Hepatitis C virus (HCV), one of the major pathogens of viral hepatitis, causes significant hazards in humans. Interferon treatment in combination with ribavirin is used as the first line clinical treatment for HCV infection. However, good response to this treatment has only been observed in few patients and repeated recurrence has also been reported frequently. Therefore, new antiviral agents and therapies are in urgent demand. Here, we report a newly constructed Escherichia coli RNase P based M1GS ribozyme that can specifically and efficiently target the core gene of HCV. The guide sequence (GS) of this M1IGS was designed according to the sequence of the core coding region of HCV genome. The GS was then covalently linked to the 3' terminus of M1 RNA, the catalytic subunit of RNase P from Escherichia coli. The specification of this sequence-specific ribozyme, M1GS, was then examined using an in vitro cleavage assay. The cytotoxicity and its activity in inhibition of HCV gene expression and viral proliferation were further studied in vivo. Our results show that the reconstructed M1GS ribozyme displayed obvious catalytic activity in cleaving target mRNAs fragment in vitro. Notable reduction in the expression of HCV core protein and a 1 000-fold reduction in viral growth were also observed in cultured HCV infected Huh7.5.1 cells expressing the functional M1GS ribozyme. This study demonstrated a direct evidence for the antiviral activity of the customized M1GS-HCV/C141 ribozyme, and thus provided a promising new strategy for clinical treatment of HCV infection.
Antiviral Agents ; pharmacology ; Escherichia coli ; genetics ; Genetic Engineering ; Hepacivirus ; genetics ; physiology ; RNA, Catalytic ; genetics ; pharmacology ; RNA, Guide ; genetics ; Ribonuclease P ; genetics ; Viral Core Proteins ; genetics

Objective:To investigate the improvement effect of acupuncture combined with early rehabilitation training on clinical efficacy, traditional Chinese medicine syndrome score, and neurological and motor functions of patients with acute cerebral infarction.Methods:A total of 76 patients with acute cerebral infarction who received treatment in the Department of Neurology at Jinhua Municipal Central Hospital from June 2019 to June 2022 were included in this prospective study. These patients were randomly divided into an observation group and a control group using the random number table method, with 38 patients in each group. The control group received early rehabilitation training, while the observation group received acupuncture combined with early rehabilitation training. The clinical efficacy was compared between the two groups. Pre- and post-treatment traditional Chinese medicine symptom scores and neurological and motor functions were compared between the two groups.Results:The overall response rate in the observation group was 89.47% (34/38), which was significantly higher than 67.59% (25/38) in the control group ( Z = 2.41, P = 0.016). After treatment, the scores of hemiplegia, mouth and eye deviation, and speech difficulty in both groups were significantly decreased compared with before treatment, and the observation group showed a greater degree of decline in these indices than the control group ( t = 4.73, P < 0.001; t = -10.58, P < 0.001; t = 6.42, P < 0.001). After treatment, the neurological deficit scale scores in each group were decreased compared with before treatment, and the observation group showed a greater degree of decline compared with the control group ( t = -7.33, P < 0.001). After treatment, the Fug-Meyer motor function scale and Barthel index scores in each group were significantly increased compared with before treatment, and the observation group showed a greater degree of increases compared with the control group ( t = 3.72, P < 0.001; t = -5.02, P < 0.001). After treatment, the bare hand muscle strength scores of the core muscles in both groups of patients were increased compared with before treatment, and the observation group showed a greater improvement compared with the control group ( t = 3.31, P < 0.001). Conclusion:Acupuncture combined with rehabilitation training can enhance clinical efficacy, improve traditional Chinese medicine syndrome scores, enhance neurological and motor functions, and strengthen core muscle strength in patients with acute cerebral infarction. This treatment approach is worthy of being further promoted in clinical practice.

Objective To investigate the risk factors of cardiovascular disease among elderly patients in Hangzhou and to assess the level of risk of cardiovascular events in 10 years, so as to provide a reliable basis for preventing, reducing and delaying the incidence of cardiovascular events among elderly patients. Methods From June 2015 to June 2016, we selected 8336 elderly patients with health check-up of medical examination center in two ClassⅢ Grade A hospitals by baseline survey. The Self-Designed Living Behavior Pattern Questionnaire was used to investigate the behavior pattern of living such as smoking. TheGuidelines for Prevention of Cardiovascular Disease by WHO/ISH was used to assess the risk of cardiovascular events in 10 years combined with physical examination data.Results Among 8336 elderly patients with health check-up, there were more than 46.34％ of the elderly patients with incidence of cardiovascular events in 10 years for higher than 40％. There were statistical differences in levels of risk of cardiovascular events among patients with different sexes and ages (P<0.01), and male and the aged were the susceptible population. Among the risk factors of cardiovascular events, the abnormal rate of blood pressure, cholesterol and blood glucose was 72.75％, 67.11％ and 39.71％ respectively.Conclusions The level of risk of cardiovascular events in 10 years among elderly patients in Hangzhou is high. Prevention and intervention should be carried out aiming at susceptible population and risk factors.

