Objective:To construct recombinant lentivirus and adenovirus which regulate the expression of c-Cbl gene and evaluate their efficacy.Methods:The interference lentivirus and overexpressed adenovirus targeting human c-Cbl gene were constructed by gene recombination technology.Quantitative PCR and western blotting were used to detect the expression changes in c-Cbl gene and its transcription after leukemia cells(HL60,THP1)were infected by virus.Results:Three recombinant interfering lentiviral vectors targeting human c-Cbl genes to successfully constructed and were identified by DNA sequencing,and the titers of the packaged viruses were all greater than 1 x 108 TU/ml.Among them,shRNA-2 lentivirus had the highest interference efficiency,and the expression of c-Cbl gene and CBL protein were decreased about 95％and 60％respectively after leukemia cells were infected with shRNA-2;In addition,the recombinant overexpression adenovirus targeting human c-Cbl gene was packaged successfully with the virus titer greater than 1 x 109 TU/ml.When leukemia cells were infected with adenovirus,the expression of c-Cbl gene and CBL protein were up-regulated about 10 times and 1.5 times respectively.Conclusion:Both recombinant interfering lentivirus and overexpression adenovirus can efficiently infect leukemia cells and affect the expressions of c-Cbl gene and CBL protein.It will lay a preliminary foundation for the subsequent study on the function of c-Cbl gene in tumor cells.

Objective:To explore the differences in dynamic spontaneous brain activity in individuals with subjective cognitive decline (SCD) and its correlation with spatial navigation ability in SCD subjects.Methods:A total of 72 SCD subjects(SCD group) and 67 normal controls (NC group) matched for age, gender and education level were recruited from September 2020 to February 2023 at the Affiliated Drum Tower Hospital, Medical School of Nanjing University. All participants underwent resting-state functional magnetic resonance imaging (rs-fMRI) examinations, spatial navigation tests and cognitive function assessments. The rs-fMRI time series were segmented using a sliding time window method, and statistical analyses were carried out using SPSS 26.0 software to compare the differences in the dynamic amplitude of low frequency fluctuation (dALFF) between the two groups. Correlation analysis was conducted between dALFF values in different brain regions and scale scores and spatial navigation tests.Results:Compared with the NC group, the dALFF variability in the right precuneus(0.119±0.021, 0.130±0.031) and left cuneus(0.143±0.034, 0.156±0.032) in SCD group decreased ( t=-3.41, -3.12, P<0.05, FDR corrected), and the dALFF variability in the right middle occipital gyrus(0.146±0.023, 0.137±0.020) and right angular gyrus(0.148±0.025, 0.139±0.026) increased ( t=4.51, 3.36, both P<0.05, FDR corrected). The temporal variability of dALFF in the right precuneus in SCD group was negatively correlated with egocentric spatial navigation ( r=-0.341, P=0.025), delayed allocentric spatial navigation ( r=-0.286, P=0.035) and memory function ( r=-0.332, P=0.009). The temporal variability of dALFF in the left middle occipital gyrus was positively correlated with language function ( r=0.339, P=0.015) and visuospatial function ( r=0.343, P=0.008) in SCD group. Conclusions:The temporal variability of dALFF in the right precuneus and the left middle occipital gyrus may be the neurobiological basis of cognitive decline and spatial navigation impairment in SCD subjects, and it can be used as a potential imaging marker for early identification of SCD patients.

