Objective:To analyze the growth characteristics of different genotypes of Japanese encephalitis virus (JEV) in different cell lines, and to provide scientific basis for the selection of cell lines in the study of JEV.Methods:BHK-21, Vero, C6/36, PK-15, DF-1, N2a, SH-sy5y and MDCK cell lines were selected. The proliferation ability of genotype 1 (NX1889 strain), genotype 3 (P3 strain) and genotype 5 (XZ0934 strain) JEV in these cell lines was evaluated by plaque assay and RT-qPCR.Results:Significant cytopathogenic effects (CPE) were observed in BHK-21, Vero, C6/36, DF-1, N2a and PK-15 cell lines across all three JEV genotypes. However, no significant differences in CPE characteristics were observed within the same cell line. SH-sy5y and MDCK cell lines did not show significant CPE, but virus proliferation was detected in SH-sy5y cell line, while MDCK cell line were found to be insensitive to JEV. No significant difference was observed in the proliferation curves of G1, G3 and G5 JEV in BHK-21, Vero and SH-sy5y cell lines. In C6/36 and PK-15 cell lines, the titer of G1 JEV was higher than that of G3 and G5. In DF-1 cell line, G5 demonstrated a higher titer than the other two genotypes, whereas in N2a cell line, G5 showed a lower titer than the other two.Conclusions:There are differences in the proliferation of three different genotypes of JEV in different cell lines, which can provide reference for the study of JEV in different directions.

Optogenetic technique is a cellular activity regulation technology that combines optics and genetics, which can specifically regulate the relevant brain regions and neural circuits in depressive animal models, thereby slowing down or aggravating depression-like behaviors in experimental animals. Optogenetic technique combined with neuroimmunoassay, behavioral detection and brain imaging techniques can provide technical support for elucidating the pathogenesis of depression, developing new antidepressants and diagnosis and treatment methods. In this paper, the application of optogenetic technique in the ventral tegmental area, nucleus accumbens, prefrontal cortex, hippocampus, dorsal raphe nucleus and other brain regions closely related to depression is reviewed to provide ideas and directions for study of neural circuits of depression.

Objective:To compare the effect of neoadjuvant chemotherapy vs. concurrent chemoradiotherapy on the target volume and organs at risk for locally advanced bulky (>4 cm) cervical cancer. Methods:From March 1, 2019 to June 30, 2021, 146 patients pathologically diagnosed with cervical cancer were selected and randomly divided into two groups using random number table method: the neoadjuvant chemotherapy (NACT) + concurrent chemoradiotherapy (CCRT) group ( n=73) and CCRT group ( n=73). Patients in the NACT+CCRT group received 2 cycles of paclitaxel combined with cisplatin NACT, followed by CCRT, the chemotherapy regimen was the same as NACT. In the CCRT group, CCRT was given. Statistical description of categorical data was expressed by rate. The measurement data between two groups were compared by Wilcoxon rank-sum test for comparison of two independent samples, and the rate or composition ratio of two groups was compared by χ2 test. Results:Before radiotherapy, GTV in the NACT+CCRT group was (31.95±25.96) cm 3, significantly lower than (71.54±33.59) cm 3 in the CCRT group ( P<0.01). Besides, CTV and PTV in the NACT+CCRT group were also significantly lower compared with those in the CCRT group (both P<0.05). In terms of target volume dosimetry, D 100GTV, D 95CTV, V 100GTV, V 100CTV and V 95PTV in the NACT+CCRT group were significantly higher than those in the CCRT group (all P<0.05). The complete remision (CR) rates in the NACT+CCRT and CCRT groups were 86.3% and 67.6%, with statistical significance between two groups ( P<0.01) . Regarding organs at risk, NACT+CCRT group significantly reduced the dose to the bladder, rectum, small intestine and urethra compared with CCRT group (all P<0.05). Conclusions:NACT can reduce the volume of tumors in patients with large cervical masses, increase the radiation dose to tumors, reduce the dose to organs at risk, and make the three-dimensional brachytherapy easier. Therefore, NACT combined with CCRT may be a new choice for patients with locally advanced cervical cancer with large masses.

