Malignant glaucoma treated by low-dose laser cycloplasty: a 1-year multicenter prospective non-comparative study.

AIM: To report a one-year clinical outcomes of low-dose laser cycloplasty (LCP) among malignant glaucoma patients. METHODS: In this prospective, multicenter, non-comparative clinical study, participants with malignant glaucoma were recruited and underwent LCP at eight ophthalmic centers in China. Patients were followed up at 1wk, 1, 3, 6, and 12mo. Intraocular pressure (IOP), number of glaucoma medications, anterior chamber depth (ACD), and complications were recorded. Anatomical success was defined as the reformation of the anterior chamber based on slit-lamp biomicroscopy. Recurrence was defined by the presence of a shallow or flat anterior chamber after initial recovery from treatment. RESULTS: A total of 34 eyes received LCP. Mean IOP and medications decreased from 36.1±11.5 mm Hg with 3.3±1.5 glaucoma medications pre-treatment to 20.9±9.8 mm Hg (P<0.001) with 2.9±1.6 medications (P=0.046) at 1d, and 17.4±6.7 mm Hg (P<0.001) with 1.3±1.7 medications (P<0.001) at 12mo. The ACD increased from 1.1±0.8 mm at baseline to 1.7±1.0 mm and to 2.0±0.5 mm at 1d and 12mo, respectively. A total of 32 (94.1%) eyes achieved initial anatomical success. During follow-up, 2 (5.9%) eyes failed and 8 (23.5%) eyes relapsed, yielding a 12-month anatomical success rate of 64.3%. Complications including anterior synechia (8.82%), choroidal/ciliary detachment (5.88%) and hypopyon (2.94%) were observed within 1wk. CONCLUSION: LCP is simple, safe, and effective in reforming the anterior chamber in malignant glaucoma.

Varenicline solution nasal spray for dry eye disease in Chinese patients: a randomized phase 3 trial.

Background: Dry eye disease has a high prevalence and exerts a significant negative effect on quality of life. In China, there are currently no available nasal sprays to promote natural tear production in patients with dry eye disease. We therefore evaluated the efficacy and safety of OC-01 (varenicline solution) nasal spray versus vehicle in Chinese patients with dry eye disease. Methods: This was a randomized, multicenter, double-masked, vehicle-controlled, phase 3 clinical trial conducted at ophthalmology departments in 20 hospitals across China (NCT05378945). Eligible patients had a diagnosis of dry eye disease based on patient symptoms, Eye Dryness Score (EDS), Schirmer's Test (with topical anesthesia) Score (STS), and corneal fluorescein staining (CFS) score. Participants were randomly assigned 1:1 using an Interactive Web Response System (IWRS) to receive OC-01 0.6 mg/mL twice daily (BID) or vehicle nasal spray. Participants, investigators, and sponsor were all masked to treatment assignment. The primary endpoint was the percentage of subjects in the intention-to-treat population achieving ≥10 mm improvement in STS from baseline at week 4. Findings: In total, 340 patients were randomized from 21 July 2022 to 04 April 2023, 78.8% were female. Patients in the OC-01 group (n = 176) had significantly higher achievement of ≥10 mm improvement in STS (35.8% [n = 63] versus 17.7% [n = 29], stratified odds ratio: 2.67, 95% CI: 1.570-4.533, p = 0.0002) and a significantly greater increase from baseline STS (least-squares mean difference [SE]: 3.87 [0.794], p < 0.0001) at week 4 versus the vehicle group (n = 164). In addition, OC-01 led to a numerically greater reduction in mean EDS from baseline at week 4 compared to the vehicle group (LS mean [SE] difference: -1.3 [2.20]; 95% CI: -5.64 to 2.99, p = 0.5467). The most common adverse event was mild, transient sneezing (78% of OC-01 administrations). No serious adverse events related to nasal administration occurred. Interpretation: OC-01 (varenicline solution) nasal spray BID has clinically meaningful efficacy for reducing the signs (as measured by STS) and may improve the symptoms (as measured by EDS) of dry eye disease, with an excellent safety and tolerability profile, in the Chinese population. Funding: Jixing Pharmaceutical Co. Ltd.

