Objective To develop a real-time RT-PCR assay ( rRT-PCR) for efficient detection of duck hepatitis virus type 1 ( DHV-I) .Method According to the different gene sequences of DHV-I from different provinces download from NCBI and to find the conserved sequences.One pair of the specific primers and one TaqMan probe were designed. Then reaction parameters were optimized to develop a real-time RT-PCR assay ( rRT-PCR) .Results This developed rRT-PCR assay could detect 20 template copies of RNA, and its sensitivity was higher than that of the conventional RT-PCR. This rRT-PCR assay was found to be specific and able to detect DHV-I, and no positive results were observed when nucleic acid from Muscovy duck parvovirus, goose parvovims, Newcastle disease and avian influenza virus, egg drop syndrome virus, reticuloendotheliosis virus, duck Tembusu virus, poultry intestinal arc virus were used as rRT-PCR templates.The results of this developed rRT-PCR assay used for 100 duck clinical samples showed a positive rate of 92%, indicating that DHV exists in duck group of Jiangsu province in China.Conclusion This rRT-PCR assay can be used as a rapid tool for detection of DHV-I.

Objective: To study difference of the immune response between oral administration and intraperitoneal injection of total polysaccharide of Sijunzi decoction (SJTP) in mice. Methods: The immunosuppressed mice model was established by intraperitoneal injection of cyclophosphamide (CY). The effects of SJTP on the weight of immune organs,hemolysin response and delayed type hypersensitivity (DTH) were observed,and the influences on normal mouse spleen lymphocytic proliferation of mice were also studied. Results:SJTP can remarkably counteract the immunosuppression induced by CY, decrease the weight loss immune organs,increase the hemolysin response and DTH,and promote the proliferation of lymphocytes.The effect of oral administration is superior to intrapertoneal injection.Conclusion:SJTP can improve the humoral and cellalar immunity of mice.The immune activities resulting from oral administration of SJTP have some special characteristics,and its possible mechanism is also discussed.

BACKGROUND:Prolonged therapy with lamivudine has been associated with tyrosine-methionine-aspartate-aspartate mutation, which results in hepatitis B recurrence. Recently, antiviral agents, such as entecavir, have high efficacy and low resistance rate in hepatitis B-related liver disease. However, the researches on the effect of entecavir in preventing hepatitis B recurrence after liver transplantation are rare. OBJECTIVE:To investigate the effect of entecavir combined with low-dose hepatitis B immunoglobulin in preventing hepatitis B recurrence after liver transplantation. METHODS:The fol ow-up data of 253 patients who had liver transplantation for hepatitis B virus related liver disease were retrospectively analyzed. Al patients received nucleoside analogues therapy formal y before liver transplantation. The effects of entecavir+hepatitis B immunoglobulin and lamivudine+hepatitis B immunoglobulin were compared in al the patients and the patents with hepatitis B recurrence risk factors (positive preoperative HBeAg, DNA-positive hepatitis B virus, hepatoma and tyrosine-methionine-aspartate-aspartate mutation). RESULTS AND CONCLUSION:A total of 253 patients received hepatitis B virus-related liver transplantation, and 29 patients died. There were 202 patients in lamivudine group in which 26 patients were dead and 16 patients had hepatitis B virus recurrence, and the recurrence rate was 7.92%(16/202). However, entecavir group had 51 patients without hepatitis B virus recurrence in which three patients were dead. There were significant differences in the mortality rate and recurrence rate between two groups. Compared with the lamivudine+hepatitis B immunoglobulin, entecavir+hepatitis B immunoglobulin could effectively reduce the recurrence rate of the patients with hepatitis B virus-related risk factors. Hepatitis B immunoglobulin was terminated and nucleoside analogues were modulated when recurrence appeared. Al patients hepatitis B virus DNA were control ed less than 500 IU/mL and liver function returned to normal level. Log-rank test showed that there was no significant difference in the long-term survival rate after timely treatment of hepatitis B virus recurrence. With the prevention of nucleoside analogues combined with hepatitis B immunoglobulin therapy, timely treatment of hepatitis B recurrence has little influence on the prognosis. Entecavir combined with hepatitis B immunoglobulin can effectively prevent the hepatitis B recurrence. For the patients with hepatitis B virus-related risk factors, entecavir combined with hepatitis B immunoglobulin can better reduce the recurrence rate of hepatitis B than lamivudine+hepatitis B immunoglobulin after liver transplantation.

Patients with spinal cord injury is associated with seriously affected gastrointestinal function and imbalance of intestinal flora, leading to increased inflammation of spinal cord nerves. With the proposal of the theory of gut microbiota-gut-brain axis in recent years, the regulatory role of gut microbiota in the central nervous system and gastrointestinal system has gradually attracted attention. Although a considerable number of studies have focused on the effects of intestinal flora characteristics on spinal cord nerve function repair in patients with spinal cord injury from different perspectives, there are numerous research models for treating spinal cord injury with intestinal flora as intervention targets and remains a lack of unified and effective clinical treatment methods. In this paper, the authors review the research progress in characteristics of intestinal microflora and intestinal microflora-targeted therapeutic methods in patients with spinal cord injury, hoping to provide a reference for the clinical treatment and basic research of spinal cord injury.

