Objective To explore the effect and possible mechanism of berberine on the macro-phage polarization of human myeloid leukemia monocytic cell line THP-1 induced by oxidized low-density lipoprotein(ox-LDL).Methods THP-1 cells were induced into macrophages by PMA,and then according to different concentrations of berberine,the cells were divided into con-trol group,and 5,10,20,40 and 50 μmol/L berberine groups.After intervention for 24 or 48 h,CCK8 assay was used to detect cell viability for optimal concentration and time of berberine treat-ment.PMA-induced THP-1 macrophages were assigned into blank group,model group(ox-LDL),berberine group,inhibitor group(phosphatidyl inositol 3-kinase(PI3K)inhibitor LY294002)and berberine+inhibitor group(berberine+LY294002).The contents of inducible nitric oxide syn-thase(iNOS)and TGF-β1 were detected by ELISA.qPCR was employed to measure the mRNA expression of TNF-α,arginase 1(Arg1),PI3K and protein kinase B Akt1,and Western blotting was applied to detect the protein levels of Akt1 and phosphorylated protein kinase B antibody(p-Akt1).Results In 24 h after intervention,the macrophage activity was significantly lower in the 40 and 50 μmol/L berberine groups than the control group(P＜0.05),and after 48 h,the ac-tivity in all the 5 doses of berberine groups was obviously lower than that in the control group[(0.89±0.02)％,(0.82±0.03)％,(0.71±0.02)％,(0.62±0.03)％and(0.53±0.02)％vs(1.01±0.01)％,P＜0.05].Berberine treatment of 20 μmol/L for 24 h had little effect on cell viability,and the dose and the time were regarded as the best concentration and time.Compared with the blank group,iNOS content and TNF-α mRNA level were increased in the model group,while TGF-β1 content,mRNA levels of Arg1,PI3K and Akt1,and p-Akt1/Akt1 protein levels were de-creased(P＜0.05).iNOS content and TNF-α mRNA level were decreased,while TGF-β1 content,mRNA levels of Arg1,PI3K and Akt1 and protein levels of p-Akt1/Akt1s were increased in the berberine group than the model group(P＜0.05).Compared with the berberine group,iNOS con-tent and TNF-α mRNA level were increased,while mRNA levels of Arg1,PI3K and Akt1 and protein levels of p-Akt1/Akt1 were decreased in the berberine+inhibitor group(P＜0.05).Con-clusion Berberine can inhibit the inflammatory response of THP-1 macrophages induced by ox-LDL by activating PI3K/Akt1 pathway,and inhibit the M1 polarization and promote the M2 polarization of macrophages.

Polygala tenuifolia,commonly known as Yuanzhi(YZ)in Chinese,has been shown to possess anti-insomnia properties.However,the material basis and the mechanism underlying its sedative-hypnotic effects remain unclear.Herein,we investigated the active components and neurochemical mechanism of YZ extracts using liquid chromatography tandem mass spectrometry(LC-MS/MS)-based pharmaco-metabolomics and mass spectrometry imaging(MSI)-based spatial resolved metabolomics.According to the results,17 prototypes out of 101 ingredients in the YZ extract were detected in both the plasma and brain,which might be the major components contributing to the sedative-hypnotic effects.Network pharmacology analysis revealed that these prototypes may exert their effects through neuroactive ligand-receptor interaction,serotonergic synapse,dopaminergic synapse,and dopaminergic synapse,among other pathways.LC-MS/MS-based targeted metabolomics and Western blot(WB)revealed that tryptophan-serotonin-melatonin(Trp-5-HT-Mel)and tyrosine-norepinephrine-adrenaline(Tyr-Ne-Ad)are the key regulated pathways.Dopa decarboxylase(DDC)upregulation and phenylethanolamine N-methyltransferase(PNMT)downregulation further confirmed these pathways.Furthermore,MSI-based spatially resolved metabolomics revealed notable alterations in 5-HT in the pineal gland(PG),and Ad in the brainstem,including the middle brain(MB),pons(PN),and hypothalamus(HY).In summary,this study illustrates the efficacy of an integrated multidimensional metabolomics approach in unraveling the sedative-hypnotic effects and neurochemical mechanisms of a Chinese herbal medicine,YZ.

