Alzheimer disease(AD),commonly known as senile dementia,is the most common type of dementia,resulting in progressive impairment of cognitive function,and is often accompanied by a variety of psychiatric symptoms,such as agitation.Agitated symptoms in AD patients often cause an increasing burden on caregivers,and current psychiatric medications may exacerbate adverse effects such as cognitive impairment and motor retardation in patients.Therefore,non-drug intervention is a very important adjuvant treatment option.This article reviews the clinical manifestations,possible mechanisms,drug therapy and non-drug intervention measures of agitation in order to provide reference for more effective treatment of AD.

Objective:To explore the clinical characteristics of pediatric kidney transplant recipients reinfected with SARS-CoV-2.Method:The relevant clinical data were retrospectively reviewed for 191 pediatric kidney transplant recipients at a single center. Based upon whether or not there was a reinfection of SARS-CoV-2, they were assigned into two groups of single infection (group A, 127 cases) and reinfection (group B, 64 cases). Baseline profiles, clinical symptoms, diagnostic and therapeutic strategies, markers of disease progression, immune status, respiratory support modalities, comorbidities and transplantation-related data were collected for comparing the inter-group differences during primary infection and between two infections in reinfected group.Result:As compared with group A, group B recipients had a higher proportion of age <12 years (71.9% vs 54.3%) ,unvaccinated (81.2% vs 66.1%) and such symptoms as high fever (34.4% vs 12.6% ), dry cough (43.8% vs 23.6% ) and chest tightness (14.1% vs 3.9 %) during primary infection (all P<0.05). During primary infection, the levels of IL-6 and CRP were higher in group B than in group A and inter-group difference was statistically significant (both P<0.01). The levels of IL-6 ( P<0.01), CRP ( P<0.01) and PCT ( P= 0.023) were lower in group B during reinfection than those during primary infection and the difference was statistically significant. During primary infection, the counts of CD3+, CD4+, CD8+, NK and B lymphocyte of group B were lower than those of group A. And inter-group differences were statistically significant (all P<0.01). During reinfection, the levels of CD3+, CD4+, CD8+, NK and B lymphocyte counts of group B spiked as compared with those of group A during primary infection and the differences were statistically significant (all P<0.01). The levels of SCr and UA in group B differed insignificantly before and after primary infection with SARS-CoV-2. However, the differences before and after reinfection were statistically significant (both P<0.01) . Conclusion:Symptomatic and immunocompromised pediatric KT recipients during primary infection with SARS-CoV-2 are more prone to reinfection during subsequent epidemics. Though mildly symptomatic, reinfection may exacerbate impairments of graft kidney function in pediatric KT recipients.

Ultra-rapid cooling and rewarming rate is a critical technical approach to achieve ice-free cells during the freezing and melting process. A set of ultra-rapid solid surface freeze-thaw visualization system was developed based on a sapphire flim, and experiments on droplet freeze-thaw were carried out under different cryoprotectant components, volumes and laser energies. The results showed that the cooling rate of 1 μL mixed cryoprotectant [1.5 mol/L propylene glycol (PG) + 1.5 mol/L ethylene glycol (EG) + 0.5 mol/L trehalose (TRE)] could be 9.2×10 ³ °C/min. The volume range of 1-8 μL droplets could be vitrified. After comparing the proportions of multiple cryoprotectants, the combination of equal proportion mixed permeability protectant and trehalose had the best vitrification freezing effect and more uniform crystallization characteristics. During the rewarming operation, the heating curve of glassy droplets containing gold nanoparticles was measured for the first time under the action of 400-1 200 W laser power, and the rewarming rate was up to the order of 10 ⁶ °C/min. According to the droplet images of different power rewarming processes, the laser power range for ice-free rewarming with micron-level resolution was clarified to be 1 400-1 600 W. The work of this paper simultaneously realizes the ultra-high-speed temperature ramp-up, transient visual observation and temperature measurement of droplets, providing technical means for judging the ice free droplets during the freeze-thaw process. It is conducive to promoting the development of ultra-rapid freeze-thaw technology for biological cells and tissues.
Freezing ; Vitrification ; Cryopreservation/methods* ; Trehalose ; Gold ; Rewarming ; Metal Nanoparticles ; Cryoprotective Agents ; Lasers