Objective:To explore the mediating effects of parental perceptions of child vulnerability (PPCV) and child anxiety in the relationship between parental social support and the quality of life in children with epilepsy.Methods:This was a cross-sectional study. Totally 315 children with epilepsy and their 315 parents who visited the neurologic outpatient clinic for video EEG monitoring from September 2021 to April 2022 were selected by convenience sampling and investigated with General Information Questionnaire, Child Vulnerability Scale (CVS), Social Support Rating Scale (SSRS), PROMIS Parents Report Anxiety Short Form, and the Quality of Life in Children with Epilepsy-16 (QOLCE-16). Pearson correlation was used to analyze the relationships between quality of life in children with epilepsy and variables such as parental social support, PPCV, and child anxiety. AMOS 22.0 was utilized to establish a structural equation model to examine the relationships among the variables. A total of 315 questionnaires were distributed, with 295 valid questionnaires retrieved, yielding a response rate of 93.7%.Results:The scores for the 295 parents on the SSRS, CVS, PROMIS Parents Report Anxiety Short Form, and QOLCE-16 were (40.75±10.24), (8.08±4.84), (16.40±5.98), and (72.01±15.43), respectively. Parental social support was negatively correlated with PPCV and child anxiety ( P<0.01), and both PPCV and child anxiety were negatively correlated with the child's quality of life ( P<0.01), while parental social support had a positive correlation with the child's quality of life ( P<0.01). PPCV and child anxiety mediated the relationship between parental social support and the child's quality of life ( P<0.01), with the direct and indirect effects being 0.301 and 0.205 respectively. Conclusions:Parental social support can directly affect the quality of life of children with epilepsy, and also indirectly influence it through PPCV and child anxiety. Clinical nursing staff should provide more care and support for children with epilepsy and their parents, helping to deepen their understanding of the disease, reduce levels of PPCV and child anxiety, thereby enhancing the quality of life for children with epilepsy.

ObjectiveTo investigate the risk factors for tumor recurrence and death after liver transplantation in patients with hepatocellular carcinoma (HCC) and their survival. MethodsThe patients with HCC who underwent liver transplantation in The Fifth Medical Center of Chinese PLA General Hospital from January 2005 to February 2019 were enrolled, and according to the presence or absence of HCC recurrence after liver transplantation, they were divided into recurrence group and non-recurrence group. The t-test or the Mann-Whitney U test was used for comparison of continuous data between two groups, and the chi-square test was used for comparison of categorical data between two groups. Univariate and multivariate Cox proportional-hazards regression model analyses were used to determine the risk factors for HCC recurrence and death after liver transplantation. The Kaplan-Meier method was used for survival analysis, and the receiver operating characteristic (ROC) curve was used to investigate the predictive value of death-related risk factors after liver transplantation. ResultsA total of 391 HCC patients who underwent liver transplantation were enrolled, with a median follow-up time of 2 years, among whom 78(19.95%) experienced HCC recurrence. Preoperative alpha-fetoprotein (AFP) level＞200 ng/ml (recurrence: hazard ratio ［HR］=252, 95% confidence interval ［CI］: 1.58-4.03, P＜0.001; death: HR=2.99, 95%CI: 1.59-5.62, P＜0.001］, total tumor diameter (recurrence: HR=1.20, 95%CI: 1.12-1.28, P＜0.001; death: HR=1.10, 95%CI: 1.02-1.17, P=0.002), and vascular invasion (recurrence: HR=1.15, 95%CI: 1.04-1.26, P=0.016; death: HR=1.10, 95%CI: 1.03-1.18, P=0.004) were independent risk factors for tumor recurrence and death after liver transplantation. The 1-, 5-, and 10-year overall survival rates after liver transplantation were 94.8%, 84.2%, and 83.5%, respectively, and the 1-, 5-, and 10-year disease-free survival rates were 840%, 75.1%, and 75.1%, respectively. AFP, involvement of major blood vessels, body mass index, and total tumor diameter had a certain value in predicting the death of HCC patients with recurrence, with an area under the ROC curve of 0.789 (95% CI: 0.719-0858). ConclusionTumor biological features before transplantation are the key factors for tumor recurrence after transplantation.

Acupuncture can improve the motor and non-motor symptoms of Parkinson's disease, and the effect of acupuncture combined with drug therapy is better than that of drug therapy alone. The possible mechanism includes inhibiting α-synuclein aggregation, oxidative stress, and neuroinflammation, inhibiting the apoptosis of dopaminergic neurons, and achieving a neuroprotective effect. The points mainly selected for Acupuncture treatment for this disease are Zusanli (ST 36), Yanglingquan (GB 34), Taichong (LR 3), Xuehai (SP 10), and other points. Early use of acupuncture and acupuncture combined with medical treatment strategy is worthy of clinical application.