Objective To dynamically observe the changes in hypoxia-inducible factor 1α(HIF-1α)and Bcl-2/adenovirus E1B19kDa-interacting protein 3(BNIP3)in children with traumatic brain injury(TBI)and evaluate their clinical value in predicting the severity and prognosis of pediatric TBI.Methods A prospective study included 47 children with moderate to severe TBI from January 2021 to July 2023,categorized into moderate(scores 9-12)and severe(scores 3-8)subgroups based on the Glasgow Coma Scale.A control group consisted of 30 children diagnosed and treated for inguinal hernia during the same period,with no underlying diseases.The levels of HIF-1α,BNIP3,autophagy-related protein Beclin-1,and S100B were compared among groups.The predictive value of HIF-1α,BNIP3,Beclin-1,and S100B for the severity and prognosis of TBI was assessed using receiver operating characteristic(ROC)curves.Results Serum levels of HIF-1α,BNIP3,Beclin-1,and S100B in the TBI group were higher than those in the control group(P＜0.05).Among the TBI patients,the severe subgroup had higher levels of HIF-1α,BNIP3,Beclin-1,and S100B than the moderate subgroup(P＜0.05).Correlation analysis showed that the serum levels of HIF-lα,BNIP3,Beclin-1,and S100B were negatively correlated with the Glasgow Coma Scale scores(P＜0.05).After 7 days of treatment,serum levels of HIF-1α,BNIP3,Beclin-1,and S100B in both non-surgical and surgical TBI patients decreased compared to before treatment(P＜0.05).ROC curve analysis indicated that the areas under the curve for predicting severe TBI based on serum levels of HIF-1α,BN1P3,Beclin-1,and S100B were 0.782,0.835,0.872,and 0.880,respectively(P＜0.05),and for predicting poor prognosis of TBI were 0.749,0.775,0.814,and 0.751,respectively(P＜0.05).Conclusions Serum levels of HIF-1α,BNIP3,and Beclin-1 are significantly elevated in children with TBI,and their measurement can aid in the clinical assessment of the severity and prognosis of pediatric TBI.IChinese Journal of Contemporary Pediatrics,2024,26(4):378-384]

Acupuncture, a therapeutic treatment defined as the insertion of needles into the body at specific points (ie, acupoints), has growing in popularity world-wide to treat various diseases effectively, especially acute and chronic pain. In parallel, interest in the physiological mechanisms underlying acupuncture analgesia, particularly the neural mechanisms have been increasing. Over the past decades, our understanding of how the central nervous system and peripheral nervous system process signals induced by acupuncture has developed rapidly by using electrophysiological methods. However, with the development of neuroscience, electrophysiology is being challenged by calcium imaging in view field, neuron population and visualization in vivo. Owing to the outstanding spatial resolution, the novel imaging approaches provide opportunities to enrich our knowledge about the neurophysiological mechanisms of acupuncture analgesia at subcellular, cellular, and circuit levels in combination with new labeling, genetic and circuit tracing techniques. Therefore, this review will introduce the principle and the method of calcium imaging applied to acupuncture research. We will also review the current findings in pain research using calcium imaging from in vitro to in vivo experiments and discuss the potential methodological considerations in studying acupuncture analgesia.
Calcium ; Acupuncture Therapy ; Acupuncture ; Acupuncture Analgesia/methods* ; Acupuncture Points ; Technology