Objective:To investigate the short-term efficacy and safety of Donafenib as postoperative adjuvant therapy for patients with high risk of recurrence after radical resection of hepatocellular carcinoma (HCC).Methods:The propensity score matching (PSM) and retrospective cohort study was conducted. The clinicopathological data of 157 HCC patients with high risk of recurrence after radical resection who were admitted to 6 medical centers, including The First Affiliated Hospital of Nanjing Medical University et al, from June 2021 to February 2023 were collected. There were 128 males and 29 females, aged (59±10)years. Of 157 patients, 101 cases undergoing Donafenib as postoperative adjuvant therapy were divided into the the Donafenib group, and 56 cases under-going no systemic postoperative adjuvant therapy were divided into the control group. Observation indicators: (1) PSM and comparison of general data of patients between the two groups after matching; (2) postoperative treatment; (3) follow-up and survival of patients; (4) analysis of risk factors affecting recurrence-free survival of patients. PSM was done based on the principle of optimal perfect matching, with the clamp value of 0.5, and the Donafenib group and the control group were matched at a ratio of 1.25∶1. Measurement data with normal distribution were represented as Mean± SD, and comparison between groups was conducted using the t test. Measurement data with skewed distribution were represented as M(range). Count data were described as absolute numbers and/or percentages, and comparison between groups was conducted using the chi-square test. Comparison of ordinal data between groups was conducted using the Kruskal-Wallis H test. The Kaplan-Meier method was used to calculate survival rates and draw survival curves, and the Log-Rank test was used for survival analysis. The COX proportional hazard model was used for univariate and multivariate analyses. Results:(1) PSM and comparison of general data of patients between the two groups after matching. Of 157 patients, 126 cases were successfully matched, including 70 cases in the Donafenib group and 56 cases in the control group, respectively. The elimination of tumor number confounding bias ensured comparability between the two groups after PSM. (2) Postoperative treatment. After PSM, of 70 patients in the Donafenib group, there were 23 cases receiving Donafenib monotherapy, 26 cases combined with transcatheter arterial chemoembolization (TACE), 14 cases combined with immunotherapy, and 7 cases combined with TACE+immunotherapy. Of 56 patients in the control group, there were 37 cases receiving postoperative follow-up alone and 19 cases combined with TACE. (3) Follow-up and survival of patients. All 157 patients were followed up, and the follow-up time of the 101 patients in Donafenib group and the 56 patients in control group were 10.1(range, 6.3-14.6)months and 22.2(range, 15.1-25.5)months, respectively. During the follow-up period, 70 patients in the Donafenib group experienced treatment-related adverse reactions, inclu-ding 8 cases of grade 3 adverse reactions, 23 cases of grade 2 and 39 cases of grade 1 adverse reactions, respectively. After PSM, the postoperative 12-, 18-month recurrence-free survival rates were 83.7%, 83.7% in the 70 patients of Donafenib group and 67.8%, 58.9% in the 56 patients of control group, respectively, showing a significant difference in the postoperative recurrence-free survival time between the two groups ( hazard ratio=0.395, 95% confidence interval as 0.176-0.888, P<0.05). (4) Analysis of risk factors affecting recurrence free survival of patients. Results of multivariate ana-lysis showed that microvascular invasion, vascular thrombus, clinical stage as ⅢA were independent risk factors affecting recurrence-free survival in patients with high risk of recurrence after radical resection of HCC ( hazard ratio=2.181, 2.612, 2.612, 95% confidence interval as 1.028-4.629, 1.128-6.047, 1.128-6.047, P<0.05), Donafenib as postoperative adjuvant therapy was an independent protective factor affecting recurrence-free survival in patients with high risk of recurrence after radical resection of HCC ( hazard ratio=0.457, 95% confidence interval as 0.227-0.920, P<0.05). Results of further analysis showed that after PSM, there were significant differences in the postoperative recurrence-free survival time in patients with different clinical factors, including male, age ≥60 years, tumor diameter >5 cm, positive microvascular invasion, positive hepatitis B virus infection, alpha fetoprotein <200 μg/L, between the Donafenib group and the control group ( hazard ratio=0.283, 0.202, 0.174, 0.345, 0.273, 0.180, 95% confidence interval as 0.114-0.707, 0.044-0.937, 0.038-0.794, 0.128-0.929, 0.091-0.819, 0.052-0.620, P<0.05). Conclusion:Donafenib as postoperative adjuvant therapy can effectively reduce the short-term recurrence rate in patients with high risk of recurrence after radical resection of HCC, with good safety and tolerance.

