ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis， evaluate its potential clinical application value， and compare the therapeutic effect with that of Cranial Painkiller granules， so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis， and the rats were randomly divided into the normal group， model group， carbamazepine group （60 mg·kg^-1）， Cranial Painkiller granules group （2.70 g·kg^-1）， and low， medium， and high dosage groups of Cranial Painkiller pills' extract powder （1.35， 2.70， 5.40 g·kg^-1） according to the basal mechanical pain thresholds， and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling， and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin （HE） staining was used to detect pathological changes in the trigeminal ganglion， and enzyme-linked immunosorbent assay （ELISA） was used to detect the inflammatory factors interleukin-1 （IL-1）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， and tumor necrosis factor-α （TNF-α） levels in rat serum， as well as neuropeptide substance P （SP） and β-endorphin （β-EP） levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3， ASC， Caspase-1， and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group， the pain threshold of rats in the model group showed a continuous significant decrease （P<0.01）. The pathological damage of brain tissue was significant （P<0.01）， and the inflammatory levels of IL-1， IL-6， IL-8， and TNF-α in serum were significantly elevated （P<0.01）. The level of the SP in the brain tissue was significantly elevated （P<0.01）， and the level of β-EP was significantly reduced （P<0.01）， while the level of OTULIN was significantly reduced， and NLRP3， ASC， and Caspase-1 protein levels were significantly elevated （P<0.01）. After administration of the drug， compared with the model group， the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased （P<0.01）. The inflammatory levels of IL-1， IL-6， IL-8， and TNF-α and SP levels significantly decreased （P<0.01）， and the β-EP levels were significantly elevated （P<0.01）， while the levels of OTULIN protein were significantly elevated （P<0.05， P<0.01）， and the levels of NLRP3， ASC proteins were decreased （P<0.01）in high dose Cranial Painkiller pills' extract powder. Meanwhile， compared with those in the model group， the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement （P<0.05， P<0.01）. ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis， and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.

ObjectiveTo explore the effects of the Tibetan medicine Sanwei Doukoutang （SWDK） on cognitive dysfunction in mice suffering from Alzheimer's disease （AD） and its related mechanism. MethodsFifty SPF 5 × FAD mice were randomly divided into model group， total ginsenoside group（0.04 g·kg^-1）， high-， medium-， and low-dose groups of SWDK （32.60， 16.30， 8.15 g·kg^-1）， with 10 mice in each group， and ten wild-type mice of the same age were used as the normal group， male and female in 1∶1. Gavage administration was performed once daily for 8 weeks. The Morris water maze test and contextual fear memory experiment were used to observe learning and memory function. Hematoxylin-eosin （HE） staining was utilized to observe the changes in the pathomorphology of brain tissue in mice. The levels of synaptophysin （SYP） and postsynaptic dense substance 95 （PSD95） in mice serum were detected by enzyme-linked immunosorbent assay （ELISA）. The positive expression of brain-derived neurotrophic factor（BDNF） in the dentate gyrus （DG） region of mouse brain tissue was observed by immunohistochemistry （IHC）. The protein levels of BDNF， Wnt family member 3A（Wnt3a）， and β-catenin were detected in the hippocampus of mice by Western blot. ResultsCompared with the normal group of mice， the model group of mice had significantly more complex swimming routes and lower swimming speed （P<0.01）， significantly lower percentage of time spent in the target quadrant （P<0.01）， and a significantly lower percentage of freezing time （P<0.05）. The number of neurons in the hippocampal region of mice was obviously reduced and unevenly arranged. The levels of SYP and PSD95（P<0.01） in the serum of mice were reduced， and the positive expression of BDNF in the DG region of the brain tissue of mice was reduced. The levels of hippocampal BDNF， Wnt3a， and β-catenin proteins in the hippocampus of mice were obviously reduced （P<0.05， P<0.01）. Compared with the model group， the mice in the SWDK group and the total ginsenoside group had significantly shorter swimming routes， the high- and medium- dose SWDK groups significantly higher swimming speeds （P<0.01）， significantly higher percentage of time spent in the target quadrant （P<0.01）， obviously higher percentage of Freezing time （P<0.05）， and obviously more neurons in the hippocampal region of the mice with tighter arrangement. The mice had elevated levels of serum SYP （P<0.05， P<0.01）， PSD95 （P<0.01）， increased BDNF-positive cells in the DG region of brain tissue， and obviously elevated levels of BDNF， Wnt3a， and β-catenin proteins in the hippocampus of mice （P<0.05， P<0.01）. ConclusionSWDK can significantly improve the cognitive dysfunction of AD mice， and its mechanism may be related to regulating the Wnt/β-catenin signaling pathway， which promotes BDNF expression and thereby enhances synaptic plasticity， allowing neuronal signaling to be restored.

