This paper comprehensively discusses on the potential neurobiological mechanisms of acupuncture in the treatment of tobacco dependence， focusing on three important aspects， including acupuncture's regulation of tobacco dependence behavior， effects of acupuncture on withdrawal syndrome， and the role of acupuncture in preventing relapse. It is found that acupuncture can inhibit drug-seeking behavior by regulating the reward pathway and related neurons， such as dopamine， thus modulating tobacco dependence behavior. It also alleviates withdrawal symptoms by improving the oral environment of smokers and reducing negative emotions after quitting. Furthermore， acupuncture can prevent relapse by decreasing brain network activity related to smoking cravings and improving cognitive brain functions like addiction memory. Currently， research on the specific neurobiological mechanism of acupuncture in treating tobacco dependence and the involved neural circuits is limited. Future research directions are proposed， including the evaluation of clinical effects， exploration of specific therapeutic mechanisms， investigation of brain pathology， and strengthening the exploration of brain functions. Additionally， combining modern technologies to clarify the neural circuits involved in acupuncture intervention will provide a basis for acupuncture treatment of tobacco addiction.

Objective:To reveal the molecular characteristics of mononuclear phagocytes (MPs) in rat model of peripheral nerve injury (PNI) using single-cell RNA sequencing (scRNA-seq) technology that would provide the developmental changes and major biological process involved in the function of MPs after PNI.Methods:Twenty-seven male SD rats (200-300 g in weight) were selected from the Department of Hand and Foot Microscopy and Wound Repair Surgery, Henan Provincial People's Hospital (People's Hospital of Zhengzhou University) and the Department of Orthopaedics of First Affiliated Hospital of Zhengzhou University from July 2023 to December 2023. The rats were divided into a Sham operation group (Sham group), a 3 days post crush group (3 dpc group) and a 7 days post crush group (7 dpc group), following the randomised table method with 9 rats per group. After 7 days of environmental acclimatisation, the 3 dpc group and 7 dpc group were subjected to have the right sciatic nerve crushed in order to create a model of crush injury. And as a control group, the Sham group was subjected to Sham surgery only. Nine right sciatic nerves of rats were collected from each group at the corresponding time pints. Single-cell isolation was performed on the 10X Genomics platform. ScRNA-seq libraries were constructed using the Gel Bead Kit V3 and the libraries were sequenced using an Illumina Novaseq 6000 sequencer. Dimensionality reduction was performed using Principal Component Analysis and T-Distributed Stochastic Neighbor Embedding to visualise and explore the cellular heterogeneity within the dataset. Nine distinct cell clusters and their corresponding marker genes were identified based on the dimensionality-reduced data. Differential gene expression analysis was then performed to identify differentially expressed genes (DEGs) in MPs between different groups. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analysis were performed to uncover the biological processes and pathways based on the DEGs. Monocle program for pseudo-time analysis was used to infer the developmental trajectory of MPs after injury.Results:A total of 19 054 cells were obtained by sequencing, and the results showed that the proportion of MPs in peripheral nerves was significantly up-regulated after PNI, and MPs were classified into 9 cellular subgroups based on the clustering analysis of the scRNA-seq data, which were Cluster 1 (3 398 cells), Cluster 2 (3 388 cells), Cluster 3 (3 262 cells), Cluster 4 (2 825 cells), Cluster 5 (2 753 cells), Cluster 6 (1 894 cells), Cluster 7 (648 cells), Cluster 8 (492 cells) and Cluster 9 (394 cells), respectively. Based on the expression of different cell subpopulation markers, MPs in the Sham group, 3 dpc group and 7 dpc group of sciatic nerves were classified into 9 cell clusters and the distributions of different MPs clusters in the 9 sciatic nerve samples were identified, among which, the Sham group had the lowest number of MPs cells in the sciatic nerve samples (a total of 2 719 cells) and the clusters were mainly dominated by clusters 5 (1 119 cells) and clusters 6 (1 240 cells). The 3 dpc group had the highest number of MPs cells (9 760 cells in total) and the clusters were mainly dominated by cluster 2 (1 760 cells), cluster 3 (3 130 cells) and cluster 4 (2 300 cells). The MPs (6 575 cells in total) in the 7 dpc group were mainly dominated by cluster 1 (2 406 cells) and cluster 2 (1 628 cells). Compared with the Sham group, the GO and KEGG annotations of the DEGs were significantly upregulated in the 3 dpc group, indicating that MPs in the rat sciatic nerves would have the ability to bind to extracellular molecules and remove debris from the injury site at 3 days post-injury, and the 7 dpc group would have the ability to activate the signalling pathways related to nerve repair. The proposed time-series analysis revealed that, in the uninjured condition, the MPs were mainly in the cluster 5 (Ccl17 +Cd80 +) and cluster 6 (Fcmr +Slc9a9 +). At 3 days post-injury, MPs developed into cell types dominated by cluster 2 (Cd8b +Meis3 +), cluster 3 (Il10 +Cd163 +) and cluster 4 (Ccl24 +Prg4 +). At 7 days post-injury, the effector state of cluster 2 among the main cell types of MPs was still maintained but the other parts had developed into cluster 1 (Hspa1b +Apobec1 +) related phenotypes. Conclusion:The molecular characteristics of MPs in the peripheral nerve revealed through scRNA-seq data provide valuable insights into the role of MPs in mediating inflammation and neural regeneration after PNI.