Effect of virtual reality exercise on interventions for patients with Alzheimer's disease: A systematic review.

Virtual reality (VR) interventions are increasingly being used in rehabilitating and treating patients with neurological disorders. This study aimed to explore the effects of VR exercise interventions for patients with Alzheimer's disease (AD). A systematic review of the published literature on randomized controlled trials of VR technology applied to patients with AD was conducted using the preferred reporting entry for systematic reviews and Meta-analysis guidelines. Descriptive analyses were performed to assess the quality of the studies in terms of the characteristics of the included studies, samples, diagnoses, types of VR technologies, subjective and objective levels of immersion, and quality of studies. Eight studies were included, including a pooled sample of 362 patients with AD. A systematic review showed that most studies focused on patients with AD's cognitive and physical functions. The main finding was that VR interventions could help improve cognitive and physical balance in patients with AD. However, future studies should emphasize design and use well-accepted assessment tools to validate the effects of VR interventions further.

Does a home-based exercise program play any role in the treatment of knee osteoarthritis? A meta-analysis.

BACKGROUND: Knee-osteoarthritis is a very common joint disorder, affecting about 85% of the population worldwide. The effectiveness of home-based exercises is still debatable, with many studies indicating positive outcomes with few side effects, while others find them of limited utility. OBJECTIVES: To assess the role of home-based exercise (HBE) programs in the treatment of knee osteoarthritis. MATERIAL AND METHODS: Randomized controlled trials were included as per the predefined Population, Intervention, Comparison, Outcomes and Study (PICOS) criteria. Demographic summaries and event data for osteoarthritis therapy in the exercise and control groups were assessed, and comparative efficacy was evaluated using clustered graphs. The RevMan software was used to calculate the odds ratio (OR) and risk ratio of the included studies. The risk of bias was also evaluated and heterogeneity analysis was performed. RESULTS: Fifteen clinical trials performed from 2000 to 2022, with a total of 2922 osteoarthritis patients, were included in the study, according to the chosen inclusion criteria. We observed a reduction in Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) scores but a more marked improvement in clinical symptoms in the exercise group. The Knee Injury and Osteoarthritis Outcome Score (KOOS) increased only in the exercise group and not in the control group. We obtained a pooled OR of 0.59 (95% confidence interval (95% CI): 0.36-0.98), T2 value of 0.88, χ2 value of 185.41, degrees of freedom (df) value of 14, I2 value of 92%, and p-value <0.00001. The overall Z effect was 2.04 with a p-value of 0.04. The pooled risk ratio was 0.81 (95% CI: 0.66-0.99) with a T2 value of 0.14, χ2 value of 191.53, df value of 14, I2 value of 93%, and p-value <0.00001. CONCLUSION: The data from the studies included in this meta-analysis are in favor of the use of HBEs for the treatment of knee osteoarthritis.

Valsartan Regulates PI3K/AKT Pathways through lncRNA GASL1 to Improve Isoproterenol-Induced Heart Failure.

