Macroautophagy (referred to as autophagy hereafter) is a major intracellular lysosomal degradation pathway that is responsible for the degradation of misfolded/damaged proteins and organelles. Previous studies showed that autophagy protects against acetaminophen (APAP)-induced injury (AILI) via selective removal of damaged mitochondria and APAP protein adducts. The lysosome is a critical organelle sitting at the end stage of autophagy for autophagic degradation via fusion with autophagosomes. In the present study, we showed that transcription factor EB (TFEB), a master transcription factor for lysosomal biogenesis, was impaired by APAP resulting in decreased lysosomal biogenesis in mouse livers. Genetic loss-of and gain-of function of hepatic TFEB exacerbated or protected against AILI, respectively. Mechanistically, overexpression of TFEB increased clearance of APAP protein adducts and mitochondria biogenesis as well as SQSTM1/p62-dependent non-canonical nuclear factor erythroid 2-related factor 2 (NRF2) activation to protect against AILI. We also performed an unbiased cell-based imaging high-throughput chemical screening on TFEB and identified a group of TFEB agonists. Among these agonists, salinomycin, an anticoccidial and antibacterial agent, activated TFEB and protected against AILI in mice. In conclusion, genetic and pharmacological activating TFEB may be a promising approach for protecting against AILI.

Purpose To investigate the clinicopathological features,diagnosis and differential diagnosis of the primordial odontogenic tumour(POT).Methods Clinical data of 4 cases of jawbone POT were collected.Imaging examination,HE,and immunohistochemical EnVision two-step staining was used to an-alyze their clinical and pathological characteristics,and relevant literatures were reviewed.Results The age arranged from 5 years to 21 years.2 cases were male and 2 case were female.There were 2 cases in maxilla and 2 cases in mandible.The clinical presentation was a slow growing painless mass.Cut sur-face of the tumor was appeared grayish yellow and grayish white,the tumor involved the crown of an unerupted tooth.The tumour consisted of a proliferation of spindled and stellate cells in myx-oid stroma.Surfaced by cuboidal to columnar epithelium forming papillary structures and invaginations.Calcification was observed in 2 cases.Conclusion POT is a rare benign mixed odontogen-ic tumor that is more common in children and adolescents.Mas-tering its characteristic histological morphology can make a cor-rect diagnosis.Local complete resection of the tumor has a good prognosis.

Objective:To study the molecular mechanism of functional defect of protein C (PC) caused by point mutations of human protein C gene (PROC) N355S,G392E and T314A. Methods:The wild-type and mutant plasmids (PCWT,PCN355S,PCG392E,PCT314A) of PROC gene were constructed and transiently transfected into HEK293 cells. The expression of mutant proteins in vitro were tested. The mRNA level changes of wild-type and mutant PC after 24 h of transfection were detected by real-time PCR. Western blot and ELISA were used to detect the changes of intracellular and extracellular protein levels of wild-type and mutant PC. The supernatant of cells transfected for 24-48 h was concentrated by ultrafiltration. The protein in the concentrated solution was quantified,and PC activation and enzyme kinetics tests were performed. Clustal Omega multiple sequence alignment was used to analyze the conservation of amino acid mutation sites. The effect of mutation on PC protein structure was analyzed by PyMOL software. Results:The relative expression abundances of PROC mRNA in PCN355S,PCG392E and PCT314A groups were 1.14±0.46,0.96±0.08 and 1.08±0.17,respectively,and there were no significant differences compared with 1.02±0.24 in PCWT group (P>0.05). Western blot analysis of the lysates of transfected cells showed that the content of PCT314A recombinant protein slightly decreased and the band became relatively lighter. The ELISA results of the concentrated cell culture supernatants showed that the PC:Ag levels of PCN355S and PCG392E were 98.8％±2.4％ and 101.4％±3.1％,respectively,with no significant differences compared with PCWT,while PCT314A decreased compared with PCWT (PC:Ag:88.6％±3.2％) (P<0.05). The results of enzyme kinetics test showed that APCN355S (Km=338.3±43.2,Vmax=2.015±0.12),APCG392E (Km=292.2±28.4,Vmax=1.893±0.07) and APCT314A (Km=299.5±24.6,Vmax=1.775±0.06) showed an increase in Km and a decrease in Vmax compared with APCWT (Km=238.2±4.58,Vmax=3.205±0.06). Multiple sequence alignment suggested that the three mutations be highly conservative in different species. The structural model suggested that the amino acid substitutions of N355S,G392E and T314A mutations collide with the surrounding amino acid groups,causing distortion of the surrounding structure,which may have adverse effects on the folding and biological function of PC. Conclusion:The N355S,G392E and T314A mutations in the PROC gene cause functional defects in PC by weakening the binding between PC and substrate. These three mutations have caused serious spatial collisions in the protein structure,affecting the folding of PC and the reactivity of active sites.