Objective:To investigate the optimal bladder filling volume in the 3D brachytherapy of postoperative cervical cancer.Methods:Totally 111 early cervical cancer patients with positive incisal margins or insufficient safety boundaries were included. The normal saline 50, 60, 70, 80, 90, and 100 ml were filled into their bladders, and accordingly six groups were determined, and 66, 69, 66, 69, 72, 56 person-times in each group, respectively. The CT-based simulation positioning was performed. According to the ICRU 89 report, high-risk clinical target volume and organs at risk such as bladder and rectum were delineated. The Oncentra planning system was used to prepare the treatment program. The high-risk clinical target volume (HR-CTV), D90, and the D2 cm 3 and D1 cm 3 of organs at risk under different volumes were recorded. Results:Compared to the 60 ml group, the volume and dosage of HR-CTV in the groups of 50, 70, 80, 90, and 100 ml had no significant difference ( P>0.05). The D2 cm 3 and D1 cm 3 of the bladder and rectum of patients in these groups significantly decreased, and the difference was statistically significant ( tbladder = 3.21, 5.83, 2.89, 12.95, 7.96, Pbladder = 0.031, 0.010, 0.041, 0.000, 0.001; trectum = 2.94, 4.66, 2.53, 5.89, 4.13, Prectum = 0.037, 0.024, 0.049, 0.005, 0.028). The pairwise comparison among these groups except for the 60 ml group showed that the volume and dosage of HR-CTV and the D2 cm 3 and D1 cm 3 of bladder and rectum had no significant difference ( P > 0.05). Moreover, the D2 cm 3 and D1 cm 3 of sigmoid colon and small intestine of these groups had no significant difference ( P > 0.05). Conclusions:In the 3D brachytherapy of postoperative early cervical cancer, a bladder filling volume of 60 ml can ensure the volume and dose of HR-CTV and can protect the bladder and rectum compared with other filling volumes.

Objective:To explore the clinical features of three early-onset infantile epileptic encephalopathy (EIEE) patients with variations in phosphofurin acidic cluster sorting protein 2 (PACS2) gene and to review related literature.Methods:The clinical data and genetic features of three early infantile epileptic encephalopathy 66 (EIEE66) patients with a PACS2 gene variant diagnosed by the Department of Neurology, Wuhan Children′s Hospital, Tongji Medical College, Huazhong University of Science and Technology, from January 2019 to January 2020 were retrospectively analyzed. A literature search with "PACS2 gene" "PACS2" "epileptic encephalopathy, early infantile, 66" and"early infantile epileptic encephalopathy 66" as key words was conducted at PubMed, China National Knowledge Infrastructure (CNKI), and Wanfang Data Knowledge Service Platform (up to July 2020). Case reports of patients with PACS2 gene variants and related clinical data were chosen and reviewed.Results:Case 1, a girl aged 2 years and 2 months was hospitalized because of repetitive seizures within more than two years and 6 convulsions within 2 days due to fever. The seizures occurred at the age of 7 days, characterized by focal seizures and generalized tonic-clonic seizures. Sometimes, the frequency of seizures increased with high fever. Regular treatment had not been implemented in the early stage, later seizures were controlled by valproic acid treatment. Case 2, a female 5 months of age, was admitted due to recurrent convulsions in nearly five months. Focal seizures occured at the age of 5 days. And the brain magnetic resonance imaging (MRI) confirmed abnormal cerebellar hemispheres and cerebellar vermis, as well as cerebellar dysplasia. Several antiepileptic drugs and ketogenic diet were ineffective in the early months, and later seizures were controlled with the treatment with levetiracetam and valproic acid. Case 3, a five-month-old girl, was admitted because of recurrent convulsions for nearly five months. At the age of 3 days, she had tonic seizures, and showed good response to levetiracetam and valproic acid. All the three cases were accompanied by development delay and dysmorphic facial appearance, and got seizure-free with the treatment with valproic acid. All copy-number variant analysis and trio whole exome sequencing revealed a recurrent heterozygous missense variant (c.625G>A) in PACS2 gene. No related reports were found in Chinese journals, while 4 reports were found in English literature, describing 17 patients in total. With these 3 patients included, 20 cases had only two missense PACS2 gene variants, in whom 19 cases carried the variant c. 625G>A (p.Glu209Lys) and 1 case carried the variant c. 631G>A (p.Glu211Lys). Epilepsy was the first reported symptom in all patients, and 17 cases had seizures during the first week of life. Out of the various seizure types observed, focal seizures were the predominant types (13 cases), whereas tonic, clonic, tonic-clonic seizures and non-motor seizures (such as facial flushing) were also reported. Almost all patients showed facial dysmorphism and developmental delay to different degrees. Total of 16 patients had abnormal brain MRI recordings, and 13 cases had cerebellar hypoplasia. More specifically, 7 cases showed inferior vermian hypoplasia, and 3 cases showed hypothalamic fusion anomaly. The treatment was mainly aimed to control the symptoms. And the recommended effective treatment for epilepsy has not been reported yet.Conclusions:PACS2-related early infantile epileptic encephalopathy is an autosomal dominant disease, characterized by seizure onset within the first week of life in most cases, dysmorphic facial appearance, and various degrees of developmental retardation. Treatment with valproic acid showed good effect.

