hospital, Taiwan from March 2019 to July 2022. Each participant provided a spot urine sample for measurement of urinary iodine concentration (UIC) and a blood sample for checkup of thyroid function and thyroid autoantibodies. A simple food frequency questionnaire was also completed. RESULTS: A total of 209 women with a mean age of 32.9 ± 4.4 years were enrolled. The median UIC was 160.9 µg/L (interquartile range [IQR]: 105.0-246.2 µg/L), indicating overall iodine sufficiency. The gestational thyroid function reference ranges were: thyroid stimulating hormone (TSH) (median: 0.93 [0.007-2.9] µIU/mL), free T4 (1.3 [0.93-2.2] ng/dL), free T3 (3.0 [2.3-5.0] ng/dL), total T4 (9.9 [6.4-16.9] ng/dL), and total T3 (135 [88-231] ng/dL). If the nonpregnant reference range of serum TSH was used, eight women (4.8%) would be misclassified as having subclinical hyperthyroidism, and two women (1.2%) with subclinical hypothyroidism would be missed. In multivariate analysis, nulliparous (adjusted odds ratio [OR] from model 1-3: 2.02, 2.05, 2.02; 95% CI, 1.08-3.77, 1.10-3.81, 1.11-3.66; p = 0.027, 0.023, 0.022, respectively) and multivitamin nonusers (adjusted OR from model 1-3: 1.86, 1.85, 1.78; 95% CI, 1.04-3.34, 1.03-3.32, 1.004-3.71; p = 0.038, 0.039, 0.049, respectively) had increased odds of having lower UIC levels <150 µg/L. CONCLUSION: The iodine nutritional status in the first trimester is adequate in Taiwan; however, certain subgroups such as nulliparous and multivitamin nonusers are still at risk for iodine deficiency. Gestational thyroid function reference ranges are needed for correct diagnosis of thyroid dysfunction in pregnancy.

[Mid-term clinical outcome of arthroscopic surgery on early knee osteoarthritis in middle-old aged patients].

OBJECTIVE: To compare the mid-term clinical effect of arthroscopic surgery versus conservative treatment on the middle aged early knee osteoarthritis (EKOA) patients, with the hope to provide clinical evidence for their individual therapy. METHODS: A total of 145 middle aged EKOA patients(182 knees) who received arthroscopic surgery or conservative treatment from January 2015 to December 2016 were retrospectively enrolled, including 35 males and 110 females, aged from 47 to 79 years old with an average of (57.6±6.9) years old, and the duration of disease ranged from 6 to 48 months with an average of(14.6±8.9) months. According to treatment method, patients were divided into arthroscopic surgery group (47 patients, 58 knees) and conservative treatment group(98 patients, 124 knees). Before treatment, patients presented with symptoms of knee joint, such as pain, swelling, locking, limited flexion and extension, and weakness, as well as abnormal findings in knee X-ray (without or suspicious joint space narrow, and a few of osteophyte formation) or in knee MRI (injury or degeneration of articular cartilage or meniscus, loose body in the joint cavity and synovial hyperemia edema, etc). Related data were collected, including duration of knee symptoms, presence of meniscus injury, loose body in the joint cavity or mechanical symptoms such as locking, and visual analogue scale (VAS) and Lysholm knee function score before treatment and at the latest follow-up. Statistical analysis was performed to compare the differences in VAS or Lyshilm score before or after treatment between the low groups and within each group. RESULTS: Patients in the two groups were followed up from 60 to 76 months. In the arthroscopic surgery group, the incision healing was good and no surgical complications occurred. There were no significant differences in age, gender, BMI and follow-up time between the two groups(P>0.05). Before treatment, compared with conservative group, duration of symptoms in the arthroscopic group was longer (P<0.001), comorbidity rates of meniscus injury (P<0.001), free body (P=0.001) and mechanical symptoms (P<0.001) were higher, VAS (P<0.001) and Lysholm score (P<0.001) were worse. At the final follow-up, VAS and Lysholm score in either the conservative group or the arthroscopic group were significantly better than before treatment (P<0.05), while no significant differences between the two groups were found. The VAS was (1.5±1.2) scores in the arthroscopic group and (1.6±1.0)scores in the conservative group(P=0.549), and the Lysholm score was (84.9±12.5) scores in the arthroscopic group and (84.2±9.9) scores in the conservative group (P=0.676). CONCLUSION: Both arthroscopic surgery and conservative treatment have satisfactory intermediate clinical effect middle- aged patients with EKOA, without statistically differences. However, most of the patients before surgery in the arthroscopic treatment group had mechanical locking symptoms caused by meniscus injury or loose body. Therefore, for the middle-aged EKOA patients with mechanical locking symptoms or without obtaining satisfactory outcome after conservative treatment, arthroscopic surgery may be considered.