Objective:To analyze the epidemiological and clinical characteristics of patients with brucellosis.Methods:Medical records of confirmed brucellosis patients ( n = 101) were collected from January 2015 to December 2022 at the First Affiliated Hospital of Nanjing Medical University. Patients were divided into acute phase group (≤3 months, n = 89) and non acute phase group (> 3 months, n = 12) according to the course of the disease. The epidemiological characteristics, clinical symptoms, laboratory indicators, treatment plan and curative effect of the patients were analyzed retrospectively. Results:Data of a total of 101 cases of brucellosis were collected, including 72 males and 29 females. The disease occurred throughout the year, with summer (June to August) being the peak period (43.56%, 44/101); 72.28% (73/101) cases had a clear history of contact with animal. Ninety-two point zero eight percent (93/101) of the patients visited the Department of Infectious Diseases for the first time. Clinical manifestations included fever, accounting for 82.18% (83/101), chills accounting for 36.63% (37/101), backache accounting for 33.66% (34/101), night sweats accounting for 22.77% (23/101), and arthralgia accounting for 20.79% (21/101). The symptoms of backache (75.00%, 9/12) and arthralgia (41.67%, 5/12) were more common in patients in the non acute phase group than those of the acute phase group [28.09% (25/89), 17.98% (16/89), P < 0.05]. The most common laboratory test abnormal items were elevated C-reactive protein (CRP, 68.32%, 69/101), erythrocyte sedimentation rate (ESR, 61.39%, 62/101), aspartate aminotransferase (AST, 58.42%, 59/101), and alanine aminotransferase (ALT, 48.51%, 49/101). Elevated ESR (66.29%, 59/89) was more common in patients in the acute phase group than that of the non acute phase group [25.00 (3/12), χ 2 = 7.48, P = 0.006]. All patients were treated with a combination therapy, with a recovery rate of 100.00% (101/101). Conclusions:Brucellosis patients are more common in males, with a higher incidence in summer and often accompanied by a history of contact with animal. The clinical manifestations are diverse and non-specific.

The nucleus accumbens (NAc) plays an important role in various emotional and motivational behaviors that rely on heightened wakefulness. However, the neural mechanisms underlying the relationship between arousal and emotion regulation in NAc remain unclear. Here, we investigated the roles of a specific subset of inhibitory corticotropin-releasing hormone neurons in the NAc (NAc^CRH) in regulating arousal and emotional behaviors in mice. We found an increased activity of NAc^CRH neurons during wakefulness and rewarding stimulation. Activation of NAc^CRH neurons converts NREM or REM sleep to wakefulness, while inhibition of these neurons attenuates wakefulness. Remarkably, activation of NAc^CRH neurons induces a place preference response (PPR) and decreased basal anxiety level, whereas their inactivation induces a place aversion response and anxious state. NAc^CRH neurons are identified as the major NAc projection neurons to the bed nucleus of the stria terminalis (BNST). Furthermore, activation of the NAc^CRH-BNST pathway similarly induced wakefulness and positive emotional behaviors. Taken together, we identified a basal forebrain CRH pathway that promotes the arousal associated with positive affective states.
Animals ; Septal Nuclei/metabolism* ; Nucleus Accumbens/physiology* ; Corticotropin-Releasing Hormone/metabolism* ; Wakefulness/physiology* ; Neurons/metabolism* ; Male ; Mice ; Mice, Inbred C57BL ; Neural Pathways/physiology* ; Anxiety/physiopathology* ; Reward