ObjectiveThis paper aims to verify the therapeutic effect of Cranial Painkiller pills' extract powder prepared by the new process on the rat's trigeminal neuralgia model caused by infraorbital injection of Talci Pulvis， evaluate its potential clinical application value， and compare the therapeutic effect with that of Cranial Painkiller granules， so as to provide data support for the application of the Cranial Painkiller pills' extract powder and precise treatment. MethodsThe rat's trigeminal neuralgia model was constructed by infraorbital injection of Talci Pulvis， and the rats were randomly divided into the normal group， model group， carbamazepine group （60 mg·kg^-1）， Cranial Painkiller granules group （2.70 g·kg^-1）， and low， medium， and high dosage groups of Cranial Painkiller pills' extract powder （1.35， 2.70， 5.40 g·kg^-1） according to the basal mechanical pain thresholds， and there were 10 rats in each group. The drug was administered by gavage to each group 2 h after modeling， and distilled water was given by gavage to the normal and model groups under the same conditions once a day for 10 d. Von Frey brushes were used to measure mechanical pain thresholds in rats. Hematoxylin-eosin （HE） staining was used to detect pathological changes in the trigeminal ganglion， and enzyme-linked immunosorbent assay （ELISA） was used to detect the inflammatory factors interleukin-1 （IL-1）， interleukin-6 （IL-6）， interleukin-8 （IL-8）， and tumor necrosis factor-α （TNF-α） levels in rat serum， as well as neuropeptide substance P （SP） and β-endorphin （β-EP） levels in rat brain tissue. Western blot technique was used to detect the levels of NLRP3， ASC， Caspase-1， and OTULIN proteins in rat brain tissue. ResultsCompared with the normal group， the pain threshold of rats in the model group showed a continuous significant decrease （P<0.01）. The pathological damage of brain tissue was significant （P<0.01）， and the inflammatory levels of IL-1， IL-6， IL-8， and TNF-α in serum were significantly elevated （P<0.01）. The level of the SP in the brain tissue was significantly elevated （P<0.01）， and the level of β-EP was significantly reduced （P<0.01）， while the level of OTULIN was significantly reduced， and NLRP3， ASC， and Caspase-1 protein levels were significantly elevated （P<0.01）. After administration of the drug， compared with the model group， the pain threshold of each dose group of the Cranial Painkiller pills' extract powder and the Cranial Painkiller granules group significantly increased （P<0.01）. The inflammatory levels of IL-1， IL-6， IL-8， and TNF-α and SP levels significantly decreased （P<0.01）， and the β-EP levels were significantly elevated （P<0.01）， while the levels of OTULIN protein were significantly elevated （P<0.05， P<0.01）， and the levels of NLRP3， ASC proteins were decreased （P<0.01）in high dose Cranial Painkiller pills' extract powder. Meanwhile， compared with those in the model group， the trigeminal ganglion lesions of rats in the Cranial Painkiller pills' extract powder and Cranial Painkiller granules groups showed different degrees of improvement （P<0.05， P<0.01）. ConclusionThe Cranial Painkiller pills' extract powder has significant therapeutic effects on the rat model of trigeminal neuralgia induced by infraorbital injection of Talci Pulvis， and its mechanism is related to the improvement of OTULIN-regulated neuroinflammation.