Retrograde adeno-associated viruses (AAVs) are capable of infecting the axons of projection neurons and serve as a powerful tool for the anatomical and functional characterization of neural networks. However, few retrograde AAV capsids have been shown to offer access to cortical projection neurons across different species and enable the manipulation of neural function in non-human primates (NHPs). Here, we report the development of a novel retrograde AAV capsid, AAV-DJ8R, which efficiently labeled cortical projection neurons after local administration into the striatum of mice and macaques. In addition, intrastriatally injected AAV-DJ8R mediated opsin expression in the mouse motor cortex and induced robust behavioral alterations. Moreover, AAV-DJ8R markedly increased motor cortical neuron firing upon optogenetic light stimulation after viral delivery into the macaque putamen. These data demonstrate the usefulness of AAV-DJ8R as an efficient retrograde tracer for cortical projection neurons in rodents and NHPs and indicate its suitability for use in conducting functional interrogations.
Animals ; Haplorhini ; Axons ; Motor Neurons ; Interneurons ; Macaca ; Dependovirus/genetics* ; Genetic Vectors

Here, we report a previously unrecognized syndromic neurodevelopmental disorder associated with biallelic loss-of-function variants in the RBM42 gene. The patient is a 2-year-old female with severe central nervous system (CNS) abnormalities, hypotonia, hearing loss, congenital heart defects, and dysmorphic facial features. Familial whole-exome sequencing (WES) reveals that the patient has two compound heterozygous variants, c.304C>T (p.R102*) and c.1312G>A (p.A438T), in the RBM42 gene which encodes an integral component of splicing complex in the RNA-binding motif protein family. The p.A438T variant is in the RRM domain which impairs RBM42 protein stability in vivo. Additionally, p.A438T disrupts the interaction of RBM42 with hnRNP K, which is the causative gene for Au-Kline syndrome with overlapping disease characteristics seen in the index patient. The human R102* or A438T mutant protein failed to fully rescue the growth defects of RBM42 ortholog knockout ΔFgRbp1 in Fusarium while it was rescued by the wild-type (WT) human RBM42. A mouse model carrying Rbm42 compound heterozygous variants, c.280C>T (p.Q94*) and c.1306_1308delinsACA (p.A436T), demonstrated gross fetal developmental defects and most of the double mutant animals died by E13.5. RNA-seq data confirmed that Rbm42 was involved in neurological and myocardial functions with an essential role in alternative splicing (AS). Overall, we present clinical, genetic, and functional data to demonstrate that defects in RBM42 constitute the underlying etiology of a new neurodevelopmental disease which links the dysregulation of global AS to abnormal embryonic development.
Female ; Animals ; Mice ; Humans ; Child, Preschool ; Intellectual Disability/genetics* ; Heart Defects, Congenital/genetics* ; Facies ; Cleft Palate ; Muscle Hypotonia

By consulting the ancient and moderm literature， this paper makes a textual research on the name， origin， quality evaluation， harvesting and processing of Olibanum， so as to provide a basis for the development of the famous classical formulas containing this medicinal material. According to the herbal textual research， the results showed that Olibanum was first described as a medicinal material by the name of Xunluxiang in Mingyi Bielu（《名医别录》）， until Ruxiang had been used as the correct name since Bencao Shiyi（《本草拾遗》） in Tang dynasty. The main origin was Boswellia carterii from Burseraceae family. The mainly producing areas in ancient description were ancient India and Arabia， while the modern producing areas are Somalia， Ethiopia and the southern Arabian Peninsula. The medicinal part of Olibanum in ancient and modern times is the resin exuded from the bark， which has been mainly harvested in spring and summer. It is concluded that the better Olibanum has light yellow， granular， translucent， no impurities such as sand and bark， sticky powder and aromatic smell. There were many processing methods in ancient times， including cleansing（water flying， removing impurities）， grinding（wine grinding， rush grinding）， frying（stir-frying， rush frying， wine frying）， degreasing， vinegar processing， decoction. In modern times， the main processing methods are simplified to cleansing， stir-frying and vinegar processing. Nowadays， the commonly used specifications include raw， fried and vinegar-processed products. Among the three specifications， raw products is the Olibanum after cleansing， fried products is a kind of Olibanum processed by frying method， vinegar-processed products is the processed products of pure frankincense mixed with vinegar. Based on the research results， it is recommended to select the resin exuded from the bark of B. carterii for the famous classical formulas such as Juanbitang containing Olibanum， processing method should be carried out in accordance with the processing requirements of the formulas， otherwise used the raw products if the formulas without clear processing requirements.

