Use of MicroRNA Let-7 to Control the Replication Specificity of Oncolytic Adenovirus in Hepatocellular Carcinoma Cells

Highly selective therapy for hepatocellular carcinoma (HCC) remains an unmet medical need. In present study, we found that the tumor suppressor microRNA, let-7 was significantly downregulated in a proportion of primary HCC tissues (12 of 33, 36.4%) and HCC cell lines. In line with this finding, we have engineered a chimeric Ad5/11 fiber oncolytic adenovirus, SG7011let7T, by introducing eight copies of let-7 target sites (let7T) into the 3′ untranslated region of E1A, a key gene associated with adenoviral replication. The results showed that the E1A expression (both RNA and protein levels) of the SG7011let7T was tightly regulated according to the endogenous expression level of the let-7. As contrasted with the wild-type adenovirus and the control virus, the replication of SG7011let7T was distinctly inhibited in normal liver cells lines (i.e. L-02 and WRL-68) expressing high level of let-7 (>300 folds), whereas was almost not impaired in HCC cells (i.e. Hep3B and PLC/PRF/5) with low level of let-7. Consequently, the cytotoxicity of SG7011let7T to normal liver cells was successfully decreased while was almost not attenuated in HCC cells in vitro. The antitumor ability of SG7011let7T in vivo was maintained in mice with Hep3B xenograft tumor, whereas was greatly decreased against the SMMC-7721 xenograft tumor expressing a high level of let-7 similar with L-02 when compared to the wild-type adenovirus. These results suggested that SG7011let7T may be a promising anticancer agent or vector to mediate the expression of therapeutic gene, broadly applicable in the treatment for HCC and other cancers where the let-7 gene is downregulated

Proneural Genes in Neocortical Development

The timing of neocortical development is regulated by both intrinsic proneural genes Neurog2 and Ascl1, and extrinsic signals. However, how these intrinsic factors and extrinsic signals coordinatedly control neural cell fate decisions was not well understood at the start of my studies. Neurog2 and Ascl1 were known to be expressed by dorsal and ventral telencephalic progenitors, respectively. In the embryonic dorsal telencephalon, Neurog2 was known to promote the differentiation of glutamatergic projection neurons, while in the ventral telencephalon, Ascl1 was known to promote the formation of GABAergic interneurons and oligodendrocyte precursor cells (OPCs). However, little was understood about the regulatory controls that governed Neurog2 and Asc11’s proneural activities. During my PhD study, I first found that the proneural activity of Neurog2 declined at later stages of cortical neurogenesis, which was controlled by a simultaneous increase in GSK3 activity (Chapter 3). Furthermore I found that GSK3 inhibited Neurog2’s proneural activity through direct phosphorylation, which altered Neurog2-cofactor interactions. Next, I found that RAS/ERK signaling controlled a Neurog2-Ascl1 genetic switch, promoting Ascl1 expression while inhibiting Neurog2 expression (Chapter 4). Moreover, I found that RAS/ERK signaling modified Ascl1’s functions through direct phosphorylation, with higher levels of RAS/ERK causing Ascl1 to select OPC differentiation pathways, while lower levels allowed Ascl1 to promote a GABAergic neuronal fate. Notably, the interaction between RAS/ERK signaling and Ascl1 was found to be important in both normal neocortical development and in gliomagenesis. Finally, I observed that Neurog2 and Ascl1 proteins were co-expressed in a subset of early cortical progenitors, with the frequency of co-expression declining over developmental time (Chapter 5). Furthermore, I found that the Neurog2-Ascl1 co-expression is associated with enhanced neural stem cell properties and a decreased propensity to undergo neuronal differentiation. Accordingly, in Neurog2-/-;Ascl1-/- double mutant cortices, I found that neurogenesis and laminar marker expression was accelerated. I thus conclude that the Neurog2-Ascl1 co-expression ‘primes’ a subset of cortical progenitors to prevent lineage selection until later in development. Taken together, I made a great deal of progress in improving our understanding of how the proneural genes cooperate with extrinsic signals to regulate the timing of neocortical development