ObjectiveTo explore the effects of the Tibetan medicine Sanwei Doukoutang （SWDK） on cognitive dysfunction in mice suffering from Alzheimer's disease （AD） and its related mechanism. MethodsFifty SPF 5 × FAD mice were randomly divided into model group， total ginsenoside group（0.04 g·kg^-1）， high-， medium-， and low-dose groups of SWDK （32.60， 16.30， 8.15 g·kg^-1）， with 10 mice in each group， and ten wild-type mice of the same age were used as the normal group， male and female in 1∶1. Gavage administration was performed once daily for 8 weeks. The Morris water maze test and contextual fear memory experiment were used to observe learning and memory function. Hematoxylin-eosin （HE） staining was utilized to observe the changes in the pathomorphology of brain tissue in mice. The levels of synaptophysin （SYP） and postsynaptic dense substance 95 （PSD95） in mice serum were detected by enzyme-linked immunosorbent assay （ELISA）. The positive expression of brain-derived neurotrophic factor（BDNF） in the dentate gyrus （DG） region of mouse brain tissue was observed by immunohistochemistry （IHC）. The protein levels of BDNF， Wnt family member 3A（Wnt3a）， and β-catenin were detected in the hippocampus of mice by Western blot. ResultsCompared with the normal group of mice， the model group of mice had significantly more complex swimming routes and lower swimming speed （P<0.01）， significantly lower percentage of time spent in the target quadrant （P<0.01）， and a significantly lower percentage of freezing time （P<0.05）. The number of neurons in the hippocampal region of mice was obviously reduced and unevenly arranged. The levels of SYP and PSD95（P<0.01） in the serum of mice were reduced， and the positive expression of BDNF in the DG region of the brain tissue of mice was reduced. The levels of hippocampal BDNF， Wnt3a， and β-catenin proteins in the hippocampus of mice were obviously reduced （P<0.05， P<0.01）. Compared with the model group， the mice in the SWDK group and the total ginsenoside group had significantly shorter swimming routes， the high- and medium- dose SWDK groups significantly higher swimming speeds （P<0.01）， significantly higher percentage of time spent in the target quadrant （P<0.01）， obviously higher percentage of Freezing time （P<0.05）， and obviously more neurons in the hippocampal region of the mice with tighter arrangement. The mice had elevated levels of serum SYP （P<0.05， P<0.01）， PSD95 （P<0.01）， increased BDNF-positive cells in the DG region of brain tissue， and obviously elevated levels of BDNF， Wnt3a， and β-catenin proteins in the hippocampus of mice （P<0.05， P<0.01）. ConclusionSWDK can significantly improve the cognitive dysfunction of AD mice， and its mechanism may be related to regulating the Wnt/β-catenin signaling pathway， which promotes BDNF expression and thereby enhances synaptic plasticity， allowing neuronal signaling to be restored.

Objective: To investigate the effects of usnic acid(UA) on proliferation, cell cycle, apoptosis and autophagy of lung cancer cells A549 and NCI-H358. Methods: The CCK-8 method was used to detect the inhibitory effect of UA on two kinds of lung cancer cells proliferation. Flow cytometry was used to detect the cell cycle arrest effect of UA on two types of lung cancer cells. The fluorescence amount of UA-induced reactive oxygen species(ROS) in two kinds of lung cancer cells were detected by DCFH-DA probe assay and flow cytometry. Western blot was used to detect the expression of apoptosis related proteins Bax, Caspase-3, Cleaved-Caspase-3, and autophagy related proteins LC3-Ⅰ and LC3-Ⅱ in two types of lung cancer cells after treatment with UA and UA+ROS inhibition. Results: (1) Usnic acid reduced the survival rates of two types of cells and had a stronger ability to inhibit the proliferation of A549 cells than NCI-H358 cells.(2) Usnic acid blocked A549 cells in the G0/G1phase, while NCI-H358 cells in the G2/M and S phases.(3) Usnic acid induced an increase in ROS content in two types of cells. Compared to A549, NCI-H358 cells showed a greater increase in ROS, and the ROS inhibitor reduced the intracellular ROS increase induced by UA.(4) Usnic acid induced high expression of apoptosis-related proteins Bax and Cleaved Caspase-3 and increased the ratio of autophagy-related proteins LC3-Ⅱ/LC3-Ⅰ in both lung adenocarcinoma cells, and its pro-apoptotic and autophagic effects were stronger in A549 than in NCI-H358. After ROS inhibition, the expression levels of Bax and Cleaved Caspase-3 and the LC3-Ⅱ/LC3-Ⅰ ratio of both lung adenocarcinoma cells decreased, and the decrease was greater in NCI-H358 cells. Conclusion Usnic acid inhibits the proliferation of lung cancer A549 and NCI-H358 cells, induces cell cycle arrest, and induces apoptosis and autophagy in cancer cells. The inhibitory and killing effect of UA on A549 cells is stronger than that on NCI-H358 cells. In this case, the induced cell ROS are involved in the action of UA in two types of lung cancer cells. In contrast to A549 cells,ROS might play a more significant role in mediating the apoptosis and autophagy induced by usnic acid in NCI-H358 cells.