Objective:To determine the median effective dose (ED 50) of 0.5% ropivacaine based on femoral nerve cross-sectional area for ultrasound-guided femoral nerve block. Methods:Patients of both sexes, aged 18-64 yr, of American Society of Anesthesiologists physical status I or Ⅱ, with body mass index of 20-30 kg/m 2, scheduled for elective open reduction and internal fixation for patella fracture or removal of patella fracture by internal fixation, were enrolled in this study.Ultrasonic localization of femoral nerve was performed for measurement of the femoral nerve cross-sectional area, and 0.5% ropivacaine was injected based on the area.ED 50 was determined by Dixon′s up-and-down sequential method.The initial dose was 0.22 ml/mm 2, and the difference between the two successive doses was 0.02 ml/mm 2.The effective block was defined as complete loss of pain sensation in the areas of anterior skin of knee joint, skin on the inner side of the calf and dorsal medial skin of the foot and the degree of motor block was in stages 1-3 assessed using Brunnstrom motor function within 30 min after nerve block.Nerve block was considered ineffective if pain occurred in any nerve distribution area mentioned above.The study was terminated if 7 effective and ineffective alternating waves occurred.ED 50 and 95% confidence interval (CI) were calculated using Probit analysis. Results:Twenty-seven patients were enrolled in the study with the femoral nerve cross-sectional area (75±5) mm 2.ED 50 (95%CI) of 0.5% ropivacaine for ultrasound-guided femoral nerve block was 0.106 (0.069-0.125) ml/mm 2. Conclusion:ED 50 of 0.5% ropivacaine based on femoral nerve cross-sectional area for ultrasound-guided femoral nerve block is 0.106 ml/mm 2.

Objective To analyze the current status and influencing factors of demoralization syndrome in patients with liver cirrhosis at the decompensated stage.Methods A total of 444 pa-tients with liver cirrhosis at the decompensated stage were selected as study subjects from four tertiary hospitals in Changsha,Hunan Province using a convenient sampling method.The survey was conduc-ted using a general information questionnaire,the Chinese version of the Demoralization Scale,the Social Support Rating Scale,the Chinese version of the Brief Illness Perception Questionnaire,and the Simplified Coping Style Questionnaire.Results The incidence of demoralization syndrome in pa-tients with liver cirrhosis at the decompensated stage was 71.62％,with a mean total score of(38.21±14.34)for the Demoralization Scale.Multiple linear regression analysis showed that prima-ry disease,the number of complications,Child-Pugh classification,coping style,illness perception,and social support were influencing factors of demoralization syndrome in patients with liver cirrhosis at the decompensated stage(P＜0.05).Conclusion The demoralization syndrome in patients with decompensated cirrhosis is at a severe level,and its main influencing factors are primary disease,the number of complications,Child-Pugh classification,social support,illness perception,and coping style.Healthcare professionals should regularly assess the level of demoralization syndrome in patients with liver cirrhosis at decompensated stage and adopt targeted interventions to improve their psycholog-ical status.

Objective To analyze the current status and influencing factors of demoralization syndrome in patients with liver cirrhosis at the decompensated stage.Methods A total of 444 pa-tients with liver cirrhosis at the decompensated stage were selected as study subjects from four tertiary hospitals in Changsha,Hunan Province using a convenient sampling method.The survey was conduc-ted using a general information questionnaire,the Chinese version of the Demoralization Scale,the Social Support Rating Scale,the Chinese version of the Brief Illness Perception Questionnaire,and the Simplified Coping Style Questionnaire.Results The incidence of demoralization syndrome in pa-tients with liver cirrhosis at the decompensated stage was 71.62％,with a mean total score of(38.21±14.34)for the Demoralization Scale.Multiple linear regression analysis showed that prima-ry disease,the number of complications,Child-Pugh classification,coping style,illness perception,and social support were influencing factors of demoralization syndrome in patients with liver cirrhosis at the decompensated stage(P＜0.05).Conclusion The demoralization syndrome in patients with decompensated cirrhosis is at a severe level,and its main influencing factors are primary disease,the number of complications,Child-Pugh classification,social support,illness perception,and coping style.Healthcare professionals should regularly assess the level of demoralization syndrome in patients with liver cirrhosis at decompensated stage and adopt targeted interventions to improve their psycholog-ical status.