Objective To investigate the characteristics of human papillomavirus (HPV) prevalence and genotype distribution of 18 535 cases in Yuncheng . Methods A sample of 18535 residents who underwent HPV testing in our hospital from August 2020 to September 2023 were enrolled, and HPV genotyping was done to all samples. Then the rate of HPV infection, age distribution, genotype distribution, and multiple infections were statistically analyzed. Results Of the 18,535 subjects included, a total of 4,639 tested positive for HPV, demonstrating a positive rate of 25.03%. The positive rate of HPV infection varied among different age groups (χ²=29.587, P<0.05), with higher rates found in <25 years old group (29.61%) and >60 years old group (25.89%). Overall, 23 genotypes, covering 5315 viruses, were detected. There were 5 low-risk genotypes with the highest percentage of HPV42 (9.29%), and there were 18 high-risk genotypes with HPV52, HPV58, HPV66 and HPV53, subtypes as the most frequent subtypes, accounting for 13.64%, 8.97%, 7.41% and 7.04%, respectively. The type of HPV infection was predominantly single infections, with an overall single infection rate of 21.62% (4008/18535), which accounted for 86.40% (4 008/4 639) of all positive cases, and a multiple genotype infections rate of 3.40% (631/18535). The 25-34 year old group accounted for the largest proportion of single infections (25.12%), while the <25 year old group accounted for the largest proportion of multiple genotype infections (30.74%). Conclusion The prevalence rate of HPV infection in Yuncheng is 25.03%, with a higher positive rate in the <25 years age group and the >60 years age group. A total of 23 HPV genotypes are detected, of which the main genotypes are HPV42, HPV58, HPV66 and HPV53, and the type of infection is dominated by single infections.

[Objective] To evaluate the efficacy and safety of zero-pressure percutaneous nephrolithotomy (PCNL) combined with hands-free pneumatic lithotripsy in the one-stage treatment of calculous pyonephrosis in the non-acute infection phase. [Methods] Clinical data of 15 patients' treated during Feb.2022 and Dec.2023 were retrospectively analyzed.Double-sheath negative pressure PCNL was adopted, with zero-pressure gravity perfusion where the perfusion height was at the level of the patients' kidney, not exceeding 20 cm above the kidney.Pneumatic lithotripsy was performed without the need to hold the device with hand. [Results] The median stone diameter was 28 (range 22—40) mm, with a median stone density of 1028 (range 765—1305) Hu.All operations were successful.The median operation time was 55 (range 35—90) min.Postoperative low fever (Clavien grade Ⅰ) occurred in 1 case, and high fever (Clavien grade Ⅱ) occurred in 1 case, with no complications of Clavien grade Ⅲ or above.The initial stone-free rate was 73.3%, and the stone-free rate after one month was 93.3%. [Conclusion] For patients with calculous pyonephrosis in the non-acute infection phase, one-stage treatment using zero-pressure PCNL combined with hands-free pneumatic lithotripsy demonstrates good efficacy and safety.

[Objective] To summarize the clinical manifestations, pathological characteristics, treatment options and prognosis of the world's first case of prostate ductal adenocarcinoma (PDA) complicated with prostate mucinous adenocarcinoma (PMA). [Methods] The clinical and follow-up data of a patient with PDA and PMA treated in Zhongnan Hospital of Wuhan University were retrospectively analyzed, and relevant literature in PubMed and CNKI databases was retrieved. [Results] The patient sought medical attention due to dysuria, frequent urination, urinary urgency and urinary pain for more than half a year, and was admitted to hospital 3 times in total.The initial diagnosis upon the first admission was benign prostatic hyperplasia complicated with prostatic abscess.After 2 months, the patient was readmitted due to worsening symptoms, received transurethral bladder neck incision+ cystoscopy+ transurethral plasma resection of the prostate, and postoperative diagnosis confirmed PDA with local PMA.Three months after surgery, the patient had bleeding.After auxiliary examinations revealed extensive metastasis, he received hormonal therapy.After 9 months, the patient died due to multiple lung metastases. [Conclusion] Early diagnosis has a significant impact on the treatment and prognosis, but there have been no previous reports of PDA combined with PMA, so the lack of specific biomarkers in the early stage has led to missed diagnosis or misdiagnoses.There is no specific treatment for PDA with PMA. Radical prostatectomy was not satisfactory in the treatment of this case.

