[Research status of potential therapeutic targets for thyroid-associated ophthalmopathy].

Thyroid-associated ophthalmopathy (TAO) is an autoimmune eye disease that affects visual function and appearance, involving pathological remodeling processes of orbital tissue such as inflammatory reaction, oxidative stress, lipogenesis, and fibrosis. Current clinical first-line treatment options cannot be effective for all patients. This article summarizes the research on potential therapeutic targets of TAO at home and abroad in recent years, including receptor protein targets, immune cell targets, fat suppression targets, anti-fibrosis targets, transcription factor targets, and metabolic regulatory enzyme targets. Both non-natural compounds and natural compounds are introduced, with a view to providing clinical researchers with reference and ideas in the treatment of TAO and promoting the clinical application of new therapeutic drugs.

[Navigation-guided nasal endoscopy for removal of the cavernous hemangioma of the orbital apex through the sphenoid approach].

Objective: To explore the feasibility of navigation-guided nasal endoscopy for removal of the cavernous hemangioma of the orbital apex through the sphenoid approach. Methods: Retrospective case series study. From May 2012 to December 2019, 12 patients (12 eyes) with imaging findings of cavernous hemangioma in the orbital apex were collected at the Eye Hospital Affiliated to Nanchang University, including 3 males and 9 females aged 32 to 59 years. All patients underwent navigation-guided sinusoscopy through the sphenoid approach to remove the cavernous hemangioma of the orbital apex (video attached). Changes of visual function and complications after operation were analyzed. Results: In 3 patients without visual impairment, the postoperative visual function was still normal. Among the remaining 9 patients with preoperative visual impairment, visual function was fully recovered in 3 patients after operation, was improved in 2 patients, and had no change in 4 patients. There were no complications in 3 of the 12 patients, and 9 patients had transient, mildly limited intraocular rotation with diplopia after operation, which all returned to normal within 1 month. Conclusion: Navigation-guided sinus endoscopy through the sphenoid approach is effective and feasible in the removal of the cavernous hemangioma of the orbital apex. (Chin J Ophthalmol, 2021, 57: 837-843).

[Advances in the research and application of orbital blowout fracture repair material].

Orbital blowout fractures can easily lead to defects of the orbital wall. In order to restore the continuity of the bone wall and avoid a series of clinical symptoms caused by orbital contents herniation or incarceration, the site of the defect should be reconstructed. The effect of reconstruction depends on the choice of surgical plan and repair material. The typical materials for bone wall defect repair include bone sheet, high density porous polyethylene, titanium mesh, absorbable polymer, bioactive ceramics and tissue engineering bone. This paper reviews the research findings and application of material for repairing of orbital blowout fracture. (Chin J Ophthalmol, 2019, 55: 876-880).

[Application of three-dimensional printing combined with surgical navigation and endoscopy in orbital fracture reconstruction].

