ObjectiveTo analyze the microbial culture positivity rate， pathogen spectrum， prevalence of multidrug-resistant organisms （MDROs）， and differences among donor procurement sources in corneas from the Eye Bank of Guangdong Province， and to provide evidence for eye bank quality control and the prevention of post-keratoplasty infections. MethodsThis retrospective study included 7 442 donor corneas procured by the Eye Bank of Guangdong Province and used for keratoplasty at Zhongshan Ophthalmic Center， Sun Yat-sen University， between 2018 and 2025. During surgery， donor corneoscleral rim tissue was excised for bacterial and fungal culture. Culture-positive specimens underwent microbial identification and antimicrobial susceptibility testing （AST）. Culture-positive corneas were compared across study years， pathogen categories， and donor procurement sources. ResultsOf the 7 442 donor corneas， 251 were culture-positive， corresponding to an overall culture positivity rate of 3.37%， with annual rates ranging from 1.50% to 5.31%. Among culture-positive corneas， 178 （70.92%） yielded bacteria alone， 72 （28.68%） yielded fungi alone， and 1 （0.40%） yielded both bacteria and fungi. Candida spp. and Aspergillus spp. were the predominant fungal isolates. Gram-negative bacteria were more frequently isolated than Gram-positive bacteria. The most common bacterial isolates were Acinetobacter baumannii， Staphylococcus epidermidis， Stenotrophomonas maltophilia， Enterococcus faecium， and Pseudomonas fluorescens. MDROs accounted for 75.47% of bacterial isolates， predominantly Acinetobacter baumannii， Staphylococcus epidermidis， and Enterococcus faecium， and demonstrated an increasing trend over the study period. Donor corneas procured through the Hospital Cornea Retrieval Program （HCRP） had a higher culture positivity rate than those obtained through Organ Procurement Organization （OPO） （P＜0.000 1）. Among culture-positive donor corneas， the proportion of fungal culture-positive corneas was significantly higher in the HCRP group than in the OPO group （P=0.001 5）， whereas the proportion of MDROs among bacterial culture-positive corneas was significantly higher in the OPO group than that in the HCRP group （P=0.005 2）. ConclusionsThe overall culture positivity rate of donor corneas from the Eye Bank of Guangdong Province is relatively low， indicating that the risk of microbial contamination is controlled. However， pathogen profiles and antimicrobial resistance patterns varies by procurement source. Ongoing microbiological and antimicrobial resistance surveillance， together with optimized donor risk assessment and timely reporting of culture results， may further improve eye bank quality management and help inform strategies for preventing post-keratoplasty infections.

ObjectiveTo analyze the microbial culture positivity rate， pathogen spectrum， prevalence of multidrug-resistant organisms （MDROs）， and differences among donor procurement sources in corneas from the Eye Bank of Guangdong Province， and to provide evidence for eye bank quality control and the prevention of post-keratoplasty infections. MethodsThis retrospective study included 7 442 donor corneas procured by the Eye Bank of Guangdong Province and used for keratoplasty at Zhongshan Ophthalmic Center， Sun Yat-sen University， between 2018 and 2025. During surgery， donor corneoscleral rim tissue was excised for bacterial and fungal culture. Culture-positive specimens underwent microbial identification and antimicrobial susceptibility testing （AST）. Culture-positive corneas were compared across study years， pathogen categories， and donor procurement sources. ResultsOf the 7 442 donor corneas， 251 were culture-positive， corresponding to an overall culture positivity rate of 3.37%， with annual rates ranging from 1.50% to 5.31%. Among culture-positive corneas， 178 （70.92%） yielded bacteria alone， 72 （28.68%） yielded fungi alone， and 1 （0.40%） yielded both bacteria and fungi. Candida spp. and Aspergillus spp. were the predominant fungal isolates. Gram-negative bacteria were more frequently isolated than Gram-positive bacteria. The most common bacterial isolates were Acinetobacter baumannii， Staphylococcus epidermidis， Stenotrophomonas maltophilia， Enterococcus faecium， and Pseudomonas fluorescens. MDROs accounted for 75.47% of bacterial isolates， predominantly Acinetobacter baumannii， Staphylococcus epidermidis， and Enterococcus faecium， and demonstrated an increasing trend over the study period. Donor corneas procured through the Hospital Cornea Retrieval Program （HCRP） had a higher culture positivity rate than those obtained through Organ Procurement Organization （OPO） （P＜0.000 1）. Among culture-positive donor corneas， the proportion of fungal culture-positive corneas was significantly higher in the HCRP group than in the OPO group （P=0.001 5）， whereas the proportion of MDROs among bacterial culture-positive corneas was significantly higher in the OPO group than that in the HCRP group （P=0.005 2）. ConclusionsThe overall culture positivity rate of donor corneas from the Eye Bank of Guangdong Province is relatively low， indicating that the risk of microbial contamination is controlled. However， pathogen profiles and antimicrobial resistance patterns varies by procurement source. Ongoing microbiological and antimicrobial resistance surveillance， together with optimized donor risk assessment and timely reporting of culture results， may further improve eye bank quality management and help inform strategies for preventing post-keratoplasty infections.