Objective:To investigate the role and mechanism of N 6-methyladenosine (m 6A) methyltransferase-like 3 (METTL3) in vascular calcification (VC) of chronic kidney disease (CKD) through apoptosis-associated protein. Methods:(1) Real-time fluorescence quantitative PCR was used to test METTL3 mRNA in serum of maintenance hemodialysis (MHD) patients. (2) Western blotting was used to detect the expression of METTL3 protein in high-phosphorus stimulated vascular smooth muscle cells (VSMCs), and immunofluorescence double lable was used to observe the distribution of METTL3 and Runt-related transcription factor 2 (Runx2). The METTL3 overexpressed and knockdown plasmids were constructed and transfected into VSMCs. Alizarin red staining was used to detect calcification degree. Western blotting was used to detect the expressions of osteogenic markers [Runx2, bone morphogenetic protein-2(BMP-2), collagen Ⅰ] and apoptosis- related proteins Bax and Bcl-2. (3) SD rats were randomly divided into control group, CKD-VC group and S-adenosylhomocysteine (SAH) intervention group. The calcification of thoracic aorta was evaluated by von Kossa staining, and the protein expressions of Runx2, Bax and Bcl-2 were detected by immunohistochemistry and Western blotting.Results:(1) METTL3 mRNA expression in MHD patients with VC was significantly lower than that in non-VC patients ( P<0.05), and was negatively correlated with coronary artery calcium score ( r=-0.65, P<0.001). (2) The expression of METTL3 in VSMCs stimulated by high phosphorus was decreased and showed a time dependence. Immunofluorescence double label showed that METTL3 and Runx2 were co-expressed in the nucleus. METTL3 was overexpressed in high-phosphorus induced VSMCs, and the expressions of Runx2, collagen I and BMP-2 were significantly decreased, accompanied by the decrease of calcified nodules and Bax/Bcl-2 ratio (all P<0.05). Conversely, METTL3 knockdown aggravated VSMCs calcification by inducing apoptosis. (3) Furthermore, METTL3 inhibitor SAH was administered in vivo, and it was found that inhibition of METTL3 expression significantly increased the calcification of rat thoracic aorta, and the Bax/Bcl-2 ratio and Runx2 expression were up-regulated. Conclusions:Serum METTL3 level is reduced in MHD patients with VC. In vivo and in vitro studies demonstrate that METTL3 inhibits VC in CKD by mediating the apoptosis-related protein Bax/Bcl-2.

With the rapid development of pet hospitals and the increasing application of radiological diagnosis in China, the protection of staff and assistants in these hospitals is worthy of attention. Based on the progress in international management and research with respect to radiological diagnosis and protection in animal hospitals, this paper presents a brief review of the current situation, existing problems and management regulations of animal hospitals in China. It summarizes typical issues that may be encountered in radiation protection by government departments, animal hospitals, radiation professionals and assistants in animal hospitals. It also puts forward the key points of radiation protection in animal hospitals in China, which can provide scientific suggestion to assist government in decision-making and standardizing and improving the radiation protection in animal hospitals.

The 24th National Conference of Neurology of Chinese Medical Association was held in Zhuhai City, Guangdong Province during September 23-26,2021.The conference adopted a combination of online and offline methods, with a total of 2 plenary meetings, 20 special seminars, 284 invited reports, 382 papers exchanged at the conference and 1 088 papers exchanged on the wall. The conference focused on cerebrovascular diseases, epilepsy, cognitive disorders, myopathy, peripheral neuropathy, neurodegenerative diseases, nerve infectious diseases, demyelinating diseases, neuroimmune diseases, genetic and metabolic nerve diseases, nerve rehabilitation, anxiety and depression, headache, sleep disorders, nerve nursing, nerve intervention, neuroimaging, neuroelectrophysiology, translational medicine, precision medicine and other related nervous system diseases. There were more than 7 000 participants who attended this conferece.