Objective: To evaluate the clinical effect of three-dimensional printing combined with surgical navigation and endoscopy for orbital fracture reconstruction. Methods: A case series study. Twenty-eight patients (28 eyes) with orbital fractures (20 males and 8 females, aged 10-61 years, with simple orbital fractures in 22 patients and composite orbital fractures in 6 patients) were treated with three-dimensional printing combined with surgical navigation and endoscopy for orbital fracture reconstruction at Affiliated Eye Hospital of Nanchang University from July 2016 to June 2018. With the help of three-dimensional printed models and guides, navigation positioning guidance and endoscopic visualization performance, the soft tissue incarcerated in the orbital fracture area was loosened, and the repair material was implanted (video attached). Postoperative follow-up was conducted at 1 week and 3 months. The follow-up observation included the best corrected visual acuity, diplopia, dyskinesia of the eyes, enophthalmos, and orbital volume. The data were analyzed by the paired t-test, Wilcoxon and the Mann-Whitney U rank sum test. Results: The best corrected visual acuity before and 1 week after surgery was 4.714±0.400 and 4.732±0.377, respectively, and the difference was not statistically significant (t=1.724, P=0.096). The enophthalmos before and 1 week after surgery was 2.2 (2.0-5.0) mm and 0.3 (0.0-2.3) mm, respectively, and the difference was significant (Z=-4.604, P<0.01). The orbital volume before and 1 week after surgery was 2 008.10 (6.84-11 200.00) mm(3) and 478.76 (5.01-7 286.00) mm(3), respectively, and the difference was statistically significant (Z=-3.735, P<0.01).The preoperative diplopia degree was 0, Ⅰ, Ⅱ, and Ⅲ in 14, 11, 3, and 0 eyes, while the degree of diplopia 3 months after surgery was 0, Ⅰ, Ⅱ, and Ⅲ in 22, 6, 0, and 0 eyes, respectively. The difference was statistically significant (Z=-2.359, P=0.018). The preoperative dyskinesia degree of the eyes was 0, Ⅰ, Ⅱ, and Ⅲ in 11, 11, 3, and 3 eyes, while the dyskinesia degree of the eyes 3 months after surgery was 0, Ⅰ, Ⅱ, and Ⅲ in 23, 5, 0, and 0 eyes, respectively. The difference was statistically significant (Z=-3.456, P=0.001). No implant infection, displacement, and other serious complications were observed during the follow-up of 3 to 12 months. Conclusions: Three-dimensional printing technology combined with nasal endoscopy and surgical navigation, which is applied in the reconstruction of orbital fracture, can significantly improve the symptoms of diplopia, ocular dyskinesia, and ocular depression. It is a feasible assistant method. (Chin J Ophthalmol, 2019, 55: 658-664).

[Analysis of the effect of different surgical methods on the cavernous hemangioma of the deep orbit].

Objective: To explore the effect of the surgical removal of orbital deep cavernous hemangioma following the concept of minimal surgical invasion. Methods: Retrospective case series study. Sixty-three cases with surgical removal of deep orbital cavernous hemangioma were collected in the Affiliated Eye Hospital of Nanchang University from May 2012 to August 2015. There were 29 males and 34 females. The age was from 17 to 69 years with mean age (45±11) years.The surgical methods the conjunctival approach orbital surgery, lateral orbital surgery, medial skin orbital surgery, lateral orbital conjunctiva conjunctival pathway and endoscopic nasal approach were chosen for different cases. The visual acuity, visual field and electrophysiological examination were analyzed before and after operation. Result: Sixty-three patients underwent complete surgically removal of the tumor. Among them, 32 cases were conducted by conjunctival approach, 24 cases had improved visual acuity, 5 cases had no change of visual acuity, 3 cases had decreased visual acuity. Seventeen cases performed with lateral open orbital surgery had improved visual acuity in 11 cases, no change in visual acuity in 3 cases and decreased visual acuity in 3 cases postoperatively. Six cases with approach of the medial orbital surgery had improved visual acuity in 4 cases, no change of visual acuity in 1 case and decreased visual acuity in 1 case postoperatively. Six cases with approach of outside open orbital surgery combined with medial conjunctival pathway had improved visual acuity in 4 cases, no change of visual acuity in 1 case and decreased visual acuity in 1 case postoperatively. Two cases performed with endoscopic nasal approach. One of them had normal visual function and no change after surgery. Another had impaired visual function pre-operatively and it came back to normal postoperatively. Conclusions: With the concept of minimal surgical invasion, the choice of appropriate surgical approach can save patients with the greatest degree of visual function and even improve the visual function of the patients with orbital cavernous hemangioma. (Chin J Ophthalmol, 2017, 53: 288-293).

Influences of biological risk at birth and temperament on development at toddler and preschool ages.