Objective:To study the difference in the extraction efficiency of the novel coronavirus (2019-nCoV) nucleic acid by using magnetic beads method, centrifugal column method and one-step method.Methods:On March 5, 2021, 10 throat swabs were collected from the staff working in the nucleic acid sampling room in Department of Clinical Laboratory, Affiliated Taikang Xianlin Drum Tower Hospital, Medical School of Nanjing University. The positive quality control samples were mixed into the swabs and used as mock positive samples. The RNA was extracted from simulated positive samples and their diluted samples by using magnetic beads method, centrifugation column method and one-step method. The purity ( A260/ A280 ratio) and concentration of the nucleic acid obtained were measured by micro-uv photometry, and fluorescence quantitative PCR was performed to compare the CT value and extraction efficiency. The three methods were used to extract the simulated weak positive specimens and to compare the difference of CT values after amplification. The measurement data that followed normal distribution were expressed by xˉ±s, the t test was used for comparing in the same group, and single factor analysis of variance was used for comparing among multiple groups. A P value smaller than 0.05 indicated a significant difference. Results:2019-nCoV nucleic acid extracted by magnetic bead method, centrifugal column method and one-step method could amplify positive results. There was no significant difference between the CT value of RNA amplification extracted by magnetic bead method and one-step method ( t=? 0.995 , P=0.376). The CT values of orf1ab gene amplified by centrifugal column method, magnetic bead method and one-step method were 29.28±0.06, 30.82±0.14 and 29.79±0.01 respectively ( F=11.196 , P=0.041). The CT values of E gene were 28.52±0.40, 27.33±0.78 and 27.38±0.13 respectively ( F=3.407, P=0.169). The CT values of N gene were 28.61±1.02, 27.24±0.20 and 27.25±0.47, respectively ( F=2.880 , P=0.020). The CT values of human genes extracted by centrifugal column method, magnetic bead method and one-step method were 19.68±0.36, 20.14±0.06 and 20.58±0.49 respectively, which was statistically significant ( F=4.904, P=0.048). The CT value of amplified human gene was affected by the dilution of human samples twice. The CT value of undiluted samples was smaller than that of diluted samples twice, with a difference of 2.95±0.22, which was statistically significant ( t=?3.025, P=0.039). The extraction time of one-step method, magnetic bead method and centrifugal column method were (15.00±1.50), (20.00±1.50) and (40.00±5.5) min respectively, and the difference was statistically significant ( F=688 , P=0.027). Conclusions:Magnetic bead method, centrifugal column method and one-step method can be used to extract 2019-nCoV nucleic acid, for the centrifugal column method has a higher extraction efficiency than the magnetic bead method and the one-step method. The one-step method is the fastest, followed by the magnetic bead method and the centrifugal column method. A large number of clinical samples can be processed using the magnetic bead method and one-step method. One-step rapid nucleic acid test can also be performed on samples from emergency and fever clinics. It is not recommended to dilute specimens for testing. In order to improve the detection rate, extracting RNA from highly suspected samples with negative initial nucleic acid test by centrifugal column method is suggested.