OBJECTIVE: This study is aimed at determining the expression and function of the GASL1 and PI3K/AKT pathways in isoproterenol- (ISO-) induced heart failure (HF). To determine the moderating effect of valsartan (VAL) on the progression of ISO-induced HF and to elucidate the related mechanism. MATERIALS AND METHODS: First, in in vivo experiment, we examined the effect of VAL on cardiac function in rats with ISO-induced HF. Similarly, quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were used to detect the effect of VAL on ISO-treated rat primary cardiomyocytes. Then, si-GASL1-transfected primary cardiomyocytes were constructed and Ad-si-GASL1 was injected through rat tail vein to achieve the effect of lowering GASL1 expression, so as to investigate the role of GASL1 in VAL's treatment of ISO-induced HF. RESULTS: In ISO-induced HF rat models, the GASL1 decreased while PI3K and p-AKT expressions were abnormally elevated and cardiac function deteriorated, and VAL was able to reverse these changes. In primary cardiomyocytes, ISO induces apoptosis of cardiomyocytes, and expression of GASL1 decreased while PI3K and p-AKT were abnormally elevated, which can be reversed by VAL. The transfection of primary cardiomyocytes with si-GASL1 confirmed that GASL1 affected the expression of PI3K, p-AKT, and the apoptosis of primary cardiomyocytes. Rat myocardium injected with Ad-si-GASL1 was found to aggravate the cardiac function improved by VAL. CONCLUSIONS: This study was the first to confirm that VAL improves ISO-induced HF by regulating the PI3K/AKT pathway through GASL1. And this study demonstrated a significant correlation between HF, VAL, GASL1, and the PI3K/AKT pathway.

Design and Methodology of a Multi-Centre Clinical Trial of Low Dose Laser Cycloplasty for the Treatment of Malignant Glaucoma in China.

PURPOSE: To summarize the design and methodology of a trial designed to evaluate the efficacy and safety of low dose laser cycloplasty (LCP) in treating malignant glaucoma. METHODS: Prospective, multicentre, non-controlled clinical trial. Subjects were recruited from eight ophthalmic centers in China. The target sample size was 34. Patients aged >18 years with a clinical diagnosis of malignant glaucoma inadequately controlled on medical therapy or malignant glaucoma recurrence after topical cycloplegics withdrawal were enrolled. All patients underwent LCP under retrobulbar anesthesia or sub-Tenon anesthesia. LCP is a treatment adopting few laser points (1100-2000 mW energy, 2000 milliseconds duration) that cauterizes and remodels the ciliary body over two clock hour-positions, which may relieve the ciliary ring block. Follow-up is planned for a period of 12 months. The primary outcome is the resolution of malignant glaucoma which is defined as central anterior chamber deepening after LCP. CONCLUSION: The Malignant Glaucoma Treatment trial (MGTT) will be the first prospective trial providing evidence of a treatment for malignant glaucoma. It intends to provide clinicians an optional, easy and convenient treatment for malignant glaucoma patients. Detailed morphological and biometric information collected during the study period will also help provide experience for the outcomes of malignant glaucoma. TRIAL REGISTRATION NUMBER: ChiCTR1800017960.

Measurement Properties of Canadian Agility and Movement Skill Assessment for Children Aged 9-12 Years Using Rasch Analysis.

The Canadian Agility and Movement Skill Assessment (CAMSA) was recently widely used to assess fundamental motor skills in children. Although the CAMSA is reported to be reliable and valid, its measurement properties are not clear. This study aimed to examine the measurement properties of the CAMSA in a sample of Chinese children using Rasch analysis. The study sample was from 1,094 children aged 9-12 years in Zunyi City, Guizhou Province. Descriptive data were analyzed using SPSS 24.0 software, and the dichotomous data were analyzed by Winsteps version 4.5.4 and Facets 3.67.1 software performing Rasch analysis. The present study investigated CAMSA measurement characteristics by Rasch analysis, including the reliability of the rating instrument, unidimensionality, item-fit statistics, and differential item functioning (DIF). Inter-rater reliability and retest reliability showed that the CAMSA had a good internal consistency. Rasch analysis indicated that the CAMSA was unidimensional, locally independent, and had a good item-fit-statistic. Additionally, the CAMSA displayed a good fit for the item separation index (12.50 > 2.0), as well as for item reliability (0.99 > 0.90). However, the item difficulty of the CAMSA did not fit well with personal ability, and a significant DIF was found across genders. In the Chinese children sample test, the CAMSA demonstrated appropriate goodness-of-fit validity and rater reliability. Thus, future research will explore item difficulty and person ability fit, as well as DIF across genders.