PURPOSES: This study aimed at investigating: (1) the effects of temperament and biological risk at birth on various developmental domains at toddler and preschool ages; (2) the interaction effects of the biological risk and temperament on development. Participants One hundred and ten full-term and 98 preterm children without significant physical or developmental disabilities and consisting of various biological risks were examined at toddler age (18-36 months) and preschool age (51-67 months). METHODS: The Neonatal Medical Index was used to classify the biological risk level. Parental reports on the Chinese Toddler Temperament Scale at toddler age were collected and the temperament (easy, intermediate and difficult) of each child was assigned according to local norm. The Comprehensive Developmental Inventory for Infants and Toddlers was used for assessing child development at toddler and preschool ages. Other family variables were also collected as possible confounders for child development. Two-way ancova was conducted to analyse the predictability of biological risk and temperament, by controlling the potential family variables on child development. RESULTS: At toddler age, higher biological risk had significant adverse effects on both the Perceptual-motor developmental quotients (DQs) (F(1,201)= 19.4, P < 0.001) and Social-adaptive DQs (F(2,200)= 22.5, P < 0.001), while easy temperament had positive effects only on the Social-adaptive DQs (F(2,200)= 7.7, P < 0.01). At preschool age, none of the two factors had effects on DQs of the Comprehensive Developmental Inventory for Infants and Toddlers. There were no significant interactions between biological risk and temperament on DQs at both ages. CONCLUSION: The biological risk and temperament affected child development at toddler age but not at preschool age. No interaction of biological risk and temperament effects on the child development at toddler age existed. The effects of biological risk and temperament on child development were temporary for the children with relatively low biological risk.

DNA methylation assay using droplet-based DNA melting curve analysis.

DNA methylation is an epigenetic regulation of gene expression, which has drawn great attention in biomedical research due to its association with various diseases. A robust, inexpensive platform to detect and quantify the methylation status in a specific genomic region is necessary. In this study, an on-chip analytical technique of cytosine methylation with droplets in a microchannel is proposed. Genomic DNA samples are encapsulated into a series of droplets and transported through a detection region, where a stable temperature gradient is created. As the temperature is elevated from 60 °C to 85 °C, the DNA samples denature and the associated fluorescence signals decay, with the relationship being acquired as the melting curve. The droplets serve as discrete reactors for conducting DNA melting curve analysis in the liquid phase, thereby eliminating the need for immobilization of reagents. Due to a high heating rate and greater enhanced thermal stability, this microchip allows larger melting temperature differences for the samples at different percentages of methylated DNA. It has an enhanced discrimination ability and lower volume consumption, compared to the commercial qPCR machine. This chip enables quantification of the methylation levels of the pluripotent stem cell factor Oct-4 in its distal enhancer (DE) region, with a designed probe after bisulfite treatment and asymmetric PCR.

Effect of a periodized exercise training and active recovery program on antitumor activity and development of dendritic cells.

AIM: Dendritic cells (DCs) are potent antigen-presenting cells that promote antitumor immunity in vivo when pulsed with tumor antigen. No studies have indicated that exercise training affects DC function. The purpose of this study was to investigate the effect of a 5-week periodized exercise training and active recovery program on the development of DCs, and to test their effect on the antitumor immunity of mononuclear cells (MNC) from blood and spleen against human leukemic U937 and murine lymphoma Yac-1 cells, respectively. METHODS: Male Fisher 344 rats were divided into 2 groups: exercise and non-exercise group. The training protocol consisted of running on a motor-driven treadmill 6 days a week for consecutive 5 weeks, during which the running time, treadmill speed, and incline gradient were increased weekly. Active recovery parameters were set at 30% of the intensity of the previous day. RESULTS: DC numbers increased significantly (P<0.05) in the exercise group compared to controls, but there were no significant changes in the expression of surface antigens CD80 and CD86. In exercise group MNC-conditioned medium (CM) prepared with 50 microg/mL phytohemagglutinin (PHA) significantly inhibited proliferation of U937 cells, and splenocyte-CM with PHA at 20 and 40 microg/mL significantly inhibited proliferation of YAC-1 cells greater than control group. CONCLUSIONS: The 5-week periodized exercise training with active recovery promotes the number of DCs and enhances the activity of DCs against tumor cells.

Serine 187 is a crucial residue for allosteric regulation of Corynebacterium glutamicum 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase.

