Coronavirus infection can cause serious respiratory and digestive system diseases in hu-mans and animals.In recent years,the frequent outbreaks and newly outbreaks of coronavirus in-fection have threatened global public health and the development of livestock and poultry.Howev-er,the development of anti-coronavirus drugs and vaccines was restricted due to the insufficient understanding of the mechanisms of coronavirus pathogenesis and cross species transmission.Re-verse genetic manipulation technology is a powerful tool in virological research,which can be used for the study of pathogenesis mechanisms,replication mechanisms and function analysis of protein.It is also be used in the development of attenuated or gene labeled vaccines,and antiviral drugs.Due to the large genome and complex structure of the coronavirus,the reverse genetic manipulation technology of coronavirus has been lagging for a long time.With the continuous updating of molec-ular biology methods,various new construction strategies have emerged.This article focuses on the construction strategy of the reverse genetic operating system for coronavirus,as well as its applica-tion in virus transmission and pathogenic mechanisms,and development of vaccines,which will provide favorable tools for the prevention and control of the coronavirus infection.

Objective:To provide evidence-based recommendations for the prevention and management of lower limb ischemia in veno-arterial extracorporeal membrane oxygenation (VA-ECMO) patients during treatment according to search, evaluate, and summarize the best evidence on the prevention and management of lower limb ischemia in patients with VA-ECMO.Methods:Based on the PIPOST framework (population, intervention, professional, outcome, setting, and type of evidence), an evidence-based question was formulated. A systematic search was conducted according to the "6S" evidence pyramid model in both domestic and international databases, as well as professional association websites, for all evidence related to the prevention and management of lower limb ischemia in VA-ECMO patients (aged ≥18 years). The types of evidence included clinical decisions, guidelines, expert consensus, systematic reviews, evidence summaries, and original studies. The search was conducted from the construction of the databases to February 2024. Two researchers independently conducted a literature quality evaluation, extracted and summarized evidence from the studies that met the quality criteria.Results:A total of 13 articles were included, consisting of 3 clinical decisions, 3 guidelines, 3 expert consensus, 3 systematic reviews, and 1 randomized controlled trial. A total of 18 pieces of evidence in 7 dimensions were summarized, including risk factors of VA-ECMO lower limb ischemia, evaluation before catheterization, evaluation and monitoring during treatment, prevention of lower limb ischemia, treatment of lower limb ischemia, management of distal perfusion catheter (DPC), and monitoring after VA-ECMO weaning.Conclusion:This evidence summary provides evidence-based recommendations for the prevention and management of lower limb ischemia in VA-ECMO patients, aiming to assist clinical healthcare professionals in developing tailored strategies for the prevention and management of lower limb ischemia based on during VA-ECMO support.

Objective: To compare the long-term clinical and radiographic outcomes of transforaminal endoscopic lumbar discectomy (TELD) versus microdiscectomy (MD). Methods: The data of 154 patients with lumbar disc herniation (LDH) who underwent TELD (n = 89) or MD (n = 65) were retrospectively analyzed. The patients’ clinical outcomes were evaluated using visual analogue scales for leg and low back pain, the Japanese Orthopaedic Association (JOA) score, and the Oswestry Disability Index (ODI). The evolution of radiographic manifestations was observed during follow-up. Potential risk factors for a poor clinical outcome were investigated. Results: During a mean follow-up of 5.5 years (range, 5–7 years), the recurrence rate was 4.49% in the TELD group and 1.54% in the MD group. All scores significantly improved from preoperatively to postoperatively in both groups (p < 0.01). The improvement in the ODI and JOA scores was significantly greater in the TELD than MD group (p < 0.05). Forty-seven patients (52.8%) in the TELD group and 32 (49.2%) in the MD group had Modic changes before surgery, most of which showed no changes at the last follow-up. The degeneration grades of 292 discs (71.0%) were unchanged at the last follow-up, while 86 (20.9%) showed improvement, mostly at the upper adjacent segment. No significant difference was observed in the intervertebral height index or paraspinal muscle-disc ratio. Conclusion Both TELD and MD provide generally satisfactory long-term clinical outcomes for patients with LDH. TELD can be used as a reliable alternative to MD with less surgical trauma. Modic type II changes, decreased preoperative intervertebral height, and a high body mass index are predictors of a poor prognosis.