Hyperhomocysteinemia Associated with Multiple Organ Failure in Acute Pancreatitis Patients

Objective. This study aimed to evaluate the potential effect of hyperhomocysteinemia on multiple organ failure (MOF) in patients with acute pancreatitis (AP). Method. In this cohort study, a total of 1880 AP patients were enrolled and divided into the hyperhomocysteinemia group (study group) and the control group based on serum homocysteine (HCY) levels. Clinical data including demographics, clinical outcomes, and characteristics were collected for analysis. Risk factors of MOF in AP patients were determined by univariate and multivariate logistic regression analyses. Results. The hyperhomocysteinemia group showed higher multiple organ failure rates (31.83% vs 20.77%, P<0.001), compared with the control group. A positive correlation between homocysteine level and APACHE II score was obtained by Pearson correlation analysis (r = 0.420, P<0.001). Multivariate logistic regression analysis disclosed that the hyperhomocysteinemia was independently associated with MOF (hazard ratio, 1.103; 95% CI, 1.010–1.189; P=0.012). Conclusion. A high serum homocysteine level may be an independent risk factor of multiple organ failure in patients with acute pancreatitis

Herpetic Keratitis Preceded by COVID-19 Vaccination

The global Coronavirus Disease 2019 (COVID-19) pandemic has accelerated vaccine development at an unprecedented rate. A large population of people have received COVID-19 vaccines, while the vaccine safety data are limited. Here, we reported two cases of herpetic keratitis that occurred soon after receiving the inactivated COVID-19 vaccines. Case 1 was a 60-year-old woman who underwent penetrating keratoplasty (PKP) one year ago for corneal scarring caused by herpes simplex keratitis (HSK), and case 2 was a 51-year-old man with an unremarkable medical history. Both patients developed herpetic keratitis (HSK and varicella-zoster virus corneal endotheliitis, respectively) soon after receiving the inactivated COVID-19 vaccines (Sinovac). Herpetic keratitis was treated successfully with topical or plus oral antiviral ganciclovir. The short latency time in these two cases suggested that an inactivated COVID-19 vaccine may have a risk of triggering ocular herpes virus reactivation. Both clinicians and patients should be aware of this phenomenon. However, a causal relationship awaits confirmation

The inter-visit variability of retinal blood flow velocity measurements using retinal function imager (RFI)

Abstract Background To determine the inter-visit variability of retinal blood flow velocities (BFVs) using a retinal function imager (RFI) in healthy young subjects. Methods Twenty eyes of 20 healthy young subjects were enrolled. RFI imaging was performed to obtain the BFVs in retinal arterioles and venules in a field measuring 7.3 × 7.3 mm2 (setting: 35 degrees) centered on the fovea, and repeated measurements were obtained on two separate days. The inter-visit variability of BFVs was assessed by the concordance correlation coefficient (CCC) and coefficient of variance (CV). Results At the first visit, the mean BFV was 3.6 ± 0.8 mm/s and 3.0 ± 0.7 mm/s in arterioles and venules, respectively, which were not significantly different from those at the second visit (the BFV of arterioles was 3.5 ± 0.8 mm/s, and the BFV of venules was 3.0 ± 0.7 mm/s, P > 0.05, respectively). The CCC was 0.72 in the BFVs of arterioles and 0.67 in venules, and the CV was 10.8% in the BFVs of arterioles and 11.0% in venules. Conclusion The inter-visit variability using the retinal function imager (RFI) with a large field of view appeared to be good and comparable to previously reported intra-visit and inter-eye variability

The Prevalence of Demodex folliculorum and Demodex brevis in Cylindrical Dandruff Patients