Humans ; Uric Acid ; Cross-Sectional Studies ; Spondylarthritis/diagnostic imaging* ; C-Reactive Protein ; Spondylitis, Ankylosing

OBJECTIVE: To observe the effects of Xingnao Kaiqiao (regaining consciousness and opening orifices) acupuncture therapy on the expression of hypoxia-inducible factor 1α (HIF-1α) and Nod-like receptor protein 3 (NLRP3) in cerebral ischemia-reperfusion rats, and to explore the mechanism of acupuncture against cerebral ischemia-reperfusion injury. METHODS: Seventy-two male SD rats were randomly divided into a sham-operation group, a model group, an acupuncture group and a non-point acupuncture group, with 18 rats in each one. Using modified Longa thread embolization method, the rat model of acute focal cerebral ischemia was prepared; and after 2 h ischemia, the reperfusion was performed to prepared the model of cerebral ischemia-reperfusion. Immediately after reperfusion, Xingnao Kaiqiao acupuncture method was applied to bilateral "Neiguan" (PC 6) and "Shuigou" (GV 26) in the acupuncture group, while in the non-point acupuncture group, acupuncture was delivered at non-points and all of the needles were retained for 30 min in these two groups. The samples were collected 24 h after reperfusion in the rats of each group. Zea-Longa neurological deficit score was used to evaluate the degree of cerebral neurological impairment, TTC staining was adopted to observe the volume percentage of cerebral infarction, HE staining was provided to observe the morphological changes of brain, and Western blot was applied for detecting the expression of HIF-1α and NLRP3 proteins in the cerebral cortex on the right side. RESULTS: Compared with the sham-operation group, neurological deficit score and volume percentage of cerebral infarction were increased in the model group (P<0.01), and HIF-1α and NLRP3 protein expression was elevated (P<0.01). Compared with the model group, neurological deficit score and volume percentage of cerebral infarction were decreased (P<0.01), and HIF-1α and NLRP3 protein expression was lower (P<0.01) in the acupuncture group. There was no significant difference in above indexes in the non-point acupuncture group compared with the model group (P>0.05). Compared with the sham-operation group, the brain tissue of the rats in the model group and the non-point acupuncture group was loose and edema, and the nuclei were shriveled. The brain tissue morphology in the acupuncture group was similar to that of the sham-operation group. CONCLUSION Acupuncture can alleviate cerebral ischemia-reperfusion injury, and its mechanism may be related to the regulation of HIF-1α/NLRP3 signaling pathway to attenuate inflammatory response.
Male ; Animals ; Rats ; Rats, Sprague-Dawley ; NLR Family, Pyrin Domain-Containing 3 Protein/genetics* ; Acupuncture Therapy ; Reperfusion Injury/therapy* ; Brain Ischemia/therapy* ; Cerebral Infarction/therapy* ; NLR Proteins

As the body ages， the immune system will undergo a series of changes， which are termed "immunosenescence" and are embodied in immune cells. Previous studies have shown that the immune cells involved in the regulation of immunosenescence include intrinsic immune cells and adaptive immune cells. Intrinsic immune cells are neutrophils， monocytes/macrophages， myeloid-derived suppressor cells， dendritic cells， natural killer cells， etc.， and the underlying mechanisms involve the regulation of cell number， phagocytosis， chemotaxis， adhesion， the function of toll-like receptor （TLR）， antigen presentation， macrophage polarization， cytotoxicity， migration， etc. The adaptive immune cells include T-lymphocytes and B-lymphocytes， and the underlying mechanisms involve the regulation of cell development， proliferation， differentiation， cell number， telomerase activity， self-reactive antibodies， etc. Immunosenescence is the manifestation of aging in the human body and is also an important target for delaying aging by Chinese medicine and western medicine. In recent years， scholars have found some classical prescriptions and their active components （such as Dushentang and total saponins in Panax ginseng leaves， and Shengmaiyin and anwulignan and total saponins in P. ginseng stems and leaves） can regulate immunosenescence by targeting the immune cells and interfering with their molecular regulatory mechanisms. In addition， the mechanisms of the classical prescriptions in regulating immunosenescence are closely related to autophagy. The representative prescription embodying the therapeutic principles of resolving blood stasis and promoting regeneration， Dahuang Zhechongwan， can delay D-galactose-induced renal aging in mice， and its underlying mechanisms are related to the regulation of the number and activity of thymic immune cells and improvement of the protein expression of autophagy-related markers and inflammatory cytokines in the kidney. Therefore， exploring the effects of the classical prescriptions and their active components by targeting the mechanisms of immunosenescence will become a new direction for investigating and developing anti-aging drugs.