Evidence of Effect of Aerobic Exercise on Cognitive Intervention in Older Adults With Mild Cognitive Impairment.

This study aimed to evaluate the effectiveness of aerobic exercise as a cognitive intervention for older adults with mild cognitive impairment (MCI). The PubMed, EMBASE (Ovid), Cochrane Library, Web of Science, and Medline databases were searched from their inception until 30 April 2021. Randomized controlled trials (RCTs) examining the effects of aerobic exercise on global cognitive function in older adults with MCI were included. Ten eligible trials with acceptable methodological quality were identified. The meta-analysis results showed that aerobic exercise significantly improved the MMSE (N = 956, MD = 0.60, 95% CI: 0.28-0.92, p = 0.0003, I 2 = 31%, fixed effects model) and MoCA scores (N = 398, MD = 1.67, 95% CI. 1.18-2.15, p < 0.0001, I 2 = 37%, fixed-effects model) and overall cognitive performance in patients with MCI. The results of this study suggest that participation in regular aerobic exercise can improve cognitive function in older adults with MCI. These findings should be used with caution considering the limitations of the study.

Effect of Square Dance Exercise on Older Women With Mild Mental Disorders.

Many epidemiological studies have demonstrated the therapeutic benefits of exercise (EX) that can be used for adjunctive treatment in mental disorders. Despite several clinical experiments using exercise interventions, controlled studies are sparse in most disorder groups. Square dance is a popular aerobic exercise for older women in China. This study aimed to explore the effect of Chinese square dance exercise on mild mental disorders in older women. Participants included 109 older women with mild cognitive impairment from four large nursing homes. Participants were assigned either to the intervention group (n = 62) or the control group (n = 47), according to their residential nursing home. The intervention group underwent an 18-week square dance exercise, while the control group maintained their usual lifestyle. The outcomes were tested at baseline and weeks 9 and 18. The results showed that square dance exercise positively affected the results of all evaluations, especially on the participants' depressive symptoms and quality-of-life-related mental health. This study demonstrates that square dance exercise is a safe and effective approach for older women with mild cognitive impairment that benefits their long-term health.

The Effect of Chinese Square Dance Exercise on Cognitive Function in Older Women With Mild Cognitive Impairment: The Mediating Effect of Mood Status and Quality of Life.

The present study aimed to assess the effects of square dance exercise on the cognitive function and quality of life in older women with mild cognitive impairment and to investigate the mediating role of a depressed mood and reduced quality of life in the relationship between square dance exercise and cognition. The study design was a single-blind whole-group controlled trial. A total of 136 eligible participants were divided according to their nursing home into either an experimental or control group. The nursing home grouping was determined by the drawing of lots. The Montreal Cognitive Assessment (MoCA), Quality of Life (SF-12) and Geriatric Depression Scale (GDS-15) were used to assess participants at baseline, week 9, and week 18, respectively. Generalized estimating equations (GEE) were used to compare the results at baseline with mid-test and post-test changes in cognitive function and quality of life. Maximum likelihood estimation (ML) and robust standard errors were used to perform the mediation model. The study results indicated that the experimental group (compared to the control group) had a significant improvement in cognitive function, quality of life, and mood state at baseline in the mid-test and post-test results. The results of this 18-week experiment showed that the exercise-cognition relationship was significantly mediated by a reduction in depressive symptoms (indirect effect: ß = -0.375; 95% CI = -0.864 to -0.069) and an improvement in quality of life (indirect effect: ß = -0.678; 95% CI = -1.222 to -0.290). This study revealed the effects of moderate-intensity square dance exercise on cognitive function and quality of life in older Chinese women with mild cognitive impairment and explored the potential mediating mechanisms. These findings can be used to inform the development of public health policies to promote brain health in older adults with mild cognitive impairment.

A Multicenter Study of the Prevalence of Dry Eye Disease in Chinese Refractive Surgery Candidates.