Corynebacterium glutamicum 3-deoxy-D-arabino-heptulosonate-7-phosphate (DAHP) synthase is sensitive to feedback inhibition by tyrosine. One feedback-insensitive mutant was obtained by in vitro chemical mutagenesis and the mutation was identified as a C-->G mutation at nucleotide 560 causing a Ser(187) to Cys(187) substitution. Replacing Ser(187) with cysteine, tyrosine or phenylalanine by site-directed mutagenesis not only reduced the enzymatic activity but also relieved its feedback inhibition by tyrosine, while Ser(187)Ala exhibited a comparable activity to that of wild-type enzyme and sensitized to allosteric regulation. The His(6)-tagged enzymes were expressed in Escherichia coli and purified to homogeneity by immobilized nickel-ion affinity chromatography. Kinetic analysis showed that tyrosine is a competitive inhibitor of phosphoenol pyruvate, one of the precursors for DAHP biosynthesis.

Optimization of walking in children.

Previous work demonstrated that adults naturally adopt a walking frequency to optimize physiological cost, symmetry, and stability. Furthermore, the optimal frequency is predictable using the force-driven harmonic oscillator (FDHO) model. However, no studies have established the developmental processes of optimization in children. Thus, the purposes of this study were to examine the predictability of the preferred stride frequency (PSF) and optimization features of 3- to 12-yr-old children using the FDHO model. Forty-five children and nine adults were measured for anthropometric data to calculate the predicted frequency. They later walked at three frequencies (PSF, PSF +25%, and PSF -25%) at a constant speed on a treadmill. The results indicated that the FDHO model was accurate in predicting the preferred frequency of children (prediction error < 0.07 s). We identified three stages of learning in the development of optimization: an early manifestation of sensitivity to resonant frequency, the subsequent development of ability to modulate walking frequency, and the final establishment of an adult optimization form at age seven. Our findings suggest that walking development may be determined by the dynamic cooperation of physiological, neural, and musculoskeletal systems with respect to the environmental context.

Prognostic factors for walking attainment in very low-birthweight preterm infants.

The purpose of this study was to compare the age of walking attainment between very low-birthweight (VLBW) preterm infants and normal term infants, and to determine the variables that affect the walking attainment in VLBW infants. Ninety-six VLBW preterm infants and 82 normal term infants were prospectively followed to determine their age of walking attainment and to monitor gross motor development with sequential clinic visits at 6, 9, 12 and 18 months corrected age. Perinatal and sociodemographic data were collected through review of medical records. The VLBW infants were significantly older at attainment of walking (median 14 months) than the term infants (median 12 months) after correction for prematurity. By the age of 18 months, all term infants had attained walking ability; while 11% of VLBW infants were still unable to walk. Multivariate proportional hazards regression analysis revealed that low gestational age was significantly associated with late attainment of walking in VLBW infants. With the adjustment for gestational age, prolonged ventilation (or oxygen therapy) and severe retinopathy of prematurity were significant predictors of late walking attainment. Our findings indicate that VLBW preterm infants have an increased risk of delayed attainment of walking. Furthermore, the contribution of low gestational age to the delayed walking attainment in VLBW infants may occur via the plausible pathways of neonatal respiratory distress and severe retinopathy of prematurity.

Optimization of lipase-catalyzed biodiesel by response surface methodology.

Biodiesel prepared by catalyzed mild transesterification has become of much current interest for bioenergy. The ability of a commercial immobilized lipase (Novo Industries--Bagsvaerd, Denmark) from Rhizomucor miehei (Lipozyme IM-77) to catalyze the transesterification of soybean oil and methanol was investigated in this study. Response surface methodology and 5-level-5-factor central composite rotatable design were employed to evaluate the effects on reaction time, temperature, enzyme amount, molar ratio of methanol to soybean oil, and added water content on percentage weight conversion to soybean oil methyl ester by transesterification. Based on ridge max analysis, the optimum synthesis conditions giving 92.2% weight conversion were: reaction time 6.3 h, temperature 36.5 degrees C, enzyme amount 0.9 BAUN (Batch Acidolysis Units NOVO), substrate molar ratio 3.4:1, and added water 5.8%.

Locomotor strategies before independent walking: prospective study of 50 mentally retarded children.