Objective:To investigate the distribution characteristics of bacteria in urine of patients with ureteral stent crusting.Methods:Thirty-five patients who underwent ureteral stent placement at the Shandong Provincial Third Hospital, Shandong University Qilu Hospital, Jinan Central Hospital, and Jinan Jigang Hospital were selected from October, 2018 to March, 2019(the clinical study registration number is ChiCTR1800020025). The inclusion criteria were patients who had the stent intubated for 4 weeks after ureteroscopic lithotripsy, aged between 18 and 65 years. Exclusion criteria were patients with positive urine bacterial culture, severe gross hematuria, recent oral antibiotics, and patients with significant residual stones. This clinical study uses a cross-sectional study method, and those patients were divided into crusting group (n=23) and non-crusting group (n=12) according to the presence or absence of stent crusting. On the day of extubation, urine of the patients was collected for bacterial 16s DNA detection. The distribution characteristics of bacteria in urine of the two groups were analyzed using UPARSE, UCHIME and RDP calssifier. The total number of bacteria species, bacterial abundance and bacterial species with large-scale abundance in urine of the two groups were determined. The quantity of bacteria species and bacterial abundance in the urine between the two groups were compared, and the bacterial species with large-scale abundance in urine of the patients with stent crusting were identified.Results:There were no significant differences in general information such as age, body mass index, gender, affected side, type of stent tube, and stone composition between the two groups. Using 16s DNA sequencing to detect the bacteria in the urine of the two groups revealed that the number of bacterial species with abundance >1% was 11, and the number of bacterial species with abundance >0.01% was 74 in the crusting group. In the non-crusting group, the number of bacterial species with abundance >1% and >0.01% was 7 and 11, respectively. Compared with the non-crusting group, the number of bacterial species with abundance >1% in the crusting group was significantly larger ( t=5.12, P=0.000). In the crusting group, bacterial species with the top three abundance were g_Lactobacillus (23.1%), g_Bacteroides (18.8%) and g_norank_Bacteroides (17.1%). In the non-crusting group, bacterial species with the top three abundance were g_Escherichia-Shigella (32.2%), g_Enterococcus (24.9%) and g_Pseudomonas (18.2%). The three bacteria with the greatest difference between the two groups were g_ Lactobacillus ( P=0.010), g_Bacteroides ( P=0.004) and g_norank_Bacteroides ( P=0.004), respectively. Conclusion:The species and quantity of bacteria in the urine of patients with stent crusting are both significantly larger than those of patients without stent crusting. Bacteroides with larger-scale abundance in the urine of patients with stent crusting may promote the deposition of crystals on the stent wall through its structure, function and urease positive characteristics.

COVID-19, an infectious disease caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), has spread throughout the world. China has achieved rapid containment of this highly infectious disease following the principles of early detection, early quarantine and early treatment with integrated traditional Chinese and Western medicine. The inclusion of traditional Chinese medicine (TCM) in the Chinese protocol is based on its successful historic experience in fighting against pestilence. Current findings have shown that the Chinese medicine can reduce the incidence of severe or critical events, improve clinical recovery and help alleviate symptoms such as cough or fever. To date there are over 133 ongoing registered clinical studies on TCM/integrated traditional Chinese and Western medicine. The three Chinese patent medicines (/ (Forsythiae and Honeysuckle Flower Pestilence-Clearing Granules/Capsules), (Honeysuckle Flower Cold-Relieving Granules) and (Stasis-Resolving & Toxin-Removing) were officially approved by the National Medical Products Administration to list COVID-19 as an additional indication. The pharmacological studies have suggested that Chinese medicine is effective for COVID-19 probably through its host-directed regulation and certain antiviral effects.