Objective:To explore the clinical efficacy of single-center infant kidney donor adult dual kidney transplantation to explore the difference of different operation methods and the operation options of different donor kidney conditions so as to improve the success rate of children kidney donor adult dual kidney transplantation and reduce complications.Methods:A total of 42 cases of infant and adult dual kidney transplantations at Department of Kidney Transplantation in The Second Xiangya Hospital of Central South University from December 2012 to May 2019 were divided into two groups according to whether or not donor kidney fulfilled the criteria of three " 5" . According to different surgical approaches, they were divided into three groups of A (classical En-Bloc operation), B (separated dual kidney transplantation) and C (modified operation). The clinical data and prognoses were analyzed.Results:The median follow-up period was 55(11-92) months. The estimated glomerular filtration rate was 123.4(92.2-156.6) ml/min for operation A, 97.2(81.3-116.6) ml/min for operation B and 133.9(133.9-133.9) ml/min for operation C. In donor group not fulfilling the " 5" principle, no thrombotic event occurred for operation A/C and 3 cases of transplantation for operation B. There were single renal embolism ( n=2) and dual renal embolism ( n=1)(3/5, 60%)( P<0.05). Urinary protein was positive in the last follow-up: operation A (1/2, 50%) and operation B (3/5, 60%) ( P<0.05). The estimated glomerular filtration rate at the last follow-up was 82.4(80.9-83.9) ml/min for operation A, 71.8(46.1-114.2) ml/min for operation B and 122(83.3-142.4) ml/min for operation C. The 1-year graft survival rate was 100% and 89.5% in three " 5" donor group and 3-year graft survival rate was 100% and 84.2% respectively. Conclusions:Satisfactory outcomes might be obtained during dual kidney transplantation for infants and adults. The incidence of thrombosis, urine leakage and urinary protein is lower in improved kidney transplantation group than that in previously operated group. The problem of graft hyperperfusion injury is well solved. And the long-term follow-up outcome is excellent.

Objective:To observe the effect of transplanting neural stem cells (NSCs) and nerve growth factor (NGF) nanoparticles on hippocampal synaptophysin (SYN) levels in APP/PS1 double-transgenic mice and on their behavior.Methods:NSCs from the brains of transgenic mouse fetuses were cultured in vitro. Twenty-four 12-month-old male APP/PS1 mice with Alzhaimer′s disease (AD ) were randomly divided into an NSC combined with NGF transplantation group (NSCs+ NGF-NP group), an NSC transplantation group (NSCs group) and an AD control group (AD group), each of 8. Another 8 healthy male counterparts formed a normal control group (WT group). The rats in the NSCs+ NGF-NP and NSCs groups had NSCs combined with NGF nanoparticles or NSCs alone injected into their brains, while the bilateral hippocampi of the other two groups were injected with the same volume of phosphate buffer. Four weeks later the Morris water maze test was used to quantify the rats′ learning and memory functioning. The migration and differentiation of the transplanted cells were measured using fluorescence immunohistochemistry, while the level of synaptophysin protein in the hippocampus was detected using western blotting.Results:Neurospheres cultured in vitro showed positive expression of enhanced green fluorescent protein(EGFP) and nestin, which are specific markers of neural stem cells. Four weeks after the transplantation, cells traced using EGFP had survived and migrated to the corpus callosum and deep into the hippocampus and dentate gyrus. Transplanted cells had differentiated into DCX-positive neurons and GFAP-positive glial cells. The samples from the NSCs+ NGF-NP group showed a larger number of viable cells with longer protrusions, and some had crossed the hippocampal granular layer. The average synaptophysin levels in the WT, NSCs and NSCs+ NGF-NP groups were significantly higher than that of the AD group. The average synaptophysin level of the NSCs+ NGF-NP group was significantly higher than in the NSCs group but not significantly different from the WT group′s average. The Morris water maze results showed that compared with the AD group the time in the platform quadrant and the platform crossing times increased significantly in the WT, NSCs and NSCs+ NGF-NP groups. The average platform crossing time of the NSCs+ NGF-NP group was greater than that of the NSCs group but not significantly different from the WT group′s average.Conclusion:Injecting NSCs combined with NGF nanoparticles can promote the survival and maturation of the transplanted cells in vivo, increase hippocampal synapses, and thus improve learning and memory, at least in AD mice.