Lymphatic metastasis is commonly encountered in prostate cancer. However, prostate cancer along with lymph node involvement in anterior prostatic fat is rarely reported. Here we present one case. Lymphadenectasis was noticed during robot-assisted laparoscopic radical prostatectomy. Histopathologic examinations confirmed lymph node metastasis in anterior prostatic fat. The patient was assigned to adjuvant androgen deprivation therapy. At 16-month follow-up, biochemical recurrence has not occurred. Detection of lymph node metastasis in anterior prostatic fat of prostate cancer patients provides valuable information for adequate postoperative staging and management.

Purpose: In non-metastatic prostate cancer (nmPCa) setting, it is important to early identify the patients at risk of biochemical recurrence (BCR) for immediate postoperative intervention. Our study aimed to evaluate the potential clinical utility of circulating tumor DNA (ctDNA) for predicting disease recurrence. Materials and Methods: This real-world observational study evaluated 161 cases of nmPCa undergoing next-generation sequencing at our institution. A total of 139 ctDNA samples and 31 biopsied tumor tissue underwent genomic profiling. The study endpoint was BCR after radical prostatectomy. Relationships between the ctDNA status and the biochemical progression-free survival (bPFS) were analyzed by log-rank test and multivariate Cox regression. Results: Of 161 enrolled patients, 19 (11.8%) harbored deleterious alterations in NCOR2, followed by BRCA2 (3.7%), ATR (2.5%), and CDK12 (2.5%). Of available pre-operative blood samples (n=139), ctDNA was detectable in 91 (65.5%). Until last follow-up, 56 of 68 patients (85.3%) with detectable ctDNA had achieved BCR, whereas only eight of 39 patients (20.5%) with undetectable ctDNA had achieved BCR. Patients who had undetectable ctDNA experienced significantly longer bPFS compared with those who had detectable ctDNA (not available vs. 8.2 months; hazard ratio, 0.14; p < 0.01). Pre-operative ctDNA status was a significant prognostic factor of disease recurrence. Conclusion Pre-operative ctDNA detection could identify patients at high risk of recurrence and has the potential to inform immediate postoperative interventions, but these approaches remain to be validated in prospective studies. ctDNA studies can provide insights into accurate monitoring and precise treatment rather than simply following routine clinical care.

Objective:To investigate the clinical efficacy of neoadjuvant chemo-hormonal therapy(NCHT)followed by radical prostatectomy(RP) plus extended pelvic lymphadenectomy for very-high-risk locally advanced prostate cancer.Methods:The data of 327 cases of very-high-risk locally advanced prostate cancer treated in Renji Hospital Affiliated to Shanghai Jiaotong University School of Medicine, The Second Hospital of Tianjin Medical University, and The Third Affiliated Hospital of Sun Yat-sen University from December 2014 to July 2019 were retrospectively analyzed. Patients were divided into two groups according to treatment regimens: the RP group (direct RP + extended pelvic lymphadenectomy 4-6 weeks after the biopsy of prostate) and the NCHT group (4-6 cycles of NCHT prior to RP). There were 171 cases in RP group and 156 cases in NCHT group, respectively. In the RP group, the median age was 67 (ranging 44-83)years. The median PSA at diagnosis was 27.24 (ranging 4.55-207.00) ng/ml. Patients’numbers of clinical T 2, T 3a, T 3b, T 4 stage were 13, 85, 57, 16, respectively, and clinical N 1, N 0 stage were 33 and 138, respectively. Patients’numbers of ISUP grade groups of 1, 2, 3, 4, 5 were 5, 35, 41, 51, 39, respectively. In the NCHT group, The median age was 67 years, ranging 46-78 years. The median PSA at diagnosis was 72.09(ranging 4.08-722.95)ng/ml. Patients’ numbers of clinical T 2, T 3a, T 3b, T 4 stage were 11, 47, 58, 40, respectively, and clinical N 1, N 0stage were 76 and 80, respectively. Patients’numbers of ISUP grade groups of 1, 2, 3, 4, 5 were 1, 11, 33, 43, 68, respectively. At baseline, the NCHT group showed higher PSA, higher ISUP grade, and more advanced clinical stage at diagnosis( P<0.05). The PSA, pathological down-staging rate, and positive surgical margin rate as well as the biochemical recurrence free survival(bRFS)were compared between the two groups. Results:After radical prostatectomy, compared with the RP group, the NCHT group had a higher proportion of patients achieving PSA<0.2 ng/ml at 6-week postoperative follow-up ( P<0.001), a higher pathologic tumor stage down-staging rate ( P<0.001), a higher ISUP down-grading rate ( P<0.001), and a lower positive surgical margins rate ( P<0.001). In addition, 10.9% of the NCHT group achieved pT 0 or minimal residual disease in postoperative pathology exams. Eighty-three patients (48.5%) in the RP group and 125 patients (80.1%) in the NCHT group achieved undetectable PSA after surgery and entered further analysis for bRFS, which showed NCHT group had significantly longer bRFS (19.46 months vs. 6.35 months). NCHT significantly reduced the risk for biochemical recurrence in locally advanced prostate cancer patients( HR=0.278, 95% CI 0.198-0.390, P<0.001). Such a reduce in risk for biochemical recurrence was seen in all subgroups( P<0.001). Conclusions:NCHT might improve surgical outcomes as well as bRFS in very-high-risk locally advanced prostate cancer patients.