1.Chaihu Longgu Mulitang Relieves Generalized Anxiety Disorder in Rats via p38 MAPK/NF-κB Signaling Pathway
Chunxin WEI ; Yiming HU ; Shiqi HUANG ; Guowei TAN ; Yaorong AN
Chinese Journal of Experimental Traditional Medical Formulae 2024;30(4):114-123
ObjectiveTo study whether Chaihu Longgu Mulitang can inhibit hypothalamic inflammation, mitigate anxiety-like behavior, and alleviate anxiety symptoms by regulating the p38 mitogen-activated protein kinase/nuclear factor-κB (p38 MAPK/NF-κB) signaling pathway in the rat model of generalized anxiety disorder (GAD). MethodTwelve out of 74 Wistar rats were randomly selected as the blank group, and the remaining rats were subjected to chronic restraint stress for the modeling of GAD. The open field test (OFT) and elevated Porteus maze test (PMT) were conducted 14 days after modeling to detect the anxiety-like behaviors. Sixty successfully modeled rats were selected and randomized into model, low-, medium-, and high-dose (6, 12, and 24 g·kg-1, respectively) Chaihu Longgu Mulitang, and diazepam (1 mg·kg-1) groups (n=12) and administrated with corresponding drugs for 14 consecutive days. OFT and PMT were then carried out to examine the anxiety-like behaviors of the rats. The levels of tumor necrosis factor-α (TNF-α), interleukin-6 (IL-6), and interleukin-1β (IL-1β) in the hypothalamus and serum of rats were determined by the enzyme-linked immunosorbent assay (ELISA). Real-time fluorescence quantitative polymerase chain reaction (Real-time PCR)was conducted to determine the mRNA levels of p38 MAPK, NF-κB p65, nuclear factor κB inhibitor α (IκBα), and ionized calcium binding adaptor molecule 1 (Iba-1). The protein levels of p38 MAPK, phosphorylated (p)-p38 MAPK, NF-κB p65, p-NF-κB p65, and IκBα in the hypothalamus of rats were determined by Western blot. The expression of Iba-1 in the hypothalamic microglia was detected by immunofluorescence assay. ResultCompared with the blank group, the model group had decreased body weight, scattered dark yellow fur, increased irritability, and preference to hibernation in the corner. In addition, the modeled rats showed increased edge movement distance and time in OFT (P<0.01) and decreased movement distance and time and the number of entries in the open arm in PMT (P<0.01). The modeling increased the fluorescence intensity of Iba-1 in paraventricular nucleus of hypothalamus (P<0.01), elevated the levels of IL-1β, IL-6, and TNF-α in the serum and hypothalamus (P<0.01), up-regulated the protein and mRNA levels of p38 MAPK, NF-κB p65, p-p38 MAPK, p-NF-κB p65, and Iba-1 (P<0.05, P<0.01), and down-regulated the protein and mRNA levels of IκBα (P<0.01) in the hypothalamus. Compared with the model group, medium- and high-dose Chaihu Longgu Mulitang and diazepam increased the body weight, improved the fur and behaviors, decreased the edge movement distance and time in OFT (P<0.05, P<0.01), and increased the movement distance and time in the open arm in PMT (P<0.05, P<0.01). Furthermore, they decreased the fluorescence intensity of Iba-1 in hypothalamic microglia (P<0.05, P<0.01), lowered the levels of IL-1β, IL-6, and TNF-α in the serum and hypothalamic tissue (P<0.05, P<0.01), down-regulated the mRNA and protein levels of p38 MAPK, NF-κB p65, p-p38 MAPK, p-NF-κB p65, and Iba-1 (P<0.05, P<0.01), and up-regulated the mRNA and protein levels of IκBα (P<0.05, P<0.01) in the hypothalamus. ConclusionChaihu Longgu Mulitang can mitigate anxiety-like behaviors and relieve anxiety in GAD rats by inhibiting the p38 MAPK/NF-κB signaling pathway and reducing the activation of microglia and the levels of pro-inflammatory cytokines in the hypothalamus.