OBJECTIVE: The aim of this study was to determine the prevalence of preoperative dry eye disease and evaluate tear film function in refractive surgery candidates in China. METHODS: In this prospective noninterventional cross-sectional study, refractive surgical candidates from 13 preselected eye hospitals in China were recruited from July 2015 to February 2016. Patient histories, subjective symptoms, tear film breakup time (TBUT), ocular surface fluorescein staining, and Schirmer I tests (SIT), were assessed to conduct subgroup analysis. RESULTS: A total of 1,849 patients were recruited, 41.4% were diagnosed with dry eye disease (766/1,849) and 44.9% (830/1,849) of subjects had a positive history of contact lens (CL) wear. The overall mean TBUT and SIT values were 7.3 ± 3.7 s and 15.2 ± 8.8 mm, respectively. The total prevalence of ocular surface fluorescein staining was 23.46% (422/1,849); 44.62% of patients had TBUT <5 s and 23.20% of patients had SIT <5 mm. CL wearers were observed to have a higher prevalence of dry eye than non-CL wearers (54.1 vs. 35.2%, OR = 2.17, 95% CI: 1.77-2.65). CONCLUSIONS: In this study, the most common abnormal finding in dry eye disease was tear film instability. A high proportion of refractive surgery candidates have preexisting dry eye disease and a history of CL wear prior to surgery. Careful attention should be given to the evaluation of preoperative dry eye in refractive surgery candidates.

Hypoxia training improves hepatic steatosis partly by downregulation of CB1 receptor in obese mice.

Hypoxia training (HT) can reduce body weight and improve fatty liver. However, the mechanism is not clear. A previous study indicated that HT-induced weight loss might be associated with the endocannabinoid system (ECS), which has also been reported recently to be involved in the persistent lipid mediators after weight loss. The present study investigated the effects of HT, a new prospective weight-loss method, on nutritionally obese mice and demonstrated that HT significantly reduced body weight, fat mass, transcriptional expression of liver endocannabinoid receptor 1 (CB1), biosynthetic enzyme diacylglycerol lipase α (DAGLα) and improved the transcriptional expression of degrading enzyme monoacylglycerol lipase (MAGL). Liver endocannabinoids 2-arachidonoylglycerol (2-AG) but not anandamide (AEA) was evidently decreased in response to HT. Simultaneously, HT significantly reduced liver index, serum alanine aminotransferase (ALT) and liver fat contents. Western blot showed decreased expression of liver CB1, sterol regulatory element-binding protein-1 (SREBP-1), peroxisome proliferator-activated receptor γ (PPARγ) and increased expression of adipose triglyceride lipase (ATGL) and carnitine palmitoyltransferase-1 (CPT-1) levels after HT. However, intraperitoneal injection of CB1 receptor agonist WIN55212-2 offset the benefits by which HT reduced hepatic fat synthesis, with significant increased protein expression of SREBP-1 and PPARγ. Taken together, these findings reported the alleviation of obesity and hepatic steatosis through HT and provided a putative molecular mechanism by inhibiting the CB1-mediated fat synthesis.

Enriched environment improves working memory impairment of mice with traumatic brain injury by enhancing histone acetylation in the prefrontal cortex.

Working memory impairment is a common cognitive dysfunction after traumatic brain injury (TBI), which severely affects the quality of life of patients. Acetylcholine is a neurotransmitter which is closely related to cognitive functions. In addition, epigenetic modifications are also related to cognitive functions. A neurorehabilitation strategy, enriched environment (EE) intervention, has been widely used to improve cognitive impairment. However, studies of the mechanism of EE on cholinergic system and epigenetic modifications in mouse with TBI have not been reported yet. In this paper, a mouse model with traumatic frontal lobe injury was established, and the mechanism on EE for the mice with TBI was explored. It was found that EE could improve Y-maze performance of mice with TBI, the function of cholinergic system, and the imbalance of acetylation homeostasis in the prefrontal cortex of contralateral side of TBI. In addition, EE also could increase the level of CREB binding protein and histones H3 acetylation at ChAT gene promoter region in the prefrontal cortex of contralateral side of TBI. These indicate that EE has an important effect on the improvement of working memory impairment and the underlying mechanism may involve in histones H3 acetylation at ChAT gene promoter regions in the prefrontal cortex.