Plant natural products have a wide range of pharmacological properties, not only can they be used as plant dietary supplements to meet the nutritional needs of the human body in the accelerated pace of life, but also occupy an important position in the research and development of therapeutic drugs for the treatment of tumors, inflammation and other diseases, and have been widely accepted by the public due to their good safety. However, despite the above advantages of plant natural products, limiting factors such as low solubility, poor stability, lack of targeting, high toxicity and side effects, and unacceptable odor have greatly impeded their conversion to clinical applications. Therefore, the development of new avenues for the application of new natural products has become an urgent problem to be solved at present. In recent years, with the continuous development of research, various strategies have been developed to improve the bioavailability of natural products. Among them, nanocarrier delivery system is one of the most attractive strategies at present. In past studies, a large number of nanomaterials (organic, inorganic, etc.) have been developed to encapsulate plant-derived natural products for their efficient delivery to specific organs and cells. Up to now, nanotechnology has not only been limited to pharmaceutical applications, but is also competing in the fields of nanofood processing technology and nanoemulsions. Among the various nanocarriers, liposomes are the largest nanocarriers with the largest market share at present. Liposomes are bilayer nanovesicles synthesized from amphiphilic substances, which have advantages such as high drug loading capacity and stability. Attractively, the flexible surface of liposomes can be modified with various functional elements. Functionalized modification of liposomes with different functional elements such as antibodies, nucleic acids, peptides, and stimuli-responsive moieties can bring out the excellent drug delivery function of liposomes to a greater extent. For example, the modification of functional elements with targeting function such as nucleic acids and antibodies on the surface of liposomes can deliver natural products to the target location and improve the bioavailability of drugs; the modification of stimulus-responsive groups such as photosensitizers, magnetic nanoparticles, pH-responsive groups, and temperature sensitizers on the surface of liposomes can achieve controlled release of drugs, localized targeting, and synergistic thermotherapy. In addition to the above properties, by using functionalized liposomes to encapsulate natural products with irritating properties can also effectively mask the irritating properties of natural products, improve public acceptance, and increase the possibility of application of irritating natural products. There are various strategies for modifying liposomes with functional elements, and the properties of functionalized liposomes constructed by different construction strategies differ. The commonly used construction strategies for functionalized liposomes include covalent modification and non-covalent modification. These two types of construction strategies have their own advantages and disadvantages. Covalent modification has better stability than non-covalent modification, but its operation is cumbersome. With the above background, this review focuses on the three typical problems faced by plant natural products at present, and summarizes the specific applications of functionalized liposomes in them. In addition, this paper summarizes the construction strategies for building different types of functionalized liposomes. Finally, this paper will also review the opportunities and challenges faced by functionalized liposomes to enter clinical therapy, and explore the opportunities to overcome these problems, with a view to better realizing the precise control of plant nanomedicines, and providing ideas and inspirations for researchers in related fields as well as relevant industrial staff.

ObjectiveTo investigate the efficacy and safety of N-acetylcysteine （NAC） in the treatment of severe alcoholic hepatitis （SAH）， and to provide a basis for clinical medication for SAH. MethodsA prospective randomized controlled trial was conducted among 172 SAH patients with a Maddrey discriminant function score of >32 points who were recruited by The Fifth Medical Center of Chinese PLA General Hospital from June 2015 to June 2018， and these patients were divided into NAC group with 84 patients and control group with 86 patients. NAC （8 g/day， 28 days） was assessed in terms of its safety in SAH patients， its impact on 28-day biochemical parameters， and its role in improving 28- and 180-day survival rates. A further analysis was performed to investigate the effect of NAC on the 28- and 180-day survival rates of SAH patients with acute-on-chronic liver failure （ACLF-SAH patients） and those without acute-on-chronic liver failure （non-ACLF-SAH patients）. The independent-samples t test was used for comparison of normally distributed continuous data between two groups， and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between two groups. The Kaplan-Meier method was used to plot survival curves， and the Log-rank test was used for comparison of survival curves. Univariate and multivariate Cox proportional-hazards regression model analyses were used to investigate independent influencing factors. ResultsNo serious adverse events were observed during NAC treatment， suggesting that NAC had a good safety profile. Compared with the control group， NAC did not significantly improve the 28-day biochemical parameters （all P>0.05） and survival rate of SAH patients （P=0.081）， but it could improve the 180-day survival rate of SAH patients （67.4% vs 81.0%， χ²=4.280， P=0.039）. NAC did not improve the 28- and 180-day survival rates of ACLF-SAH patients （both P>0.05）； NAC did not improve the 28-day survival rate of non-ACLF-SAH patients （P>0.05）， but it could improve the 180-day survival rate of these patients （68.4% vs 88.9%， χ²=4.883， P=0.027）. The multivariate Cox regression survival analysis showed that NAC treatment （hazard ratio ［HR］=2.530， 95% confidence interval ［CI］： 1.334‍ ‍— 4.796， P=0.004，）， Maddrey discriminant function score （HR=3.852， 95%CI： 2.032 — 7.304， P<0.001）， and serum sodium level （HR=1.948， 95%CI： 1.079‍ ‍—‍ ‍3.517， P=0.027） were independent influencing factors for 180-day survival rate in SAH patients. ConclusionNAC has a good safety profile in the treatment of SAH and can improve the 180-day survival rate of SAH patients， and in particular， non-ACLF-SAH patients can benefit from NAC treatment in terms of middle- and long-term survival rates.