The standardized training of specialist doctors is an important part of medical education after graduation and it is the only way to train clinicians to diagnose and treat the specialist diseases independently by using a standardized and high-quality way. The prevalence of periodontal disease and the proportion of patienets with severe and advanced periodontitis in our country are high and the diagnosis and treatment process for the periodontal disease are complicated. There is an urgent need to expand the team of periodontal specialists capable of the specialized treatment. The training of periodontal specialists in our country has just started. The present article summarizes the exploration and practice of periodontal specialist training in Peking University School and Hospital of Stomatology in the past five years, including the establishment of training bases, formulation of clear training goals, strict implementation of training rules, strengthening of process quality control to ensure the trainees could reach the expected standard of periodontal specialist after training. Through the summary of the previous stage practice, the authors hope to explore and establish a periodontal specialist training system in line with our country′s national conditions and further to promote and accelerate the pace of nationwide periodontal specialist training system.

Objective: To investigate the anti-tumor effect of CTL cells on colon cancer xenograft in nude mice after knocking out the immune check point CTLA-4 by CRISPR/Cas9 technology. Methods: A specific small guide RNA (sgRNA) for CTLA-4 was designed to construct sgRNA/Cas9 plasmid, which was then transfected into CTL using a lentiviral vector to obtain CTL cells with CTLA-4 deletion (CTLA-4 KO CTL). The transfection efficiency of the plasmid and the deletion efficiency of CTLA-4 were verified. BALB/c nude mice were randomly divided into two groups to prophylactically inoculate CTLA-4 KO CTL (experimental group) or CTL (control group); 3 days later, the animals of two groups were inoculated with colon cancer cell line LS174-T to observe the tumor formation rate and tumor formation time. After constructing colon cancer xenograft model in nude mice, the animals were randomly divided into two groups, respectively treated with CTLA-4 KO CTL (experimental group) and CTL (control group) cells to observe the tumor growth volume and survival time of mice. The serum levels of TNF-α and IFN-γ in nude mice were detected. Results: sgRNAwas designed and CRSIPR/Cas9 system with lentivirus as vector was successfully constructed. CTL cells were transfected with the established CRSIPR/ Cas9 system, and the highest transfection efficiency was up to (28.80±0.62)%. After transfection, the deletion efficiency of CTLA-4 was detected by Flow cytometry. The CTLA-4 expression of CTLA-4 KO CTL group was significantly lower than that of CTL group [(0.91±0.25)% vs (42.70±2.72)%, P<0.05]. In prophylactic assay, the formation rate of colon cancer xenografts in the experimental group was significantly lower than that in the control group(33.33%vs100%,P<0.05). In treatment assay, the tumor volume in the experimental group was significantly inhibited compared with the control group ([503±23.9] vs [911.2±51.4] mm3, P<0.05), and the survivaltimeoftheexperimentalgroupwassignificantlyprolonged (mediansurvivaltime:78dvs42d,P<0.05); Moreover, the secretion levels of serumTNF-α([268.93±17.04]pg/mlvs[148.26±20.07]pg/ml,P<0.05) and IFN-γ(315.38±18.67 pg/ml vs 202.92±29.32 pg/ml, P<0.05) in the experimental group were significantly higher than those in the control group. Conclusions: The lentiviral vector CRSIPR/Cas9 system is an effective gene editing method; its successful deletion of CTLA-4 in CTL cells can significantly inhibit the tumor formation rate of colon cancer xenografts in nude mice and enhance the anti-tumor effect of CTLon colon cancer xenografts.