Objective:Exploring tau related disease pattern (tauRDP) in the brain of Alzheimer′s disease (AD) patients based on 18F-APN-1607 PET scan. Methods:18F-APN-1607 PET images were collected from 17 AD patients (6 males and 11 females, age: (61.7±12.3) years, Mini-Mental State Examination (MMSE) score: 17.6±7.9) and 10 normal controls (NC; 6 males and 4 females, age: (61.2±4.7) years) from Huashan Hospital of Fudan University. The scaled subprofile model (SSM) based on principal component analysis (PCA) technique was used to construct the tauRDP. Then the expression value of tauRDP in each sample was calculated. The differences on tauRDP expression values between AD patients and NC were compared by independent-sample t test. Pearson correlation analysis was used to analyze the correlation between tauRDP expression values and MMSE values in AD patients. Results:The tauRDP area mainly included: precentral gyrus, dorsolateral superior frontal gyrus, middle frontal gyrus, inferior frontal gyrus of opercular part, inferior frontal gyrus of triangular part, supplementary motor area, medial superior frontal gyrus, left median cingulate and paracingulate gyri, right cuneus, superior occipital gyrus, middle occipital gyrus, postcentral gyrus, superior parietal gyrus inferior parietal, but supramarginal and angular gyri, supramarginal gyrus, angular gyrus, precuneus and middle temporal gyrus. There were significant differences ( t=4.395, P<0.001) between AD group (12.6±8.0) and NC group (0.0±1.0) in tauRDP expression values. The tauRDP expression values were correlated with MMSE values in AD group significantly ( r=-0.566, P=0.018). Conclusions:TauRDP established basing on SSM/PCA method can be used to quantitatively express the abnormal spatial distributions of tau deposition. Expression value of tauRDP has the potential to initially assess the severity of AD.

Objective To explore the prognostic factors for the recurrence of vaginal stump of earlystage cervical cancer after radical hysterectomy and evaluate the effect on clinical prognosis.Methods Clinical data of stage Ⅰ-ⅡA cervical cancer patients undergoing radical hysterectomy in Guizhou Cancer Hospital from January,2007 to December,2016 were retrospectively analyzed.Results A total of 493 patients were enrolled and followed up until May 30,2018.Among them,96.6％(474/493) completed the follow-up.The median age was 45 years.Patients aged 40-50 years had a high incidence rate.In total,451 cases (91.48％) had no recurrence of vaginal stump.The average time without stump recurrence was 51.2 months and the median time without stump recurrence was 44.8 months.Multivariate Cox regression analysis demonstrated that pelvic external irradiation and brachytherapy were the independent prognostic factors for the recurrence of vaginal stump (P=0.000,0.000).Tumor size,lymph node metastasis and pelvic external irradiation were the independent prognostic factors for overall survival (P=0.045,0.022,0.000).Conclusions Pelvic external irradiation and brachytherapy play an extremely pivotal role in reducing the risk of vaginal stump recurrence after radical hysterectomy for patients with stage Ⅰ-Ⅱ A cervical cancer.Tumor size,lymph node metastasis and pelvic external irradiation are the independent prognostic factors for overall survival of patients with stage Ⅰ-Ⅱ A cervical cancer following radical hysterectomy.