AIM: To investigate the clinical effect and stability of biological amnion membrane coverage combined with corneal bandage lens for patients with chemical burns of ocular surface.METHODS: Retrospective study. The clinical data of 49 cases(49 eyes)of patients with chemical burns of ocular surface treated in our hospital between December 2018 and August 2021 were collected. They were divided into the biological amnion membrane coverage group and the biological amnion membrane coverage combined with corneal bandage lens group according to the surgical method. The loss time of biological amniotic membrane, postoperative pain score, repair rate and time of ocular surface, visual acuity and complications were compared between the two groups.RESULTS: The ocular pain scores of the two groups were 2.208±0.758(the biological amnion membrane coverage group)and 2.063±0.800(the biological amnion membrane coverage combined with corneal bandage lens group)at 1d after surgery, respectively(P>0.05). But at the 3d and 7d after surgery, pain scores were 1.844±0.762 and 1.150±0.582, 1.684±0.820 and 0.750±0.514, respectively(all P<0.05). The loss time of biological amniotic membrane in the biological amnion membrane coverage combined with corneal bandage lens group was 10.75±2.63d, which was longer than that in the biological amnion membrane coverage group(7.60±2.22d; P<0.05). Moreover, it has better ocular surface repair effect than the biological amnion membrane coverage group. At 6mo after operation, visual acuity in the two groups was 0.30(0.10, 0.55)and 0.30(0.20, 0.58), respectively(P>0.05). Additionally, there was no differences in the complications of both groups(P>0.05).CONCLUSION:Biological amnion membrane coverage has effective treatment effects on patients with chemical burns of ocular surface, and it can delay the loss time of biological amniotic membrane, increase repair rate of ocular surface and alleviate postoperative discomfort if combined with corneal bandage lens.

OBJECTIVE: To observe the clinical efficacy of Miao medicinal crossbow acupuncture therapy as adjuvant treatment for lung cancer pain based on oxycodone hydrochloride extended-release tablet. METHODS: A total of 60 patients with lung cancer pain were randomized into an observation group (30 cases, 1 case dropped off) and a control group (30 cases). In the control group, oxycodone hydrochloride extended-release tablet was given orally, 10 mg a time, once every 12 hours. On the basis of the treatment in the control group, Miao medicinal crossbow acupuncture therapy was applied once every other day in the observation group. The treatment of 14 days was required in the two groups. Before and after treatment, the numerical rating scale (NRS) score, number of break-out pain and Karnofsky performance status (KPS) score were observed in the two groups. The equivalent oxycodone consumption and rate of adverse reactions were recorded, the analgesic effect was evaluated in the two groups. RESULTS: Compared before treatment, the NRS scores and number of break-out pain were decreased while the KPS scores were increased after treatment in the two groups (P<0.01). After treatment, the NRS score and number of break-out pain in the observation group were lower than the control group (P<0.01), the KPS score in the observation group was higher than the control group (P<0.05). The equivalent oxycodone consumption of whole course and the rate of adverse reactions i.e. constipation, drowsiness, nausea and vomiting in the observation group were lower than the control group (P<0.05). The analgesic effect rate was 93.1% (27/29) in the observation group, which was superior to 63.3% (19/30) in the control group (P<0.05). CONCLUSION On the basis of oxycodone hydrochloride extended-release tablet, Miao medicinal crossbow acupuncture therapy as adjuvant treatment can effectively relieve the pain degree, reduce the number of break-out pain and improve the health status and quality of life in patients with lung cancer pain, enhance the efficacy of medication and reduce its adverse reactions.
Humans ; Cancer Pain ; Oxycodone ; Quality of Life ; Lung Neoplasms ; Pain ; Acupuncture Therapy ; Adjuvants, Immunologic ; Lung ; Analgesics