2.Tubeless three-port versus conventional single utility port thoracoscopic surgery in the treatment of thymic tumors: A retrospective cohort study
Yuchi XIU ; Bo LIU ; Hao MENG ; Renquan DING ; Xingchi LIU ; Shiqi WANG ; Boxiao HU ; Qiong WU ; Guoqing ZHANG ; Shiguang XU
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2024;31(04):546-550
Objective To investigate the safety and efficacy of totally no tube three-port thoracoscopic surgery (TNTT) for thymic tumor via lateral thoracic approach. Methods The clinical data of patients with thymoma admitted to the Department of Thoracic Surgery of the General Hospital of Northern Theater Command from November 2021 to May 2022 were retrospectively analyzed. The patients were divided into a TNTT group and a single utility port video-assisted thoracic surgery (SVATS) group according to different surgical methods. The clinical data were compared between the two groups. Results A total of 111 patients were collected. There were 44 patients in the TNTT group, including 20 males and 24 females, with an average age of 60.11±8.64 years, and 67 patients in the SVATS group, including 30 males and 37 females, with an average age of 62.40±7.92 years. There was no significant difference between the two groups in the baseline data (P>0.05). The postoperative hospital stay and intraoperative blood loss were shorter or less in the TNTT group (P<0.05), and the visual analogue scale score 48 hours after the operation was smaller in the SVATS group (P<0.05). Conclusion TNTT has a good surgical safety, and can shorten postoperative hospital stay, reduce intraoperative blood loss, and has significant advantages in enhanced recovery after surgery, but SVATS can reduce postoperative pain in patients.
3.Discussion of factors influencing preclinical studies based on fecal bacteria transplantation in mice
Shiqi SUN ; Lu LIU ; Shuangyuan HU ; Yuyan WANG ; Mingsheng SUN ; Ling ZHAO
Acta Laboratorium Animalis Scientia Sinica 2024;32(1):100-117
Fecal microbiota transplantation(FMT)is a therapeutic approach that targets intestinal microorganisms by transplanting fecal microorganisms from healthy individuals into the gastrointestinal tract of diseased individuals,thus restoring the recipient's disordered gastrointestinal microbiota by restructuring the intestinal flora.However,the mechanism of action and adverse effects of FMT in different diseases have not yet been clarified,thus limiting its wide clinical application.Its use still relies on in-depth preclinical studies;however,highly inconsistent or incomplete experimental details provided in current reports,coupled with a lack of authoritative standards and recommendations,seriously affect the interpretation of the study findings and replication of the experimental procedures,as well as hindering the clinical translation of the result.We therefore review and discuss the key steps of recipient selection and graft sample collection,storage,graft material preparation,and grafting route,with the aim of improving the utilization of experimental animals,consumables,and labor,and providing method ological recommendations and references to achieve replicability and standardization of preclinical FMT studies.
4.Imaging features of peripapillary hyperreflective ovoid mass-like structure
Jing HU ; Lu HE ; Shiqi TANG
Chinese Journal of Experimental Ophthalmology 2024;42(9):858-861
Peripapillary hyperreflective ovoid mass-like structure (PHOMS) is a novel imaging feature newly defined in recent years in the study of optic disc drusen (ODD).In OCT images, it shows an oval hyper-reflective signal on the Bruch membrane around the optic papilla, ring or crescent non-fluorescence in autofluorescence, and light gray peripapillary elevations around the optic disc in non-red infrared images.There is no conclusive evidence as to whether PHOMS is a subtype or precursor of ODD.Histopathologically, PHOMS might correspond to the lateral bulging or herniation of distended axons into the peripapillary retina.Since there may be different pathological properties between PHOMS and ODD in histology, there are different imaging features in multimodal imaging.To make the diagnosis of ODD more accurate, the Optic Disc Drusen Studies Consortium recommends that PHOMS be distinguished from ODD.PHOMS is mainly differentiated from ODD and perioptic vessels on OCT.It can be seen in many diseases such as ODD, optic papilledema, non-arteritis anterior ischemic optic neuropathy, multiple sclerosis, central retinal vein occlusion, pediatric tilted disc syndrome, isolated optic neuritis and optic atrophy, and is an important cause of pseudopapilloedema in children.This article summarizes the origin, imaging features, differential diagnosis and related diseases of PHOMS, to increase domestic researchers' understanding of the structure of PHOMS, enhance the differential diagnosis between PHOMS and ODD, and provide a reference for clinical research and diagnosis.