To investigate the association between pre-walking locomotor strategies and psychomotor developments in children with mental retardation (MR), 50 children with non-specific MR were included in this study. There were 29 boys and 21 girls, 96% of whom had moderate to severe MR. They were followed from 4-53 months to 25-99 months of age, and their follow-up periods ranged from 10 to 48 months (mean 30 months). According to the pre-walking locomotor strategies, these children were categorized into three groups: the crawling group (n = 34) who used crawling or creeping as their main locomotion pattern before independent walking; the shuffling group (n = 9) who used shuffling prior to independent walking; and the direct-walking group (n = 7) who did not have any other locomotor strategies except rolling. In almost all motor developmental milestones, children in the direct-walking group developed earlier than those in the crawling and shuffling groups. Children in the crawling group had more advanced developments than those in the shuffling group. The difference in the mean ratio developmental quotients of the Bayley Mental Scale among the three groups was not significant. The present study showed that crawling may not be a necessary prerequisite for early ambulation or better cognitive function in MR children.

[Pharmacological study on Asparagus officinalis Linn].

Our experiment shows that Asparagus officinalis can increase the clearance rate of charcoal particles and the weight of immune organs in mice, and thus helps to facilitate their antifatigue, anoxia tolerance, analgesia and memory improvement, as well as decrease the contents of lipid peroxide (LPO) in plasma, liver and brain of the animal.

[Wind-damage effects on quality of heartwood of Lignum Santali Albi].

OBJECTIVE: To evaluate the wind-damage effects on quality of heartwood of Lignum Santali Albi. METHOD: GC-MS, TLC and pharmacodynamic test. RESULTS: The content of volatile oil from heartwood of Wind-damaged Lignum Santali Albi is 1.42%; the content of various components in the oil and the chromatography of different extracts are similar to those of reference drug and 25 years old trees. CONCLUSION: Wind-damage should accelerate the formation of heartwood of Lignum Santali Albi without influence on its quality.

G-CSF downregulates natural killer cell-mediated cytotoxicity in donors for hematopoietic SCT.

In G-CSF-mobilized hematopoietic SCT (HSCT), natural killer (NK) cells have a critical role in GVHD and GVL effects. However, regulation of NK cell response to G-CSF remains unclear. This study assayed G-CSF effects in both HSCT donors and NK-92MI cells. The donors who received G-CSF had significantly decreased NK cell cytotoxicity. Levels of phosphatidylinositol 3-kinase (PI3K) and phosphorylated (p)-Akt, but not mammalian target of rapamycin (mTOR), were downregulated in NK cells from G-CSF-injected donors. G-CSF also decreased cytotoxicity without affecting viability and NF-κB of NK-92MI cells. PI3K and p-ERK expression were also decreased in G-CSF-treated NK-92MI cells, and their inhibitors, wortmannin and PD98059, respectively, both enhanced the downregulation of cytotoxicity. These effects were accompanied by decreased expression of a cytotoxicity-related gene, triosephosphate isomerase (TPI). Wortmannin, but not PD98059, enhanced the downregulation of TPI in G-CSF-treated NK-92MI cells, indicating a correlation between PI3K and TPI. We conclude that G-CSF-impaired NK cell cytotoxicity may accompany PI3K/Akt signaling. The effect is transient and NK cells may recover after G-CSF clearance, suggesting that G-CSF-mobilized HSCT may benefit both acute GVHD prevention and late-phase GVL promotion in HSCT recipients.

Modulation of dendritic cells by endurance training.

A periodized endurance training is able to adapt physical and psychological needs and increase exercise performance. Dendritic cells (DCs) play a key role in regulation of the immune response. In the present study, we trained Sprague-Dawley (SD) rats for five weeks using a progressive endurance protocol with the aim of measuring the effect on myeloid DC differentiation and maturation. Rats were divided into a non-exercise group (NEG) and an exercise group (EG). Bone marrow cells were isolated from these rats after training and cultured in the presence of granulocyte/monocyte-colony stimulatory factor (GM-CSF) and interleukin (IL)-4, and the resultant immature DCs were triggered with lipopolysaccharide to mature. DCs were collected and the main characteristics of DCs were assessed. The recovery rate and the expression of major histocompatibility complex (MHC) class II molecules for DC collected from EG was markedly greater than NEG. The function of DCs from EG to trigger a mixed leukocyte reaction and IL-12 production was higher than NEG. There was no liver and renal toxicity observed in all rats. Changes in food/water consumption and body weight increase between the groups were normal for the conditions. This study demonstrated that periodized endurance training is able to modulate DC development and shift them towards a more mature state.