Different antiepileptic drugs (AEDs) may cause similar adverse effects, one of which is diplopia. However, the AEDs causing diplopia and the dose-response effect of each drug remains uncertain. In this study, we compared several second-generation AEDs to find out whether they would contribute to the risk of diplopia and their effect-causing dose. A meta-analysis was performed on 19 studies in agreement with our inclusion criteria. The results showed that eight commonly used second-generation AEDs (gabapentin, levetiracetam, oxcarbazepine, lamotrigine, pregabalin, topiramate, vigabatrin and zonisamide) could cause diplopia. The reported odds ratios (ORs) ranged from 1.406 to 7.996. Ranking risks from the highest to the lowest ORs of the eight AEDs of any dose resulted in the following order: use of oxcarbazepine (7.996), levetiracetam (7.472), lamotrigine (5.258), vigabatrin (3.562), pregabalin (3.048), topiramate (2.660), gabapentin (1.966), zonisamide (1.406). Taking into account the ORs above, we can conclude that second-generation AEDs of any dose may cause diplopia. However, the levetiracetam-caused diplopia needs to be further studied according to the data (OR, 7.472; 95% confidence interval, 0.375-148.772). These findings ask for better concerns about patients' quality of life when giving antiepileptic treatments.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Anticonvulsants ; therapeutic use ; Child ; Diplopia ; drug therapy ; Humans ; Middle Aged ; Placebo Effect ; Young Adult

Different antiepileptic drugs (AEDs) may cause similar adverse effects, one of which is diplopia. However, the AEDs causing diplopia and the dose-response effect of each drug remains uncertain. In this study, we compared several second-generation AEDs to find out whether they would contribute to the risk of diplopia and their effect-causing dose. A meta-analysis was performed on 19 studies in agreement with our inclusion criteria. The results showed that eight commonly used second-generation AEDs (gabapentin, levetiracetam, oxcarbazepine, lamotrigine, pregabalin, topiramate, vigabatrin and zonisamide) could cause diplopia. The reported odds ratios (ORs) ranged from 1.406 to 7.996. Ranking risks from the highest to the lowest ORs of the eight AEDs of any dose resulted in the following order: use of oxcarbazepine (7.996), levetiracetam (7.472), lamotrigine (5.258), vigabatrin (3.562), pregabalin (3.048), topiramate (2.660), gabapentin (1.966), zonisamide (1.406). Taking into account the ORs above, we can conclude that second-generation AEDs of any dose may cause diplopia. However, the levetiracetam-caused diplopia needs to be further studied according to the data (OR, 7.472; 95% confidence interval, 0.375-148.772). These findings ask for better concerns about patients' quality of life when giving antiepileptic treatments.

Objective To clinically analyze the frequent stroke localizations of moyamoya disease in order to improve our cognition toward it and reduce missed diagnosis. Methods All 32 patients were prospectively analyzed over the past 10 years in our hospital. Results The ratio of female to male was 1.28 and their age of onset ranged from 7 to 47 years old. Mean age of 5 ischemic stroke patients (15.6%) was 24 and mean age of 21 haemorrhagic stroke patients (65.6%) was 33 while mean age of 6 mixed type stroke patients (18. 8% ) was 32. The frequent ischemic stroke localizations were frontoparietal lobe (60%),temporo-occipital lobe (20%), and periventricular zone (20%). The frequent haemorrhagic stroke iocalizations were periventricular zone (42.8%), ventricle (39.2%), temporo-occipital lobe (10. 8%) and subarachnoid space (7.2%). No cerebellum and brain stem stroke occurred. Conclusion Adult patients usually develop intracranial hemorrhage. Moyamoya disease should be considered severely when adult patients develop periventricular and ventricular intracranial hemorrhage, frontoparietal cerebral infarction or adolescent patients develop ischemic stroke, especially companied with epilepsy.