Objective:To investigate the additional value of unenhanced computed tomography (CT) in the differential diagnosis of brain tumors and non-neoplastic lesions.Methods:A total of 237 cases [140 males and 97 females; (49±16) years old; including 48 cases of low-grade glioma, 134 cases of high-grade glioma, 38 cases of primary central nervous system lymphoma, 9 cases of medulloblastoma, 5 cases of germinoma, and 3 cases of central neurocytoma] of brain tumors (diffuse gliomas and non-glial tumors) diagnosed by biopsy or surgery and pathology in the Affiliated Hospital of Qingdao University from September 2016 to October 2020 were collected retrospectively. Sixty-six cases [46 males and 20 females; (42±13) years old; including 12 cases of abscesses, 5 cases of infarcts, 33 cases of demyelinating lesions, 11 cases of autoimmune encephalitis, and 5 cases of central nervous system vasculitis] of brain non-neoplastic lesions were confirmed by biopsy or clinic. All patients underwent routine magnetic resonance imaging (MRI) scan and unenhanced CT before the treatment. The images were reviewed by two neuroradiologists together blind to the final diagnosis with and without CT images respectively. The diagnostic results and reliability scores were recorded, and the accuracy of the two evaluations was compared.Results:CT hyperattenuation exhibited a higher specificity (95%) than conventional MRI scan (86%), and a lower diagnostic sensitivity (34% vs 86%). Compared to MRI alone, the combined modality of MRI and unenhanced CT significantly improved diagnostic accuracy (94% vs 86%). Additionally, the CT attenuation ratio of non-neoplastic lesions was significantly lower than that of neoplastic lesions [0.69 (0.61,0.78) and 1.14 (1.00,1.25), W=2 123, P<0.05]. The CT attenuation ratio in the non-glial origin tumor group was significantly higher than that in the diffuse glioma group [1.28 (1.18,1.41) and 1.13 (0.97,1.21), W=1 858, P<0.05]. There was no significant difference in grade Ⅲ and Ⅳ groups of diffuse glioma [1.11 (0.99,1.20) vs 1.16 (1.09,1.24), P>0.05 (Nemenyi test)]. However, both were significantly higher than that of grade Ⅱgroup of diffuse glioma [0.89 (0.76,1.07), P<0.05 (Nemenyi test)]. No significant difference was observed between astrocytic tumors and oligodendroglial tumors at the same grade. Conclusions:Hyperattenuation on unenhanced CT is highly specific for the diagnosis of brain tumors. Unenhanced CT plus MRI is more accurate for distinguishing the two entities in hypoattenuation lesion on unenhanced CT.

Objective:To evaluate the effect of unilateral pediatric kidney donation for adult kidney transplantation.Methods:Retrospective analysis was conducted on the cases of children who donated unilateral donor kidney for adult kidney transplantation recipients in our hospital, and those who were followed up for more than three years were included in this study. The body weight of the recipients in group A was ≤50 kg, and the body weight of the recipients in group B was ≤70 kg.The recipients were divided into 0-5 year old donor group (group A) and 6-17 year old donor group (B group). Clinical data, recipient/kidney survival, graft function and growth, and complications of the recipient were analyzed.Results:A total of 45 adult recipients were enrolled, including 12 in group A and 33 in group B. The renal survival rate at 3 years after operation was (100%, 96.9%)/(91.6%, 93.9%). One week after the operation, the early postoperative recovery of renal function in group B was better than that in group A, and the difference of serum creatinine was statistically significant ( P<0.05), while the difference of serum creatinine in other postoperative follow-up time points was not statistically significant ( P>0.05). Within a year, both groups of grafts continued to grow, reaching adult levels in one year. There was no statistical significance in the incidence of complications between the two groups ( P>0.05). The incidence of protein in the two groups was 33.3% and 6.1%, respectively, 1 case in each group still had proteinuria at 1 year after surgery, and only 1 case in the infant donor kidney recipient in group A had proteinuria at 3 years after surgery. Conclusions:Unilateral donor kidney transplantation from children can provide good results for adult patients with uremia by selecting suitable donors according to the weight of the recipient.