Objective: To compare the efficacies of two training protocols, i.e. the multimedia instruction and the conventional method, in periodontal surgery teaching for undergraduate students. Methods: One hundred and twenty-three dental undergraduates in their pre-clinical training course were recruited and divided into two groups according to the learning grade matching principle: the experimental group (multimedia instruction, 60 students) and the control group (conventional method, 63 students). The teaching aim was to train the students gingivectomy and periodontal flap surgery by using the pig jaws. The conventional teaching method of teacher-demonstrating and student-practicing was used in the control group, and the practice time of the students′ for each surgery was 45 minutes. A standardized teaching video combined with the teachers′ explanations of the key steps was used in the experimental group. The students′ practice time for each surgery was 60 minutes. The efficacy of teaching protocol was evaluated by the teachers according to the scoring criteria set by the teaching group. Results: In gingivectomy training, accuracy of fixed points, angle of postoperative gingival margin and morphology of gingiva of the experimental group were better than those of the control group. The experimental group also had more complete and continuous excised gingivae and more thorough adjacent gingival removal. The total scores of the experimental group were significantly higher than those of the control group (92.8±2.6 vs. 89.9±3.7, P<0.05). In periodontal flap surgery training, the experimental group operated the blade around the shape of the tooth better in internal bevel incision than the control group. Additionally, the incision position of internal bevel incision, periosteal integrity after flapping and flap depth of the experimental group were better than those of the control group. Besides, the experimental group had smoother flap edge and more thorough debridement. The total scores of the experimental group were significantly higher than those of the control group (92.2±4.1 vs. 89.2±4.4, P<0.05). Conclusions The teaching efficacy of multimedia instruction was better than that of the conventional method. Its value needs to be further tested in future teaching practice.

OBJECTIVE:To understand the rationality of proton pump inhibitors (PPI) in our hospital during perioperative period,in order to provide reference for rational use of drugs in clinical practice. METHODS:Through randomly collecting discharge cases of surgical system of May and Dec. in 2015 from Hospital Information System,the use of PPI during perioperative period was investigated retrospectively,and the rationality of drug use was evaluated. RESULTS:Among 522 cases,418 cases were given PPI for preventing stress ulcer during perioperative period,with utilization rate of 80.1%. Among them,the top three were neurosurgery,orthopedics and general surgery (100%,92.6% and 92.3% respectively). Pantoprazole and lansoprazole (72.5%,14.8%,respectively) were the top two categories of the use of PPI. The irrational utilization rate of PPI was 70.8%during perioperative period. In the top three,the number of drugs was exceeded,the drug was not used,course of treatment was too long (73.7%,64.1% and 62.4% respectively). CONCLUSIONS:The utilization rate of PPI prevention is higher in the department in our hospital during perioperative period,and there is unreasonable situation in some preventive drugs,and the measures need to be implemented and strengthened.

Objective To observe the clinical efficacy of Dachaihu decoction in treatment of stroke-associated pneumonia (SAP) in the elderly patients with excess-heat syndrome and acute ischemic stroke (AIS). Methods One hundred elderly patients with excess-heat syndrome and SAP admitted to the Department of Neurology of Tianjin Nankai Hospital from August 2015 to September 2016 were enrolled, and they were divided into a control group and an observation group according to the random number table, 50 cases in each group. Both groups were given conventional neurological treatment, while the observation group additionally received Dachaihu decoction (the ingredients of the decoction: bupleurum 15 g, scutellariae 10 g, pinellia 10 g, fructus aurantii immaturus 15 g, radix paeoniae alba 10 g, rhubarb 5 g, ginger 10 g, jujube 2), and according to the individual's symptoms and signs, the above ingredients can be added and/or subtracted, 1 dose per day, divided into two parts, 1 part taken in the morning and 1 part in the evening orally, 14 days as a therapeutic course. The body temperature recovery time, the duration of antibiotic treatment, the changes of Clinical Pulmonary Infection Score (CPIS) and National Institutes of Health Stroke Scale (NIHSS) score before and after treatment of the two groups were observed and the therapeutic effects in the two groups were evaluated. Results The body temperature recovery time (days: 6.56±2.70 vs. 8.08±3.23) and the duration of antibiotic treatment (days: 8.78±2.80 vs. 10.02±2.90) in the observation group were significantly shorter than those in the control group (both P ＜ 0.05); with the prolongation of treatment, after treatment, the CPIS and NIHSS score were decreased significantly in both groups, reaching the lowest levels on the 14th day, and the degrees of score descent in observation group were more significant than those of the control group (CPIS score: 0.68±0.59 vs. 1.12±0.96, NIHSS score: 6.38±2.31 vs. 7.44±2.74, both P ＜ 0.05); the total effective rate in the observation group was significantly higher than that in the control group [92% (46/50) vs. 76% (38/50), P ＜ 0.05]. Conclusion The therapeutic effect of Dachaihu decoction used for treatment of SAP in elderly patients with excess-heat syndrome is favorably effective and safe.