The use of concentrated monosodium glutamate wastewater as a conditioning agent for adjusting acidity and minimizing ammonia volatilization in livestock manure composting.

In this study, concentrated monosodium glutamate waste (CMGW) was proposed as a conditioning agent to adjust acidity and decrease ammonia (NH3) volatilization in thermophilic aerobic composting based on two incubation experiments. The results showed that with the addition of CMGW, NH3 volatilization of compost mixture under high temperature phase decreased significantly and pH met the current national standard within 5.5-8.5. When CMGW dosage increased to 2% (v/w), the decrease in NH3 volatilization was as high as 78.9%. This effect was enhanced by repeated application of CMGW. Furthermore, although the electrical conductivity increased with the application of CMGW, both the germination index and the microbial respiration of compost mixture implied that CMGW had no negative effects on the maturity of compost, instead, a comprehensive maturity might be accelerated. It was concluded that CMGW was an optional conditioning agent for thermophilic aerobic composting of livestock manure in regards to adjusting acidity and preventing nitrogen loss from NH3 volatilization.

[Clinical research of electroacupuncture combined with transperineal injection of BTX-A for neurogenic bladder after spinal cord injury].

OBJECTIVE: To evaluate the clinical efficacy of electroacupuncture (EA) combined with transperineal injection of botulinum toxin-A (BTX-A) on neurogenic bladder caused by spinal cord injury. METHODS: One day af ter surgery, 35 cases of spinal cord injury accompanied with neurogenic bladder were randomly divided into a BTX-A plus EA group (20 cases, group A) and a BTX-A group (15 cases, group B). The two groups were both treated with regular rehabilitation training of bladder function and injection of 200 IU (4 ml) BTX-A through perineum external urethral sphincter; the group A was additionally treated with EA at Zhongji (CV 3), Guanyuan (CV 4), Shenshu (3BL 23), Huiyang (BL 35) and Baliao (Shangliao (BL 31), Ciliao (BL 32), Zhongliao (BL 33), Xialiao (BL 34)), once a day, 40 min per treatment. The treatment was given 6 times per week for 4 weeks. The urination status in two groups before and after treatment was observed, and urodynamics examination and urethral pressure test were also made. RESULTS: After 4-week treatment, mean times of urinary incontinence, mean urethral catheter output, pressure of bladder and volume of urinary incontinence were all improved in two groups (all P<0.05), which were more significant in the group A (all P<0.05). The residual urine, maximum bladder capacity, maximum urethral closure pressure and maximum urine flow rate were all improved in two groups after treatment (all P<0.01); the improvement of residual urine, maximum bladder capacity, maximum urethral closure pressure in the group A was more significant than that in the group B (all P<0.05). CONCLUSION: Electroacupuncture com bined with transperineal injection of BTX-A could effectively improve the urination dysfunction in patients with neurogenic bladder after spinal cord injury.

In vitro antioxidant activity and in vivo antifatigue effect of layered double hydroxide nanoparticles as delivery vehicles for folic acid.

Folic acid antioxidants were successfully intercalated into layered double hydroxides (LDH) nanoparticles according to a previous method with minor modification. The resultant folic acid-LDH constructs were then characterized by X-ray powder diffraction and transmission electron microscopy. The in vitro antioxidant activities, cytotoxicity effect, and in vivo antifatigue were examined by a series of assays. The results showed that folic acid-LDH antioxidant system can scavenge 1,1-diphenyl-2-picrylhydrazyl and hydroxyl free radicals and chelate pro-oxidative Cu(2+). The in vitro cytotoxicity assays indicated that folic acid-LDH antioxidant system had no significant cytotoxic effect or obvious toxicity to normal cells. It also prolonged the forced swimming time of the mice by 32% and 51% compared to folic acid and control groups, respectively. It had an obvious effect on decreasing the blood urea nitrogen and blood lactic acid, while increasing muscle and hepatic glycogen levels. Therefore, folic acid-LDH might be used as a novel antioxidant and antifatigue nutritional supplement.