Macrophages derived from bone marrow modulate differentiation of myeloid dendritic cells.

Dendritic cells (DC) are specialized antigen-presenting cells. Bone marrow monocytes have been widely used to generate murine myeloid DC. We found that mouse macrophages derived from bone marrow CD11b+ monocytes influenced the differentiation of these precursors into DC. Modulation of differentiation was demonstrated by the down-regulation of CD11c, CD40, and CD86 expression and by IL-12 production. DC differentiated in the presence of conditioned medium from bone marrow-derived macrophage culture (MCM) had impaired ability to stimulate proliferation of, and IFN- gamma production by, allogeneic CD4+ T cells. This inhibition of DC differentiation was mainly mediated by secretory products from macrophages but not by cell-cell contact. MCM contained higher concentrations of macrophage-colony-stimulating factor (M-CSF), IL-10, and TGF- beta1, whereas IL-6 remained unchanged compared with conditioned medium from fresh monocytes. M-CSF may be the major mediator in MCM inhibiting DC differentiation. This study demonstrates an important influence of bone marrow-derived macrophages on DC precursors during DC differentiation.

Test-retest reliability of balance tests in children with cerebral palsy.

To investigate intrasession and intersession reliability of balance tests in children with or without disabilities, 50 children without disabilities (ND) and 36 children with cerebral palsy (CP) aged from 5 to 12 years were tested. Intrasession reliability of postural stability of the Smart Balance Master System and one-leg standing test were assessed in both groups and intersession reliability of the Smart Balance Master System and balance subtest of the Bruininks-Oseretsky Test of Motor Proficiency (BOTMP) were assessed in ND children. Intersession reliability of the postural stability test in ND children, obtained using the Smart Balance Master System, was of moderate to good reliability in centre target (CT), sway vision (SV), eyes open and sway surface (EOSS), and sway vision and sway surface (SVSS; ICC 0.72 to 0.84). In children with CP, intrasession reliability was high in CT (ICC 1). One-leg standing tests in both groups also had moderate to good intersession reliability (ICC 0.56 to 0.99). Agreement of failure score of lateral rhythmic shifting (LRS) at 1 second and 2 seconds pace was 85% and 93% respectively in ND children. Within the balance subtest of BOTMP, only two items had 100% agreement. Results suggest that postural stability tests in four conditions (CT, SV, EOSS, and SVSS), LRS, one-leg standing, and walking on a line are reliable and can be used to monitor balance control in ND children. Postural stability in CT condition and one-leg standing test are reliable in children with CP. Further study is needed to establish more reliable balance tests for children with CP.

Identification of essential cysteine residues in 3-deoxy-D-arabino-heptulosonate-7-phosphate synthase from Corynebacterium glutamicum.

To ascertain the functional role of cysteine residue in 3-deoxy-d-arabino-heptulosonate-7-phosphate (DAHP) synthase from Corynebacterium glutamicum, site-directed mutagenesis was performed to change each of the three residues to serine. Plasmids were constructed for high-level overproduction and one-step purification of histidine-tagged DAHP synthase. Analysis of the purified wild-type and mutant enzymes by SDS-polyacrylamide gel electrophoresis showed an apparent protein band with a molecular mass of approximately 45 kDa. Cys145Ser mutant retained about 16% of the enzyme activity, while DAHP synthase activity was abolished in Cys67Ser mutant. Kinetic analysis of Cys145Ser mutant with PEP as a substrate revealed a marked increase in Km with significant change in kcat, resulting in a 13.6-fold decrease in kcat/KmPEP. Cys334 was found to be nonessential for catalytic activity, although it is highly conserved in DAHP synthases. From these studies, Cys67 appears important for synthase activity, while Cys145 plays a crucial role in the catalytic efficiency through affecting the mode of substrate binding.