Objective To compare the effect of different therapeutic methods upon the survival of stage Ⅰ-Ⅱ A cervical cancer patients with intermediate risk factors and explore the optimal treatment for patients with early-stage cervical cancer undergoing radical hysterectomy and pelvic lymphadenectomy.Methods Clinical data of 323 patients with the following intermediate risk factors of lymphovascular space invasion,depth of stromal invasion or tumor size ＞ 4 cm were retrospectively analyzed.The impact of observing (NT),chemotherapy (CT),radiotherapy (RT) and concurrent chemoradiotherapy (CCRT) on survival was statistically compared.The Kaplan-Meier method was used to survival analysis,and log-rank test difference,Cox model was used to prognostic factor analysis.Results The 5-year progression-free survival (PFS) and overall survival (OS) of all patients were 79.0％ and 84.8％.Univariate and multivariate analyses demonstrated that TS＞ 4 cm and therapeutic method were the independent prognostic factors of PFS.The number of risk factors and therapeutic method were the independent prognostic factors of OS.In the whole group,both RT and CCRT could improve the prognosis of patients with no statistical significance (P＞0.05).In the subgroup analysis,for patients with a single intermediate risk factor (low risk group),CT could significantly prolong the PFS (P=0.026) rather the 5-year OS (P=0.692).Compared with NT and CT,RT and CCRT could improve the PFS and OS,whereas no statistical significance was noted between the RT and CCRT (both P＞0.05).For those with ≥2 risk factors (high risk group),CCRT could significantly prolong the PFS compared with CT (84.9％ vs.70％;P=0.006),but did not improve the OS (P=0.107).Compared with RT,CCRT could significantly improve the PFS and OS (both P＜0.05).Conclusion For patients with only one risk factor,RT can enhance the clinical prognosis.CCRT can improve the clinical prognosis of stage Ⅰ-Ⅱ A cervical cancer patients with ≥ 2 risk factors.

Extra-articular distal tibial fractures as a result of high-energy damage are often comminuted or displaced, frequently accompanied by severe soft tissue injuries.Poor blood supply and various complications make the treatment more difficult,affecting life quality of the patients.The main goals of the treatment are to abtain a healed,well-aligned fracture,functional range of motion of the ankle joint and minimizing complications.It is generally recommended that surgical treatment be performed in the proper context of local conditions to facilitate early functional exercise.Plate fixation and intramedullary nail fixation are the common options for closed fractures.This article focuses on the two treatment methods and some important auxiliary technologies in both domestic and foreign, hoping to provide some references for clinical treatment.

Objective: To observe the effects of miR-204 on the proliferation and invasion of retinoblastoma (RB) cells and to explore the potential regulatory mechanism. Methods: The expression level of miR-204 in RB cell lines (Y79, SO-RB50, and HXO-Rb44) as well as in normal human retinal pigment epithelial cell line hTERT RPE-1 was detected using qRT-PCR. The Y79 cells were divided into two groups (negative control group and miR-204 group) by respectively transfecting Y79 cells with NC-mimics and miR-204 mimics using liposome transfection method. The effects of miR-204 on Y79 cell proliferation was detected with CCK-8 assay; while the effect of miR-204 on migration and invasion of Y79cellsweredeterminedbycellscratchassayandTranswellassay,respectively.Besides, thepotentialtargetgeneofmiR-204waspredictedbybioinformatics;and the influence of miR-204 on the expression of high mobility group AT-hook 2 gene (HMGA2) at both mRNA and protein levels was detected using qRT-PCR and Western blotting, respectively. Results: miR-204 expression in RB cell lines Y79, SO-RB50 and HXO-Rb44 was remarkably lower than that in normal human retinal pigment epithelial cell line hTERT RPE-1 (P<0.01). miR-204 expression in Y79 cells was markedly up-regulated after transfection with miR-204 mimics (P<0.01) along with significantly reduced cell proliferation, migration and invasion capacities (all P<0.01), and mRNA and protein expressions of HMGA2 were also outstandingly reduced (P<0.01). Conclusion: miR-204 is lowly expressed in RB cell lines; in addition, miR-204 over-expression can suppress RB cell proliferation, migration and invasion, the mechanism of which might be related to down-regulation of the expression of HMGA2.