Sonodynamic therapy (SDT) provides a non-invasive therapeutic strategy for killing tumor cells by using low-intensity ultrasound combined with sonosensitizer. Compared with other tumors, urinary system tumors have a good sound window for treatment. Combined with the advantages of low-frequency ultrasound, such as strong penetration and high targeting, SDT has efficient treatment efficiency and broad application prospects in urological tumors.

Objective:To evaluate the role of autophagy in hydrogen-rich solution-induced reduction of remifentanil-induced hyperalgesia in rats.Methods:Thirty-two clean-grade healthy male Sprague-Dawley rats, aged 2-3 months, weighing 240-260 g, were divided into 4 groups ( n=8 each) by a random number table method: incisional pain group (group I), remifentanil+ incisional pain group (group RI), hydrogen-rich solution+ remifentanil+ incisional pain group (group HRI), and hydrogen-rich solution + autophagy inhibitor+ remifentanil+ incisional pain group (MHRI group). The tail vein was catheterized, the equal volume of normal saline was intravenously infused for 60 min while the incisional pain model was developed in group I, and remifentanil was intravenously infused at a rate of 1 μg·kg -1·min -1 for 60 min while the incisional pain model was developed in RI, HRI and MHRI groups, hydrogen-rich solution 10 ml/kg was intraperitoneally injected at 10 min before preparing the model in group HRI, and 3-MA 15 mg/kg was intraperitoneally injected at 1 h before preparing the model in MHRI group, and the other treatments were similar to those previously described in group HRI. The mechanical paw withdrawal threshold (MWT) and thermal paw withdrawal latency (TWL) were determined at 24 h before and 2, 6, 24 and 48 h after the end of infusion. The rats were sacrificed under anesthesia after the behavioral testing, and the lumbar enlargement segment of the spinal cord was removed for determination of the expression of microtubule-associated protein 1 light chain 3Ⅱ (LC3Ⅱ), Beclin-1 and P62 by Western blot. Results:Compared with the baseline at T 0, the MWT was significantly decreased and TWL was shortened at T 1-4 in the four groups ( P<0.05). Compared with group I, the MWT was significantly decreased and TWL was shortened at T 1-4, the expression of LC3 II and Beclin-1 was up-regulated, and the expression of P62 was down-regulated in group RI and group HRI ( P<0.05). Compared with group RI, the MWT was significantly increased and TWL was prolonged at T 1-4 in group HRI and group MHRI, the expression of LC3 II and Beclin-1 was significantly up-regulated, and the expression of P62 was down-regulated in group HRI, and the expression of LC3 II and Beclin-1 was significantly down-regulated, and the expression of P62 was up-regulated in group MHRI ( P<0.05). Compared with group HRI, the MWT was significantly decreased and TWL was shortened at T 1-4, the expression of LC3 II and Beclin-1 was down-regulated, and the expression of P62 was up-regulated in group MHRI ( P<0.05). Conclusions:The mechanism by which hydrogen-rich solution alleviates hyperalgesia may be related to enhancing the level of autophagy in the spinal cord of rats with incisional pain induced by remifentanil.