5.Expression and significance of ubiquitin-specific proteases 20 and hypoxia inducible factor-1α in breast cancer
Lingyu FANG ; Jinghua HU ; Junfeng WEN ; Shiqi HAN ; Yali WANG ; Lulan PU ; Jingjia LI ; Yi YANG ; Shishan DENG ; Lingmi HOU ; Fangfang ZHOU
The Journal of Practical Medicine 2024;40(16):2270-2276
Objective To explore the changes and significane of USP20 and HIF-α expression in breast cancer.Methods Following transfection of shRNA-USP20 lentivirus into breast cancer MDA-MB-231 cells,the gene and protein expression levels of USP20 were detected using fluorescence quantitative PCR and Western Blot.Subsequently,the overexpression of USP20 was observed to determine its effect on HIF-α expression.Similarly,siRNA-USP20 was used to knock down USP20 in breast cancer MDA-MB-231 cells,followed by detection of gene and protein expression levels using fluorescence quantitative PCR and Western Blot.The subsequent changes in HIF-α expression were then examined.Rusults The positive expression rates of USP20 and HIF-α in breast cancer tissues were 69.6%and 46.83%,respectively,while they were negatively expressed in the adjacent normal tissues,with statistically significant differences(P<0.01).The positive expressions of USP20 and HIF-α were predomi-nantly observed in the cytoplasm of breast cancer tissue,with a smaller amount present in the nucleus.There was a significant positive correlation between USP20 and HIF-α in breast cancer.Following transfection of shRNA-USP20 lentivirus into MDA-MB-231 cells,both the protein and gene expression levels of USP20 significantly increased(P<0.01).Over-expression of USP20 did not affect HIF-α mRNA levels but led to a significant increase in HIF-α protein expression(P<0.01).Conversely,siRNA-USP20 interference resulted in a significant decrease in both the protein and gene expression levels of USP20(P<0.01),without affecting HIF-α mRNA levels;however,it caused a notable reduction in HIF-α protein expression(P<0.01).Conclusion The expression of USP20 exhib-ited a significant positive correlation with HIF-α in breast cancer.Overexpression of USP20 led to a substantial increase in HIF-α protein expression,while knock-down of the USP20 gene resulted in a significant decrease in HIF-α protein levels.Therefore,it can be inferred that USP20 may exert its influence on the development of breast cancer through modulation of HIF-α expression,thereby providing crucial experimental evidence for clinical treat-ment,prognosis,and further investigations.
6.Evaluation of the humoral immunity in mice induced by foot-and-mouth disease virus-like particles-ZIF-8 complexes with different sizes.
Jiajun LI ; Jun WANG ; Yun ZHANG ; Zhidong TENG ; Hu DONG ; Huichen GUO ; Shiqi SUN
Chinese Journal of Biotechnology 2023;39(12):4837-4848
To further enhance the immune effect of the foot-and-mouth disease (FMD) virus-like particles (VLPs) vaccine, this study prepared FMDV VLPs-zeolitic imidazolate (framework-8, ZIF-8) complexes with different particle sizes. We used a biomimetic mineralization method with Zn2+ and 2-methylimidazole in different concentration ratios to investigate the effect of size on the immunization effect. The results showed that FMDV VLPs-ZIF-8 with three different sizes were successfully prepared, with an approximate size of 70 nm, 100 nm, and 1 000 nm, respectively. Cytotoxicity and animal toxicity tests showed that all three complexes exhibited excellent biological safety. Immunization tests in mice showed that all three complexes enhanced the titers of neutralizing and specific antibodies, and their immune effects improved as the size of the complexes decreased. This study showed that ZIF-8 encapsulation of FMDV VLPs significantly enhanced their immunogenic effect in a size-dependent manner.