Effect of apigenin on gap junctional intercellular communication in human Tenon's capsule fibroblasts.

PURPOSE: To investigate the effect of apigenin on gap junctional intercellular communication (GJIC) in human Tenon's capsule fibroblasts (HTFs) and its underlying mechanism. METHODS: After a 48 h treatment of cultured HTFs with apigenin (80 micromol/L), the GJIC was detected by a scrape-loading/dye transfer technique with Lucifer yellow dye and rhodamine (Rh) dextran. The coupling index represents a quantification of GJIC where a high coupling index is associated with a greater number of cells demonstrating cell-cell communication through gap junction channels. The changes in connexin 43 (Cx43) distribution and the expression of Cx43 at the protein and mRNA levels were statistically compared between the two groups by means of immunocytochemistry, western blotting, and real-time polymerase chain reaction (PCR). RESULTS: The functioning of GJIC in the HTFs was significantly enhanced after 48 hours by apigenin treatment when compared with the control cells. In the apigenin group, the intercellular dye transfer grade was above 9, while this value was only grade 3-4 in the control group. The coupling index was significantly increased up to 9.205+/-0.3621 in the apigenin group, compared with 5.1775+/-0.3177 in the control group (F=279.581, P=0.000). The expression of Cx43 at the protein and mRNA levels was significantly up-regulated in the apigenin group compared with the control group. CONCLUSION: Apigenin can significantly enhance the function of GJIC in HTFs by up-regulating the expression of Cx43 at both the protein and mRNA levels, suggesting that the enhancement of GJIC in HTFs by apigenin probably acts as an important mechanism underlying the inhibitory effect of apigenin on HTF proliferation.

[Anti-proliferative effect of herpes simplex virus thymidine kinase gene system on human Tenon capsule fibroblasts in vitro].

OBJECTIVE: To observe anti-proliferative effect of herpes simplex virus thymidine kinase gene system (HSV-tk) on human Tenon capsule fibroblasts (HTFs) and it's mechanism. METHODS: Retroviral vector was constructed containing HSV-tk gene and transfer to packaging cell line PT67. The positive clones was selected with G418 and the supernatant was collected which contains virus particles. The titer of the virus was also calculated. The virus particles were verified by transmission electron microscope (TEM). PCR was carried out to detect the integrity of tk gene into viral DNA genome. HTFs cells were infected with the virus and the positive clones were selected for propagation culture. RT-PCR and Western blot were used to detect tk gene expression. The anti-proliferative effect of tk gene on HTFs cells was determined by MTT. Apoptosis of cells was detected by Hoechst 33258 DNA staining and flow cytometry. The morphologic changes of the cell were observed by TEM. RESULTS: The retroviral vector pL (tk) SN was constructed successfully which was verified by enzyme cutting. The shape of virus particles derived from the package cells appeared to be oval or spherical under TEM with the diameter around 100 nm. The tk gene was detected in the viral DNA genome by PCR. The virus titer was 4 x 10(7) cfu/ml. The expression of tk gene was detected by RT-PCR and Western blot both in mRNA and protein level. The anti-proliferative effect of tk gene on HTFs cells was dose-dependent, all cells were died 5 days after 5 x 10(-3) g/L GCV being added, IC50 = 6 x 10(-4) g/L. Both necrosis and apoptosis were observed in this study and the apoptosis rate was increased with increasing dose of GCV. CONCLUSIONS: The proliferation of HTFs cells in vitro could be effectively inhibited by the killing effect of HSV-tk-GCV system through the pathway of necrosis and apoptosis.