Objective: To compare the long-term clinical and radiographic outcomes of transforaminal endoscopic lumbar discectomy (TELD) versus microdiscectomy (MD). Methods: The data of 154 patients with lumbar disc herniation (LDH) who underwent TELD (n = 89) or MD (n = 65) were retrospectively analyzed. The patients’ clinical outcomes were evaluated using visual analogue scales for leg and low back pain, the Japanese Orthopaedic Association (JOA) score, and the Oswestry Disability Index (ODI). The evolution of radiographic manifestations was observed during follow-up. Potential risk factors for a poor clinical outcome were investigated. Results: During a mean follow-up of 5.5 years (range, 5–7 years), the recurrence rate was 4.49% in the TELD group and 1.54% in the MD group. All scores significantly improved from preoperatively to postoperatively in both groups (p < 0.01). The improvement in the ODI and JOA scores was significantly greater in the TELD than MD group (p < 0.05). Forty-seven patients (52.8%) in the TELD group and 32 (49.2%) in the MD group had Modic changes before surgery, most of which showed no changes at the last follow-up. The degeneration grades of 292 discs (71.0%) were unchanged at the last follow-up, while 86 (20.9%) showed improvement, mostly at the upper adjacent segment. No significant difference was observed in the intervertebral height index or paraspinal muscle-disc ratio. Conclusion Both TELD and MD provide generally satisfactory long-term clinical outcomes for patients with LDH. TELD can be used as a reliable alternative to MD with less surgical trauma. Modic type II changes, decreased preoperative intervertebral height, and a high body mass index are predictors of a poor prognosis.

Objective: To compare the long-term clinical and radiographic outcomes of transforaminal endoscopic lumbar discectomy (TELD) versus microdiscectomy (MD). Methods: The data of 154 patients with lumbar disc herniation (LDH) who underwent TELD (n = 89) or MD (n = 65) were retrospectively analyzed. The patients’ clinical outcomes were evaluated using visual analogue scales for leg and low back pain, the Japanese Orthopaedic Association (JOA) score, and the Oswestry Disability Index (ODI). The evolution of radiographic manifestations was observed during follow-up. Potential risk factors for a poor clinical outcome were investigated. Results: During a mean follow-up of 5.5 years (range, 5–7 years), the recurrence rate was 4.49% in the TELD group and 1.54% in the MD group. All scores significantly improved from preoperatively to postoperatively in both groups (p < 0.01). The improvement in the ODI and JOA scores was significantly greater in the TELD than MD group (p < 0.05). Forty-seven patients (52.8%) in the TELD group and 32 (49.2%) in the MD group had Modic changes before surgery, most of which showed no changes at the last follow-up. The degeneration grades of 292 discs (71.0%) were unchanged at the last follow-up, while 86 (20.9%) showed improvement, mostly at the upper adjacent segment. No significant difference was observed in the intervertebral height index or paraspinal muscle-disc ratio. Conclusion Both TELD and MD provide generally satisfactory long-term clinical outcomes for patients with LDH. TELD can be used as a reliable alternative to MD with less surgical trauma. Modic type II changes, decreased preoperative intervertebral height, and a high body mass index are predictors of a poor prognosis.