[Diagnostic value of ultrasound in the discoid lateral meniscus of the knee].

OBJECTIVE: Based on the anatomical differences between discoid lateral meniscus(DLM) and normal lateral meniscus (NLM), this study aims to setting up the ultrasound examination parameters to distinguish DLM and NLM and explore the diagnostic value of these parameters on DLM. METHODS: According to the inclusion, exclusion and matching criteria, 66 DLM patients(DLM group) and 132 NLM patients with other knee joint diseases(NLM group), hospitalized from October 2019 to June 2020, were included in this study. There were 18 males and 48 females in the DLM group, ranging in age from 3 to 60 years old, with a mean of (36.9±12.1) years old;36 males and 96 females in the NLM group, ranging in age from 3 to 60 years old, with a mean of (40.0±12.2) years old. Philips high frequency(3.0 to 12.0 MHz) linear array ultrasound probe was used to examine the lateral meniscus of all patients in two groups. Nine parameters including the thickness, width and the included angle of the anterior angle, body and posterior angle respectively in the lateral meniscus were measured. The included angke is formed by the chords of the upper and lower curved surfaces of the meniscus at the free edge. The independent t test was used to analyze the difference of the measured parameters between the DLM group and the NLM group. The receiver operating characteristic(ROC) curve was used to evaluate the cut-off value of each parameter for diagnosing DLM and the corresponding sensitivity, specificity, positive predictive value, negative predictive value, and area under the ROC curve(AUC). RESULTS: The above 9 ultrasound measurement parameters between the DLM group and the NLM group had significant statistical differences(P<0.001). The ROC curve analysis method evaluated the cut-off value and diagnostic capabilities of these nine ultrasound parameters for DLM. The results showed that the cut-off value of the included angle of anterior part, body and posterior part were 25.85°, 24.85° and 29.15 °, respectively;and the sensitivity, specificity, negative predictive value, positive predictive value and AUC were significantly higher than other parameters, which were 88%, 91%, 79%, 95%, 0.94;89%, 94%, 82%, 97%, 0.96; 92%, 97%, 86%, 98%, 0.97, respectively. CONCLUSION: Ultrasound diagnosis of DLM is feasible and reliable. The diagnostic ability of the included angle of the anterior part, body and posterior part in the lateral meniscus measured by ultrasound to diagnose DLM are significantly better than other ultrasound measurement parameters, and the sensitivity and specificity of those parameters are close to MRI. Therefore, ultrasound can be used as a reliable method for preliminary diagnosis of DLM.

[Risk factors for the postoperative effect of symptomatic discoid lateral meniscus after arthroscopy].

OBJECTIVE: To explore the influencing factors of the postoperative effect of arthroscopic treatment of symptomatic discoid lateral meniscus (DLM). METHODS: From September 2008 to September 2015, patients with symptomatic DLM treated by arthroscopic surgery were retrospectively analyzed. The knee function was evaluated by Lysholm scoring system. According to the scoring results, it was divided into excellent (≥90 points), good (80 to 89 points), fair (70 to 79 points) and poor (<70 points). Sixteen research factors were collected, namely gender, operation age, body mass index, work intensity, symptom duration, history of knee injury, involved knee side, DLM classification, DLM injury type, DLM injury site, medial meniscus injury, knee cartilage injury site and degree, Kellgren-Lawrence (K-L) classification, operation method, and latest follow-up time. According to the data type, Kruskall-Wallis rank sum test or χ2 test was used for univariate analysis, and ordinal Logistic regression model was used for multivariate analysis. Univariate analysis showed that the statistically significant factors were included in the multivariate analysis model, and the independent risk factors influencing the curative effect of symptomatic DLM were obtained. RESULTS: According to the inclusion and exclusion criteria, 502 patients were included. Lysholm functional grade at the latest follow-up was higher than that before operation (P<0.001). Univariate analysis showed that there were significant differences in gender, body mass index, work intensity, history of knee injury, age of operation, duration of symptoms, medial meniscus injury, K-L grade, operation method, location and degree of knee cartilage injury among Lysholm functional grades(P<0.05). Ordinal logistic regression analysis showed that female was an independent risk factor for postoperative efficacy (P=0.002, OR=0.370);body mass index (P=0.007, OR=0.894), work intensity(P=0.004, OR=0.525), age of operation(P<0.001, OR=0.953), duration of symptoms(P<0.001, OR=0.989), degree of cartilage damage (P=0.016, OR=0.631) were negatively correlated with Lysholm knee function classification. CONCLUSION: Arthroscopy is a safe and effective method for the treatment of symptomatic DLM. The gender, body mass index, work intensity, age of operation, duration of symptoms and degree of knee cartilage injury are independent influencing factors for the postoperative efficacy of symptomatic DLM.