Existing traditional Chinese medicine (TCM)-related databases are still insufficient in data standardization, integrity and precision, and need to be updated urgently. Herein, an Encyclopedia of Traditional Chinese Medicine version 2.0 (ETCM v2.0, http://www.tcmip.cn/ETCM2/front/#/) was constructed as the latest curated database hosting 48,442 TCM formulas recorded by ancient Chinese medical books, 9872 Chinese patent drugs, 2079 Chinese medicinal materials and 38,298 ingredients. To facilitate the mechanistic research and new drug discovery, we improved the target identification method based on a two-dimensional ligand similarity search module, which provides the confirmed and/or potential targets of each ingredient, as well as their binding activities. Importantly, five TCM formulas/Chinese patent drugs/herbs/ingredients with the highest Jaccard similarity scores to the submitted drugs are offered in ETCM v2.0, which may be of significance to identify prescriptions/herbs/ingredients with similar clinical efficacy, to summarize the rules of prescription use, and to find alternative drugs for endangered Chinese medicinal materials. Moreover, ETCM v2.0 provides an enhanced JavaScript-based network visualization tool for creating, modifying and exploring multi-scale biological networks. ETCM v2.0 may be a major data warehouse for the quality marker identification of TCMs, the TCM-derived drug discovery and repurposing, and the pharmacological mechanism investigation of TCMs against various human diseases.

Objective: To analyze the association between sleep duration and cognitive function of the elderly in six provinces of China. Methods: Based on the cross-sectional survey data of the elderly from the Healthy Ageing Assessment Cohort Study in 2019, 4 644 participants' sociodemographic and economic indicators, lifestyle, prevalence of major chronic diseases, and sleep status, including night-time sleep duration, daytime sleep duration and insomnia, were collected by questionnaires. Cognitive function was evaluated by the Mini-Mental State Examination. Multivariate logistic regression was used to analyze the association between night-time sleep duration, daytime sleep duration and cognitive function. Results: The mean age of 4 644 respondents was (72.3±5.7) years, and 2 111 of them were males (45.5%). The mean total daily sleep time of the elderly was (7.9±1.9) hours, and the proportion of those who slept less than 7.0, 7.0-8.9 and≥9.0 hours was 24.1% (1 119), 42.1% (1 954) and 33.8% (1 571), respectively. The mean sleep time at night was (6.9±1.7) hours. About 23.7% (1 102) of the elderly did not sleep during the day, and the mean duration of the elderly who slept during the day was (78±51) minutes. Among the elderly with insomnia, 47.9% were still satisfied with their sleep quality. The mean value of MMSE score of 4 644 respondents was (24.5±5.3), and the cognitive impairment rate was 28.3% (1 316). The results of multivariate logistic regression model analysis showed that the OR (95%CI) value of the risk of cognitive impairment in older people who did not sleep, slept for 31 to 60 minutes and slept more than one hour was 1.473 (1.139 to 1.904), 1.277 (1.001 to 1.629) and 1.496 (1.160 to 1.928), respectively, compared with those who slept for 1 to 30 minutes during the daytime. Compared with those who slept for 7.0‒8.9 hours at night, the OR (95%CI) value of the risk of cognitive impairment in older people who slept more than 9.0 hours was 1.239 (1.011 to 1.519). Conclusion: The cognitive function is related to sleep duration in the Chinese elderly.
Male ; Humans ; Aged ; Female ; Sleep Initiation and Maintenance Disorders/complications* ; Cross-Sectional Studies ; Cohort Studies ; Sleep ; Sleep Wake Disorders ; Cognition ; China/epidemiology*