Animals
;
Mice
;
Foot-and-Mouth Disease/prevention & control*
;
Foot-and-Mouth Disease Virus
;
Antibodies, Neutralizing
;
Immunity, Humoral
;
Immunization
;
Vaccines, Virus-Like Particle
;
Antibodies, Viral
;
Viral Vaccines
7.Construction of foot-and-mouth disease virus like particles-induced expression vectors and screening of BHK-21 cell pools.
Shuzhen TAN ; Hu DONG ; Shiqi SUN ; Huichen GUO
Chinese Journal of Biotechnology 2023;39(12):4849-4860
Transient expression is the major method to express foot-and-mouth disease virus (FMDV) capsid proteins in mammalian cells. To achieve stable expression of FMDV capsid proteins and efficient assembly of virus like particles (VLPs) in cells, the plasmids of piggyBac (PB) transposon-constitutive expression and PB transposon-tetracycline (Tet) inducible expression vectors were constructed. The function of the plasmids was tested by fluorescent proteins. By adding antibiotics, the constitutive cell pools (C-WT, C-L127P) expressing P12A3C (WT/L127P) genes and the inducible cell pools (I-WT, I-L127P) expressing P12A3C (WT/L127P) genes were generated. The genes of green fluorescent protein, 3C protease and reverse tetracycline transactivator (rtTA) were integrated into chromosome, which was confirmed by fluorescence observation and PCR testing. The cell pool I-L127P has a stronger production capacity of capsid proteins and VLPs, which was confirmed by Western blotting and enzyme linked immunosorbent assay (ELISA), respectively. In conclusion, inducing the chromosomal expression of FMDV capsid proteins was firstly reported, which may facilitate the technical process of mammalian production of FMDV VLPs vaccine and the construction of mammalian inducible expression systems for other proteins.
Animals
;
Foot-and-Mouth Disease Virus/genetics*
;
Capsid Proteins
;
Viral Proteins/metabolism*
;
Foot-and-Mouth Disease/prevention & control*
;
Tetracyclines/metabolism*
;
Viral Vaccines
;
Antibodies, Viral
;
Mammals/metabolism*
8.Research Ideas and Challenge of Real World Study and Artificial Intelligence Based On Clinical Diagnosis and Treatment Data of Traditional Chinese Medicine
Guozhen ZHAO ; Shiqi GUO ; Huaxin PANG ; Ziheng GAO ; Bo LI ; Zhaolun CAI ; Shiyan YAN ; Dongran HAN ; Yixing LIU ; Jing HU ; Qingquan LIU
Journal of Traditional Chinese Medicine 2023;64(21):2170-2175
With the continuous progress of research methodology in the real world and the growing maturity of artificial intelligence technology, a method for conducting “quantitative” research to guide clinical practice based on traditional Chinese medicine (TCM) diagnosis and treatment data was gradually developed. However, there is still a need for further improvements in the overall design of studies and the transformation of findings into clinical practice. Based on this, we put forward a comprehensive overall design concept and application approach for real-world study and artificial intelligence research based on clinical diagnosis and treatment data of TCM. This approach consists of five steps: Constructing a research-based database with a large sample size and high data quality; Mining and classification of core prescriptions; Conducting cohort studies to evaluate the effectiveness of core prescriptions; Utilizing case-control studies to clarify the dominant population; Establishing predictive models to achieve precision medicine. Additionally, it is imperative for researchers to establish a standardized system for collecting TCM variables and processing data, optimize the determination and measurement methods of confounding factors, further improve and promote methodologies, and strengthen the training of interdisciplinary talents. By following this research method, we anticipate that the clinical translation of research findings will be facilitated, leading to advancements in TCM precision medicine. Real-world study and artificial intelligence research share similar research foundations, and clinical applications complement each other. In the future, the two will merge together.