Multivariate ordered logistic regression analysis of the postoperative effect of symptomatic discoid lateral meniscus.

INTRODUCTION: The postoperative effect of arthroscopy in the treatment of symptomatic discoid lateral meniscus (DLM) varies greatly among individuals. Therefore, this study aims to investigate the factors affecting the postoperative outcomes of symptomatic DLM. MATERIALS AND METHODS: According to the inclusion and exclusion criteria, patients with symptomatic single-knee DLM who underwent arthroscopic surgery at our hospital from 9/2008 to 9/2015 were included. Retrospectively collected 16 factors probably affecting postoperative outcomes. The Ikeuchi grade system was used to evaluate the knee joint function. Univariate analysis was performed by Kruskal-Wallis rank-sum test or Mann-Whitney U test, and multivariate analysis by ordered logistic regression. P < 0.05 was considered statistically significant. RESULTS: A sum of 502 patients was included, including 353 females (70.3%) and 149 males (29.7%). Difference between preoperative and postoperative Ikeuchi grade was statistically significant (P < 0.001). Female was bad to obtain a good Ikeuchi grade (P = 0.009, OR 0.458). Outerbridge grade (P = 0.018, OR 0.638) was negatively correlated with Ikeuchi grade. BMI (P = 0.001, OR 0.875) and work intensity (P = 0.020, OR 0.611) were inversely correlated with Ikeuchi grade. Age of onset (P < 0.001, OR 0.956) and symptoms duration (P < 0.001, OR 0.988) were negatively correlated with Ikeuchi grade. Besides, compared to total meniscectomy, meniscoplasty with a repair was an unfavourable factor for Ikeuchi grade (P = 0.044, OR 0.245). CONCLUSIONS: With the increase of BMI, work intensity, age of onset, duration of symptoms, and the severity of cartilage lesion, the postoperative results become worse. Moreover, female and meniscoplasty with repair are risk factors for the postoperative outcomes.

A Reliable, Ultrasound-Based Method for the Diagnosis of Discoid Lateral Meniscus.

PURPOSE: To explore the feasibility and validity of ultrasound in the diagnosis of discoid lateral meniscus (DLM) by comparing quantitatively the morphologic difference between DLM and normal lateral meniscus. METHODS: This study was designed to develop and validate the ultrasound diagnostic criterion for DLM. In the development stage (July 2018 to June 2019), data from 180 subjects were used to derive the ultrasound diagnostic criterion, including 90 patients diagnosed as DLM by magnetic resonance imaging (DLM group) and 90 matched controls diagnosed as normal lateral meniscus diagnosed by magnetic resonance imaging (control group). Twelve distinct parameters of meniscus thickness, width, 0.5∗thickness/width, and angle were obtained through anterior, lateral, and posterior views with the probe oriented perpendicular to the lateral tibiofemoral joint line. In the validation stage (July 2019 to December 2019), data from 324 additional participants were used to validate the criterion derived from the development stage. Differences of the continuous variables and categorical variables between the 2 groups were analyzed by an independent t test and χ2 test, respectively. The diagnostic value of parameters was analyzed by the receiver operating characteristic curve. RESULTS: In the development stage, significant differences were found in the above 12 parameters between the 2 groups (P < .05 for all). Cut-off values of anterior meniscus angle, meniscus body angle, and posterior meniscus angle were 28.45°, 27.85°, and 29.15°, respectively. The area under the curve, sensitivity, and specificity of anterior meniscus angle (0.953, 95.6%, 91.1%), meniscus body angle (0.980, 95.6%, 95.6%), and posterior meniscus angle (0.942, 80.0%, 97.8%) were greater than other parameters. In the validation stage, the sensitivity, specificity, and accuracy of anterior meniscus angle, meniscus body angle, and posterior meniscus angle in diagnosing DLM were as high as 91.3%, 88.6%, and 89.2%; 94.2%, 93.3%, and 93.5%; and 76.8%, 95.7%, and 91.7%. CONCLUSIONS: DLM can be reliably diagnosed by ultrasound measurements of anterior meniscus angle, meniscal body angle, and posterior meniscus angle. LEVEL OF EVIDENCE: Level III, case-control study.