AIM:To investigate the role of ClC-3 chloride channel in the promotion of radio sensitization of na-sopharyngeal carcinoma CNE-2Z cells by dihydroartemisinin(DHA).METHODS:MTT was used to detect the inhibito-ry effect of DHA on the viability of CNE-2Z cells and normal nasopharyngeal epithelial NP69-SV40T cells,the radio sensi-tization effect of DHA on CNE-2Z cells was detected by cloning assay,the expression of ClC-3 protein was detected by Western blot,the expression of ClC-3 protein was down-regulated by siRNA technology,and the chlorine current of cells was recorded by whole cell patch-clamp technology.RESULTS:(1)Compared with NP69-SV40T cells,DHA selective-ly inhibited the proliferation of CNE-2Z cells,with IC10 values of(13.020±4.831)μmol/L and(5.244±1.050)μmol/L,respectively(P＜0.01).(2)The results of clonal formation experiments showed that DHA had a radio sensitizing effect on CNE-2Z cells,with a radio sensitization ratio of 1.9.(3)DHA could activate the chlorine channel of CNE-2Z cells and produce an outward chlorine current,but had no effect on the chlorine channel of NP69-SV40T cells.(4)DHA promoted the expression of ClC-3 chloric channel protein in CNE-2Z cells(P＜0.01).(5)Chlorine channel blocker NPPB could in-hibit the radio sensitizing effect of DHA on CNE-2Z cells by 1.84 times,and also inhibited the chlorine current activated by DHA.(6)the down-regulation of CNE-2Z ClC-3 protein could inhibit the radio sensitization effect of DHA on CNE-2Z cells by 4.19 times,and the activation of chlorine current by DHA on CNE-2Z cells was no longer produced.CONCLU-SION:DHA has a radio sensitizing effect on nasopharyngeal carcinoma CNE-2Z cells,which is likely to be related to the activation of ClC-3 chloride channel.

Objective:To evaluate the value of the ROX index [blood oxygen saturation (SpO 2)/fraction of inspiration O 2 (FiO 2)/respiratory rate (RR)], ROX-heart rate (HR) index (ROX index/HR × 100), modified ROX (mROX) index [partial pressure of oxygen in the blood (PaO 2)/FiO 2/RR], and mROX-HR index (mROX index/HR × 100) in predicting prognosis for patients with acute respiratory distress syndrome (ARDS) treated with high-flow nasal cannula oxygen therapy (HFNC). Methods:The clinical data of 100 patients with ARDS who received HFNC between January 2018 and December 2022 at The Third People's Hospital of Hubei Province, Jianghan University, were retrospectively analyzed. These patients were divided into two groups based on whether HFNC treatment was successful or not: a success group with 65 patients and a failure group with 35 patients. The differences in the ROX index, ROX-HR index, mROX index, and mROX-HR index in the observation group were observed at the designated time points: 2, 12, and 24 hours after HFNC treatment. Receiver operating characteristic (ROC) curves were utilized to evaluate the value of ROX index, ROX-HR index, mROX index, and mROX-HR index in predicting the success or failure of HFNC treatment at 2, 12, and 24 hours. Cutoff values were determined.Results:There were no significant differences in age, gender, body mass index, Acute Physiology and Chronic Health Evaluation (APACHE II) score, Sequential Organ Failure Assessment score, or the proportions of underlying diseases and pulmonary causes between the success and failure groups (all P > 0.05). Furthermore, there were no significant differences in baseline HR, RR, FiO 2, SpO 2, partial pressure of carbon dioxide (PaCO 2), PaO 2, pH, lactate, oxygenation index, ROX index, mROX index, ROX-HR index, or mROX-HR index between the two groups (all P > 0.05). The ROX index in the success group at 2, 12, and 24 hours after HFNC treatment was 6.86 ± 1.09, 6.31 ± 1.61, and 8.24 ± 2.29, respectively. These values were significantly higher than those in the failure group (6.36 ± 0.67, 5.65 ± 1.44, and 5.41 ± 0.84) at the corresponding time points ( F = 5.97, 4.04, 49.40, all P < 0.05). At 2, 12, and 24 hours after HFNC treatment, the mROX index in the success group was 5.94 ± 1.28, 5.74 ± 1.23, and 8.51 ± 2.64, respectively. These values were