Objective:To explore the effect of drug combined with CO2 laser minimally invasive surgery in the treatment of patients with vascularized vocal cord inflammation, and the influence on the voice function of patients.Methods:From April 2018 to July 2019, 74 patients with vascularized vocal cord inflammation admitted in the Affiliated Hospital of Yangzhou University were divided into two groups according to the random number table method, with 37 cases in each group.The patients in the control group were treated with oral medicine.The patients in the observation group were treated with medicine combined with CO 2 laser minimally invasive surgery.The voice quality, improvement of voice disorders, recurrence and patients' satisfaction were observed. Results:After treatment for 12 months, the total hoarseness(G), roughness(R) and breath(B) scores in the observation group were (1.60±0.13)points, (1.71±0.21)points and (1.42±0.12)points, respectively, which were significantly lower than those in the control group [(1.88±0.16)points, (1.98±0.10)points and (1.78±0.13)points] ( t=8.262, 7.061, 12.377, all P<0.05). After treatment, the functional, physiological and emotional scores of voice disorders index(VHI) and total score in the observation group were (10.10±1.03)points, (13.26±2.10)points, (7.30±1.10)points, (31.26±4.74)points, respectively, which were significantly lower than those in the control group [(11.63±2.16)points, (15.42±2.16)points, (8.86±1.05)points, (35.86±5.33)points] ( t=3.889, 4.361, 6.240, 19.103, all P<0.05). The recurrence rates at the first and third month after treatment in the observation group were 2.70%(1/37) and 8.11%(2/37), respectively, which were significantly lower than those in the control group [16.22%(6/37) and 27.03%(10/37)] (χ 2=3.945, 4.573, all P<0.05). The satisfaction rate of patients in the observation group was 94.59%(35/37), which was significantly higher than 78.38%(29/37) in the control group(χ 2=4.163, P<0.05). Conclusion:On the basis of conventional drug treatment, CO 2 laser microinvasive surgery can effectively improve the treatment effect, improve the voice function of patients, and significantly reduce the recurrence rate, the overall effect and prognosis are better.

Objective:To determine the expression of microRNA-188-5p (miR-188-5p) in cutaneous squamous cell carcinoma (CSCC) tissues and cells, and to assess the effect of its downregulation on the proliferation and invasion of CSCC cells.Methods:From November 2012 to October 2018, 50 surgically resected CSCC tissue specimens and 50 paracancerous normal skin tissue specimens were collected from the First Affiliated Hospital of Xinxiang Medical College in Henan Province. Real-time fluorescence-based quantitative PCR (qPCR) was employed to determine the expression of miR-188-5p in CSCC tissues, paracancerous normal skin tissues, CSCC cell lines SCL-1, A431 and HSC-5, and a human immortalized keratinocyte line HaCaT. Cultured A431 and HSC-5 cells were both divided into 2 groups: miR-188-5p inhibitor group and negative control group, which were transfected with a miR-188-5p inhibitor and its negative control respectively. Then, qPCR was performed to determine the relative expression level of miR-188-5p (expressed as 2 -△△Ct), and cell counting kit-8 (CCK8) and Transwell assays were conducted to assess cellular proliferative activity and invasive ability respectively in the above groups. Dual-luciferase reporter assay was performed to investigate interactions between miR-188-5p and phosphatase and tensin homologue deleted on chromosome 10 (PTEN), and Western blot analysis to determine the protein expression of PTEN, total Akt (t-Akt) and phosphorylated Akt (p-Akt). Two independent samples were compared by using t test. Results:The relative expression level of miR-188-5p was significantly higher in the CSCC tissues (5.213 ± 3.138) than in the paracancerous normal skin tissues (1.010 ± 0.364, t = 9.187, P < 0.001), and significantly higher in the SCL-1, A431 and HSC-5 cells (3.858 ± 0.163, 7.068 ± 0.262 and 4.572 ± 0.413, respectively) than in the HaCaT cells (1.079 ± 0.300, t = 17.890, 21.110 and 8.737, respectively, all P < 0.05). Compared with the negative control group, the miR-188-5p inhibitor group showed significantly decreased miR-188-5p expression in both A431 and HSC-5 cells (both P < 0.01), and decreased proliferative activity and invasive ability of both A431 and HSC-5 cells (all P < 0.05). Dual-luciferase reporter assay showed that the downregulation of miR-188-5p significantly increased the expression of PTEN, but inhibited the expression of p-Akt in A431 and HSC-5 cells. Conclusion:MiR-188-5p is highly expressed in CSCC tissues and cells, and the downregulation of miR-188-5p may inhibit the proliferative activity and invasive ability of CSCC cells by regulating the PTEN/Akt pathway.