Factors influencing postoperative outcomes in patients with symptomatic discoid lateral meniscus.

BACKGROUND: Due to its abnormal morphology and ultrastructure, discoid lateral meniscus (DLM) is prone to tear and degeneration, leading to clinical symptoms. Arthroscopy is the main treatment for symptomatic DLM; however, postoperative outcomes vary widely due to the effects of diverse factors. This research aims to explore the factors influencing postoperative outcomes of symptomatic DLM. METHODS: Patients with DLM who underwent arthroscopic surgery at our hospital from 9/2008 to 9/2015 were enrolled according to the inclusion and exclusion criteria. Fourteen variables, including sex, body mass index (BMI) and other variables, were chosen as factors for study. Knee function was assessed using the International Knee Documentation Committee (IKDC) score. Univariate analyses (Mann-Whitney U test or Kruskall-Wallis rank sum test) and multivariate analyses (ordinal logistic regression) were used to identify the factors that influenced postoperative outcomes. P < 0.05 was considered statistically significant. RESULTS: A total of 502 patients, including 353 females (70.3%) and 149 males (29.7%), were enrolled. The median IKDC score postoperatively (87.4; range, 41.4 ~ 97.7; IQR, 14.6) was higher than that preoperatively (57.6; range, 26.9 ~ 64.9; IQR, 9.7) (P < 0.001). Male sex was predictive of a higher IKDC score (P = 0.023, OR = 1.702). Compared with BMI ≥25 kg/m2, < 18.5 kg/m2 was associated with better IKDC score (P = 0.026, OR = 3.016). Contrasting with age of onset ≥45 years, ≤14 years (P < 0.001, OR = 20.780) and 14 ~ 25 years (P < 0.001, OR = 8.516) were associated with better IKDC score. In comparison with symptoms duration> 24 months, IKDC scores for patients with symptoms duration ≤1 month (P = 0.001, OR = 3.511), 1 ~ 6 months (P < 0.001, OR = 3.463) and 6 ~ 24 months (P < 0.001, OR = 3.254) were significantly elevated. Compared to Outerbridge grade III ~ IV, no injury (P < 0.001, OR = 6.379) and grade I (P = 0.01, OR = 4.332) were associated with higher IKDC score. CONCLUSIONS: Arthroscopic treatment of symptomatic DLM is safe and effective, but its clinical efficacy is affected by many factors. Specifically, male sex, BMI < 18.5 kg/m2, age of onset < 25 years (especially < 14 years) and symptoms duration < 24 months are conducive to good postoperative outcomes. However, combined articular cartilage injury (Outbridge grade ≥ 2) reduces postoperative effect.

Impairment of Proteasome and Autophagy Underlying the Pathogenesis of Leukodystrophy.

Impairment of the ubiquitin-proteasome-system (UPS) and autophagy causing cytoplasmic aggregation of ubiquitin andp62 have been implicated in the pathogenesis of most neurodegenerative disorders, yet, they have not been fully elucidated in leukodystrophies. The relationship among impairment of UPS, autophagy, and globoid cell leukodystrophy (GLD), one of the most common demyelinating leukodystrophies, is clarified in this study. We examined the ubiquitin and autophagy markers in the brains of twitcher mice, a murine model of infantile GLD, and in human oligodendrocytes incubated with psychosine. Immunohistochemical examinations showed spatiotemporal accumulation of ubiquitin- and p62-aggregates mainly in the white matter of brain and spinal cord at disease progression. Western blot analysis demonstrated a significant accumulation of ubiquitin, p62, and LC3-II in insoluble fraction in parallel with progressive demyelination and neuroinflammation in twitcher brains. In vitro study validated a dose- and time-dependent cytotoxicity of psychosine upon autophagy and UPS machinery. Inhibition of autophagy and UPS exacerbated the accumulation of insoluble ubiquitin, p62, and LC3-II proteins mediated by psychosine cytotoxicity as well as increased cytoplasmic deposition of ubiquitin- and p62-aggregates, and accumulation of autophagosomes and autolysosomes. Further, the subsequent accumulation of reactive oxygen species and reduction of mitochondrial respiration led to cell death. Our studies validate the impairment of proteasome and autophagy underlying the pathogenesis of GLD. These findings provide a novel insight into pathogenesis of GLD and suggest a specific pathomechanism as an ideal target for therapeutic approaches.