9.Effects of nimodipine on cognitive function in patients with Alzheimer's disease
Weiwei HU ; Wenke WANG ; Shiqi YANG
Chinese Journal of Primary Medicine and Pharmacy 2023;30(4):514-517
Objective:To investigate the effects of nimodipine on cognitive function in patients with Alzheimer's disease.Methods:A total of 200 patients with Alzheimer's disease who received treatment in Shuangqiao Hospital from January 2019 to January 2022 were included in this study. They were randomly divided into a control group and an observation group ( n = 100/group). The control group was treated with Donepezil. The observation group was treated with nimodipine and Donepezil. Before and 2 months after treatment, mental status and cognitive function were evaluated in each group. Results:After treatment, total response rate in the observation group was significantly higher than that in the control group [95% (95/100) vs. 81% (81/100), χ2 = 9.58, P < 0.05]. Alzheimer's disease assessment scale cognitive subscale score was significantly lower after treatment compared with before treatment in each group ( P < 0.05). Mini-mental State Examination score was significantly higher after treatment compared with before treatment in each group ( P < 0.05). Alzheimer's disease assessment scale cognitive subscale score in the observation group was significantly lower than that in the control group [(17.38 ± 1.95) points vs. (29.63 ± 3.39) points, t = -3.26, P < 0.05]. Mini-mental State Examination score in the observation group was significantly higher than that in the control group [(23.47 ± 4.59) points vs. (18.68 ± 3.91) points, t = 2.14, P < 0.05]. Conclusion:Nimodipine can improve cognitive function in patients with Alzheimer's disease.
10.Risk factors for postoperative chylothorax after robot-assisted versus video-assisted thoracic surgery in radical lung cancer resection: A propensity score matching study
Deyu LIU ; Shiguang XU ; Wei XU ; Bo LIU ; Bo LI ; Xilong WANG ; Boxiao HU ; Shiqi WANG ; Yuchi XIU ; Shumin WANG
Chinese Journal of Clinical Thoracic and Cardiovascular Surgery 2022;29(04):449-456
Objective To compare the postoperative chylothorax outcomes of robot-assisted thoracic surgery (RATS) and video-assisted thoracoscopic surgery (VATS), analyze the risk factors for postoperative chylothorax after minimally invasive radical lung cancer resection and explore possible prevention and control measures. Methods Between June 2012 and September 2020, 1 083 patients underwent minimally invasive pulmonary lobectomy and systematic lymph node dissection in our hospital, including 578 males and 505 females with an average age of 60.6±9.4 years. Patients were divided into two groups according to the operation methods: a RATS group (499 patients) and a VATS group (584 patients). After propensity score matching, 434 patients were included in each group (868 patients in total). Chylothorax and other perioperative indicators were compared between the two groups. Univariate and multivariate logistic regression analyses were performed to identify risk factors for postoperative chylothorax. Results Overall, 24 patients were diagnosed with chylothorax after surgery. Compared with the VATS group, the rate of chylothorax was higher (3.9% vs. 1.6%, P=0.038), the groups and numbers of dissected lymph nodes were more (both P<0.001), and the intraoperative blood loss was significantly less (P<0.001) in the RATS group. There was no statistical difference in the postoperative hospital stay (P=0.256) or chest tube drainage time (P=0.504) between the two groups. Univariate analysis showed that gender (P=0.021), operation approach (P=0.045), smoking (P=0.001) and the groups of dissected lymph nodes (P<0.001) were significantly associated with the development of chylothorax. Multivariate analysis showed that smoking [OR=4.344, 95%CI (1.149, 16.417), P=0.030] and the groups of dissected lymph nodes [OR=1.680, 95%CI (1.221, 2.311), P=0.001] were the independent risk factors for postoperative chylothorax. Conclusion Compared with the VATS, the rate of chylothorax after RATS is higher with more dissected lymph nodes and less blood loss. The incidence of chylothorax after minimally invasive radical lung cancer resection is higher in the patients with increased dissected lymph node groups and smoking history.

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