Purpose. To compare the prevalence of and factors associated with Demodex brevis and Demodex folliculorum in patients with cylindrical dandruff (CD group) and healthy controls. Methods. Eyelashes were taken from 1680 patients with CD and 1700 healthy controls in China from March 2015 to May 2017. All patients underwent a complete eye examination, and Demodex spp. were counted. The prevalence was analyzed according to age, gender, and clinical features. Results. Mean patient age was 42.93 ± 16.52 (3–88) and 39.4 ± 13.6 (7–81) years old in the CD and healthy control groups, respectively. In the CD and healthy groups, the positive rate for Demodex folliculorum was 27.92% and 8.47%, respectively, while that for Demodex brevis was 31.67% and 6.65%, respectively. In the CD group, the prevalence of Demodex brevis was higher than that of Demodex folliculorum, no matter in the females (33.65% versus 29.01%) or the males (28.54% versus 23.88%) in the CD group. Moreover, the numbers of Demodex folliculorum and Demodex brevis were significantly and positively correlated with age, in both children and old patients (both P<0.001), as well as with the severity of eyelid congestion (all P<0.05). Conclusions. In a large sample population, the prevalence of Demodex brevis and Demodex folliculorum was higher in the CD group than in healthy volunteers. In addition, the severity of eyelid congestion might be exacerbated by the number of Demodex spp., which may therefore provide a good clinical reference and objective guide

Exposure to sunlight reduces the risk of myopia in rhesus monkeys.

Exposure to sunlight has recently been postulated as responsible for the effect that more time spent outdoors protects children from myopia, while early life exposure to natural light was reported to be possibly related to onset of myopia during childhood. In this study, we had two aims: to determine whether increasing natural light exposure has a protective effect on hyperopic defocus-induced myopia, and to observe whether early postnatal exposure to natural light causes increased risk of refractive error in adolescence. Eight rhesus monkeys (aged 20-30 days) were treated monocularly with hyperopic-defocus (-3.0D lens) and divided randomly into two groups: AL group (n=4), reared under Artificial (indoor) Lighting (08:00-20:00); and NL group (n=4), exposed to Natural (outdoor) Light for 3 hours per day (11:00-14:00), and to indoor lighting for the rest of the light phase. After being reared with lenses for ca. 190 days, all monkeys were returned to unrestricted vision until the age of 3 years. Another eight age-matched monkeys, reared with unrestricted vision under artificial lighting since birth, were employed as controls. The ocular refraction, corneal curvature and axial dimensions were measured before lens-wearing (at 23±3 days of age), monthly during the light phase, and at the age of puberty (at 1185+3 days of age). During the lens-wearing treatment, infant monkeys in the NL group were more hyperopic than those in the AL group (F=5.726, P=0.032). Furthermore, the two eyes of most NL monkeys remained isometropic, whereas 3 of 4 AL monkeys developed myopic anisometropia more than -2.0D. At adolescence, eyes of AL monkeys showed significant myopic anisometropia compared with eyes of NL monkeys (AL vs NL: -1.66±0.87D vs -0.22±0.44D; P=0.002) and controls (AL vs Control: -1.66±0.87D vs -0.05±0.85D; P<0.0001). All differences in refraction were associated with parallel changes in axial dimensions. Our results suggest that exposure to natural outdoor light might have an effect to reduced hyperopic defocus-induced myopia. Also, the data imply that early life exposure to sunlight may help to maintain normal development of emmetropization later in life, and thus lower the risk of myopic anisometropia in adolescent monkey

Rapid deterioration of Mooren’s ulcers after conjunctival flap: a review of 2 cases

Abstract Background Conjunctival flaps are a widely used treatment for numerous corneal ulcers that are caused by microorganismal infections. However, whether it can be performed on immune-mediated corneal ulcers is controversial. Case presentation We present two cases of Mooren’s ulcer that were treated using conjunctival flap in an attempt to prevent further corneal perforation at their local hospital. A rapid acceleration in ulcer progression was observed after a conjunctival flap was applied. Ultimately, the two patients underwent corneal transplantation, which required the postoperative use of topical immunosuppressants and resulted in a final cure. In the current report, we also discussed this incorrect surgical choice via a review of conventional interventions that are used to treat Mooren’s ulcer. Conclusions These two cases demonstrate that keratoplasty combined with topical immunosuppressants is effective in treating Mooren’s ulcer. Application of conjunctival flaps or autografting could promote progression of ulceration in Mooren’s ulcers

Preoperative descriptive statistics of subjects (Mean±SD).

<p>Preoperative descriptive statistics of subjects (Mean±SD).</p