Objective To evaluate the effect of downregulation of microRNA (miR)-373 expression on cell cycle and apoptosis of a cutaneous squamous cell carcinoma (CSCC) cell line A431.Methods A431 cells at exponential growth phase were classified into 3 groups:miR-373 inhibitor group and negative control group transfected with miR-373 inhibitor and negative control miRNA respectively,and untreated group receiving no treatment.At 48 hours after the transfection,real-time PCR was performed to determine the expression of miR-373 in the above 3 groups,cell counting kit-8 (CCK-8) assay to evaluate the effect of downregulated expression of miR-373 on the proliferation of A431 cells,flow cytometry to investigate the distribution of cell cycle and changes in apoptosis of A431 cells in different treatment groups,and colorimetric analysis to detect the changes in caspase-3 activity in different treatment groups.Statistical analysis was carried out with SPSS 17.0 software by using two-sample t test for the comparison between two groups,one-way analysis of variance (ANOVA) for the comparison among 3 groups,and least significant difference (LSD)-t test for multiple comparisons.Results The expression of miR-373 was significantly lower in the miR-373 inhibitor group (0.120 ± 0.036) than in the untreated group (1.002 ± 0.022) and negative control group (1.037 ± 0.028,LSD-t =36.21,34.83,respectively,both P ＜ 0.001).At 48,72 and 96 hours,the miR-373 inhibitor group showed significantly decreased proliferative activity of A375 cells compared with the untreated group and negative control group (F =10.805,13.720 and 30.907 respectively,P =0.038,0.010 and 0.001 respectively).The proportion of A375 cells in G0/G1 phase was significantly higher in the miR-373 inhibitor group (64.69％ ± 1.18％) than in the untreated group (52.74％ ± 0.66％,t =15.51,P ＜ 0.001) and negative control group (53.80％ ± 0.80％,t =13.24,P ＜ 0.001).The proportion of total apoptotic cells and activity of caspase-3 in the miR-373 inhibitor group were 22.69％ ± 1.24％ and 1.238 ± 0.057 respectively,which were significantly higher than those in the untreated group (9.62％ ± 1.14％,0.413 ± 0.028 respectively,both P ＜ 0.001)and negative control group (9.66％ ± 0.97％,0.437 ± 0.036 respectively,both P ＜ 0.001).Conclusion MiR-373 may play an important role in the regulation of cell cycle and induction of apoptosis of the CSCC cell line A431.

Objective: To explore the reliability, validity and best cut-off value of Chinese version Addenbrooke's cognitive examination-Ⅲ(ACE-Ⅲ) in presbycusis patients with mild cognitive impairment(MCI). Methods: A total of 43 presbycusis patients with mild cognitive impairment, 61 presbycusis patients with normal cognitive impairment and 60 healthy controls treated in outpatient clinic of department of otolaryngology in the Affiliated Hospital of Yangzhou University from January 2016 to October 2017 were enrolled.The Chinese version of mini mental state examination(MMSE), Montreal cognitive assessment(MoCA) and ACE-Ⅲ were used to evaluate cognitive function of subjects.The reliability and validity of the ACE-Ⅲ were analyzed with the SPSS 21.0, and the cut-off point was confirmed with the receiver operating characteristic(ROC) curve analysis, and the value of the scale questionnaire in the diagnosis of presbycusis patients with MCI was assessed. Results: Chinese version of ACE-Ⅲ had betler feasiblity, and the receiving rate, recovery rate and effivience were 100.0%, 100.0% and 94.9%, respetively.The Cronbach's alpha, split-half reliability and intraclass correlation coefficient were 0.870(P<0.05), 0.874(P<0.05) and 0.880(P<0.05), respectively.The patients in MCI group got a lower score in the sub-scale of attention/orientation, memory, verbal fluency, language and visual space of ACE-Ⅲ compared with those in NCI group and healthy control group, and the differences were statistically significant (F=48.042, 46.594, 35.442, 19.374, 256.19, 140.319, all P<0.001). The criteria validity calculated between the Chinese version of ACE-Ⅲ and MMSE was 0.802 (P<0.001). Factor analysis of the construct validity showed that the 26 items were classed into five domains.Every domain was sensitive and effective to discriminate between patients and healthy individuals(P<0.05). The Chinese version of ACE-Ⅲ showed significantly correlation with the total scores of MoCA(r=0.802, P<0, 001). When the cut-off value for diagnosis was 86.5, the largest area under the ROC curve for the Chinese version of ACE-Ⅲ was 0.98(95% CI: 0.897-0.996). The relative sensitivity and specificity were 100.0% and 95.0%, respectively. Conclusion The Chinese version of ACE-Ⅲ had better internal consistency, reliability and validity, and is applicable for MCI assessment in elderly patients with presbycusis.Patients with presbycusis can be considered to have cognitive dysfunction with score of 86.5 by using this questionnaire.