Objective To evaluate the role of autophagy in the treatment of neuropathic pain(NP)with hydrogen-rich saline.Methods Forty adult male Sprague-Dawley rats with successful intubation were randomly divided into 5 groups(n= 8)using a random number table:the sham operation group(group S),the neuropathic pain group(group C),the hydrogen-rich saline group(group H),the autophagy inhibitor group(group M)and the hydrogen-rich saline + autophagy inhibitor group(group HM).There were 8 rats in each group.The NP model was established by chronic constriction of the sciatic nerve(CCI)in rats.The autophagy inhibitor 3-methyladenine(3-MA)was intraperitoneally injected with 30μg/kg in the group M and the group HM.The hydrogen-rich saline(0.6 mmol/L)was intraperitoneally injected with 10 mL/kg in the group H and the group HM.The other groups were intraperitoneally injected with the same amount of normal saline twice a day for 7 consecutive days.Paw withdrawal threshold to mechanical stimulation(MWT)and paw withdrawal latency to thermal stimulation(TWL)were measured at 1 day before and 1,3,5,7 and 14 days after modeling(T0-T5).After the last measurement of pain threshold,the L4-L6 segment of spinal cord was removed for determination of the expression of autophagy-related proteins microtubule-associated protein light chain 3(LC3)Ⅱ,Beclin-1 and p62 proteins by Western blot assay.The expression levels of superoxide dismutase(SOD)and malondialdehyde(MDA)in spinal cord tissue were detected.Results Compared with the group S,MWT and TWL were decreased in the group C at T2-5,the expression levels of LC3 Ⅱ,Beclin-1 and p62 were increased,SOD activity was decreased,and MDA content was increased at T5(P＜0.05).Compared with the group C,MWT and TWL were increased in the group H at T2-5,LC3 Ⅱ and Beclin-1 protein expression levels were increased,p62 protein expression levels were decreased,SOD activity was increased,and MDA content was decreased at T5(P＜0.05).MWT and TWL were decreased in the group M at T2-5,LC3 Ⅱ and Beclin-1 protein expression levels were decreased,p62 protein expression levels were increased,SOD activity was decreased,and MDA content was increased at T5(P＜0.05).Compared with the group M,MWT and TWL were increased in the group HM at T2-5,LC3 Ⅱ and Beclin-1 protein expression levels were increased,p62 protein expression levels were decreased,SOD activity was increased,and MDA content was decreased at T5(P＜0.05).Conclusion Hydrogen-rich saline can alleviate neuropathic pain and inhibit oxidative stress in spinal cord in rats,and the mechanism may be related to the increase of autophagy.

【Objective】 To analyze the application value of penile vibrating perception threshold measurement in the diagnosis of erectile dysfunction (ED) and provide reference for the seversity of ED patients. 【Methods】 The clinical data, Erectile Hardness Scale (EHS) score, International Index of Erectile Function Questionnaire-5 (IIEF-5) score, and the vibrating perception threshold (VPT) of the penis of 257 patients with decreased erectile function as the main complaint or accompanying symptoms treated during Jan. and Dec.2021 were retrospectively collected and analyzed.Patients with EHS=4 and IIEF-5≥22 were classified into the normal group, and the rest into the ED group.The differences in VPT in patients with different EHS scores were compared, and the correlation between IIEF-5 and VPT was analyzed.The diagnostic value of VPT for ED was evaluated with receiver operating characteristic (ROC) curve. 【Results】 The difference in penile VPT among patients with different EHS scores was statistically significant (P<0.05).With the decrease of EHS score, VPT showed an increasing trend.Glans VPT was negatively correlated with IIEF-5 score (ρ=-0.22, P<0.001), and penile shaft VPT was also negatively correlated with IIEF-5 score (ρ=-0.26, P<0.001).The VPT of glans penis [(4.17±1.37) V vs.(3.47±1.24) V, P=0.009] and the VPT of penis body [(3.73±1.41) V vs.(2.99±1.14)V, P=0.003] in the ED group were both higher than those in the normal group.The area under the ROC curve (AUC) of the glans VPT was 0.642.When the cut-off value was 3.537 V, the sensitivity was 63.4%, and the specificity was 63.6%.The AUC of the penile shaft VPT was 0.659.When the cut-off value was 2.775 V, the sensitivity was 72.3%, with a specificity of 54.5%. 【Conclusion】 The penile VPT of ED patients is higher than that of normal ones, and there is a correlation between VPT and the severity of ED.Severe ED is associated with higher VPT.The measurement of penile VPT is helpful for the clinical diagnosis of ED patients.

Abstract Cardiovascular diseases (CVD) have imposed a substantial disease burden in China. Recently, the significance of childhood in CVD prevention has gained a significant attention. Prospective cohort studies have provided conclusive evidence that various risk factors during childhood are associated with the risk of CVD in adulthood. Furthermore, randomized controlled trials have made noteworthy advancements, demonstrating that healthy lifestyles during childhood could significantly promote cardiovascular health. The paper reviews the prevalence characteristics of CVD risk factors in Chinese children, the association with the risk of CVD development in adulthood and the progress of intervention research in childhood, in order to provide a scientific reference for the prevention of CVD from childhood.