Objective:To compare the recovery effect of continuous infusion of dexmedetomidine combined with oxycodone or sufentanil in the anesthesia intensive care unit (AICU) in elderly patients after thoracoscopic radical surgery for lung cancer.Methods:Using the method of prospective study, 80 elderly lung cancer patients underwent selective thoracoscopic radical surgery under general anesthesia in Nanjing First Hospital from February 2021 to May 2022 were selected. The patients were divided into dexmedetomidine combined with sufentanil group (S group) and dexmedetomidine combined with oxycodone group (Q group) by random digits table method with 40 cases each group. On the basis of routine monitoring and treatment after operation, the patients in Q group were continuously injected with oxycodone 0.03 mg/(kg·h) and dexmedetomidine 0.4 μg/(kg·h) through analgesia pump, the patients in S group were continuously injected with sufentanil 0.03 mg/(kg·h) and dexmedetomidine 0.4 μg/(kg·h) through analgesia pump. The wake-up time, extubation time, awakening quality (Aldrete score and bucking score) and comfort level (Bruggrmann comfort scale score, BCS score) after entering the AICU were record; the sedation score (Ramsay score) and pain relief score (numerical rating scale score, NRS score) and hemodynamic changes (mean arterial pressure and heart rate) 3, 5, 7, 10 and 14 h after entering the AICU were record; the level of serum inflammatory factors, including tumor necrosis factor (TNF-α), interleukin-6 (IL-6) and C-reactive protein (CRP) immediately, 5 h and 14 h after entering the AICU; press times of analgesia pump, adverse events, bleeding volume of drainage tube during AICU and overall satisfaction score when leaving the AICU were record.Results:The bucking score in Q group was significantly lower than that in S group: (1.02 ± 0.77) scores vs. (1.88 ± 0.34) scores, the Aldrete score and BCS score were significantly higher than those in S group: (8.93 ± 0.25) scores vs. (5.97 ± 0.32) scores and (3.03 ± 0.32) scores vs. (0.93 ± 0.52) scores, and there were statistical differences ( P<0.01); there were no statistical difference in wake-up time and extubation time between two groups ( P>0.05). There were no statistical difference Ramassy score, NRS score 3 and 5 h after entering the AICU, mean arterial pressure and heart rate between two groups ( P>0.05); the Ramassy score 7, 10 and 14 h after entering the AICU in Q group was significantly lower than that in S group, the NRS score, mean arterial pressure and heart rate were significantly lower than those in S group, and there were statistical differences ( P<0.01). There were no statistical differences in TNF-α, IL-6 and CRP immediately after entering the AICU between two groups ( P>0.05); the TNF-α, IL-6 and CRP 5 and 14 h after entering the AICU in Q group were significantly lower than those in S group, and there were statistical difference ( P<0.01). The press times of analgesia pump, bleeding volume of drainage tube and the incidences of nausea vomiting, respiratory depression, lethargy, restlessness, fever and lung infection in Q group were significantly lower than those in S group: (4.63 ± 1.10) times vs. (18.80 ± 1.54) times, (129.67 ± 4.14) ml vs. (164.00 ± 8.14) ml, 10.0% (4/40) vs. 52.5% (21/40), 2.5% (1/40) vs. 25.0% (10/40), 7.5% (3/40) vs. 47.5% (19/40), 0 vs. 20.0% (8/40), 2.5% (1/40) vs. 22.5% (9/40) and 2.5% (1/40) vs. 20.0% (8/40), and there were statistical differences ( P<0.01 or <0.05); there was no severe hypotension, severe bradycardia and delirium in both groups. The overall satisfaction score in Q group was significantly higher than that in S group: (3.53 ± 0.63) scores vs. (2.70 ± 0.65) scores, and there was statistical difference ( P<0.01). Conclusions:Continuous micro-pump infusion of dexmedetomidine combined with oxycodone in AICU elderly patients with lung cancer after thoracoscopic radical surgery can significantly improve the quality of recovery and comfort during extubation, without affecting the extubation time, and can effectively reduce the degree of pain, stress and inflammatory reaction in the early recovery period, and reduce the incidence of adverse events after surgery.

OBJECTIVE To reveal the role of the basal forebrain(BF)GABAergic neurons in the regulation of isoflurane anesthesia and to elucidate the underlying neural pathways.METHODS The activity of BF GABAer-gic neurons was monitored during isoflurane anesthesia using a genetically encoded calcium indicator in Vgat-Cre mice of both sexes.The activity of BF GABAer-gic neurons was manipulated by chemogenetic and opto-genetic approaches.Sensitivity,induction time and emer-gence time of isoflurane anesthesia were estimated by righting reflex.The electroencephalogram(EEG)power and burst-suppression were monitored by EEG recording.The effects of activation of GABAergic BF-thalamic reticu-lar nucleus(TRN)pathway on isoflurane anesthesia were investigated with optogenetics.RESULTS The activity of BF GABAergic neurons was generally inhibited during isoflurane anesthesia,obviously decreased during the induction of anesthesia and gradually restored during the emergence from anesthesia.Activation of BF GABAergic neurons with chemogenetics and optogenetics promoted behavioral emergence from isoflurane anesthesia,with decreased sensitivity to isoflurane,delayed induction and accelerated emergence from isoflurane anesthesia.Optogenetic activation of BF GABAergic neurons prom-oted cortical activity during isoflurane anesthesia,with decreased EEG delta power and burst suppression ratio during 0.8%and 1.4%isoflurane anesthesia,respectively.Similar to the effects of activating BF GABAergic cell bod-ies,photostimulation of BF GABAergic terminals in the TRN also strongly promoted cortical activation and behav-ioral emergence from isoflurane anesthesia.CONCLU-SION The GABAergic neurons in the BF is a key neural substrate for general anesthesia regulation that facilitates behavioral and cortical emergence from general anesthe-sia via the BF-TRN pathway.