Objective To investigate the dynamic changes of adenosine receptors, A1 (A1R) and A2a (A2aR) in the brain from the acute to chronic phase after kindling and to explore the correlation between seizure and expression level of A1R and A2aR. MethodsRats were randomly selected into the testing model, reference and normal control groups.Testing rats were kindled by lithium choride-pilocarpine, reference rats were treated with saline, and no treatment was given in normal control group. The dynamic expression of A1R and A2aR were detected by RT-PCR, immunofluorescence staining and Western blot at time-points of 24 hours, 1 month and 6 months post-kindling. Results In the acute phase of 24 hours after kindling, the A1R expression level (mRNA level was (1. 1483 ±0. 1182); Western blot result was ( 0. 7872± 0. 0621 ) ; immunofluorescence staining count was ( 76. 17 ± 4. 62 )/HP) was increased and A2aR (mRNA level was (0. 8338±0. 0572) ; Western blot result was (0. 2098 ±0. 0257) ; immunofluorescence staining count was (43. 83 ± 5. 12 )/HP) was decreased.The results showed statistically difference compared with the reference and normal groups (P< 0. 05 ). In the later chronic phase of 1 month and 6 months after kindling, the expression level of A1R was decreased and A2aR was increased. These data revealed statistically significant difference (P <0. 01 ). Furthermore, the comparison of the results in 1 month and 6 months after kindling found that the expression of AIR was lower in 6 months (mRNA level was (0. 5682 ±0. 0443) ; Western blot result was (0. 7749 ±0. 0262) ; immunofluorescence staining count was (38. 50 ±4. 81 )/HP) than in 1 month and that of A2aR was higher in 6 months (mRNA level was (1. 2169±0. 0332) ; Western blot result was (0. 7080 ±0. 0371 ); immunofluorescence staining count was (114. 50 ± 4. 04)/HP). The differences were statistical significant (t = - 19. 02--13.28, P < 0. 05). ConclusionsThe expressions of A1R and A2AR during and after kindling presents a bidirectional change. In the acute phas the expression of AR is regulated to suppress seizures. While in the chronic phase, the repeated seizures result in the change of A1R and A2aR expression in the opposite direction. This mechanism plays an important role in refractory seizures.

The clinical characters, diagnosis and differential diagnosis of paroxysmal kinesigenic choreoathetosis (PKC), and efficacy of the anti-epileptic drugs (AEDs) were investigated. Thirty-one patients with PKC were collected, and the clinical characters and change of EEG were analyzed. The average age of the first attack was 16.8 years old and the pinnacle was 10 to 20 years old. There were definite causes for every attack and the sudden movement was the most common one (92%). Time for the whole attack was always less than 1 min. The attack presented with muscle tension disturbance (83.9%), movement like dancing (16.1%), abnormal movement of mouth and face and other symptoms (16.2%). The attack tended to be very frequent and 71% patients were beyond once per day. The EEG examination and image scan of primary PKC were normal in most patients. Low dosage of AEDs could control the attack of 50%-77.3% patients. It was concluded that PKC was a common disease of movement disorder. The therapy by AEDs was very effective. PKC should be differentiated from epilepsy and the relationship between PKC and epilepsy needs further research.
Anticonvulsants/*therapeutic use ; Diagnosis, Differential ; Dystonia/diagnosis ; Dystonia/drug therapy ; Electroencephalography ; Epilepsy/diagnosis ; Young Adult