significantly higher than those in the failure group (5.26 ± 0.74, 4.80 ± 0.97, 4.81 ± 1.17) at the corresponding time points ( F = 8.23, 15.38, 61.79, all P < 0.05). At 2, 12, and 24 hours after HFNC treatment, the ROX-HR index in the success group was 6.53 ± 1.32, 6.85 ± 1.44, and 7.57 ± 1.47, respectively. These values were significantly higher than those in the failure group (5.79 ± 1.04, 5.87 ± 1.03, 5.57 ± 0.63) at the corresponding time points ( F = 8.28, 12.61, 58.34, all P < 0.05). At 2, 12, and 24 hours after HFNC treatment, the mROX-HR index in the success group was 6.11 ± 1.30, 6.86 ± 1.13, and 7.79 ± 1.79, respectively. These values were significantly higher than those in the failure group (5.20 ± 1.06, 5.66 ± 1.46, 4.92 ± 0.90) at the corresponding time points ( F = 12.60, 20.87, 78.56, all P < 0.05). The receiver operating characteristic curve analysis revealed that the optimal thresholds were 6.56, 6.02, 6.24, and 5.25 for the ROX index, mROX index, ROX-HR index, and mROX-HR index, respectively. The area under the curve (AUC) values were 0.63, 0.66, 0.68, and 0.72, with sensitivity of 55.4%, 47.7%, 56.9%, and 76.9%, and specificity of 71.4%, 91.4%, 77.1%, and 62.9%, respectively. At 12 hours after treatment, the optimal thresholds were 6.09, 5.53, 6.52, and 5.99, with AUC values of 0.62, 0.70, 0.67, and 0.80, sensitivity of 55.4%, 53.8%, 61.5%, and 80.0%, and specificity of 74.3%, 77.1%, 71.4%, and 74.3%, respectively. At 24 hours after treatment, the optimal thresholds were 6.23, 6.4, 5.99, and 6.22, with AUC values of 0.88, 0.90, 0.91, and 0.93, sensitivity of 81.5%, 80.0%, 87.7%, and 83.1%, and specificity of 91.4%, 94.3%, 80.0%, and 91.4%, respectively. Conclusion:The use of the ROX index, mROX index, ROX-HR index, and mROX-HR index can aid in predicting the prognosis of ARDS patients. The predictive value of these indices increases as treatment time progresses. The mROX-HR index offers marked advantages during the initial stages of treatment and could serve as a reliable early predictor.

Outer membrane vesicle (OMV), originating from the outermost membrane of cells, is the extracellular vesicles released by gram-negative bacteria, containing bacterial outer membrane components such as phospholipids, lipopolysaccharide (LPS), outer membrane protein, and bacterial-specific antigens. OMV plays an important role in bacterial physiology and pathogenesis, involving in biofilm formation, horizontal gene transfer, stress and inflammatory responses, and delivery of toxins and other biomolecules. It also plays a vital role in immune regulation and the establishment and maintenance of balanced intestinal microflora. This article provides an overview of the roles of OMV in bacterial infections and immune regulation and the potential application value of OMV in tumor-targeted therapy and new vaccine preparation in the hope to provide new ideas for the prevention and treatment of bacterial infections.

Objective To discuss the clinical safety,feasibility and efficacy of transcatheter arterial infusion chemotherapy(TAI)combined with lipiodol chemoembolization in the treatment of advanced colorectal cancer(CRC).Methods The clinical data of 37 patients with advanced CRC,who received TAI combined with lipiodol chemoembolization at the First Affiliated Hospital of Zhengzhou University of China between June 2016 and December 2022,were retrospectively analyzed.The clinical efficacy was evaluated,the progression-free survival(PFS)and the serious complications were recorded.Results A total of 55 times of TAI combined with lipiodol chemoembolization procedures were successfully accomplished in the 37 patients.The mean used amount of lipiodol emulsion was 2.9 mL(0.8-10 mL).No serious complications such as bleeding and intestinal perforation occurred.The median follow-up time was 24 months(range of 3-48 months).The postoperative one-month,3-month,6-month and 12-month objective remission rates(ORR)were 67.6%(25/37),67.6%(25/37),64.9%(24/37)and 56.8%(21/37)respectively,and the postoperative one-month,3-month,6-month and 12-month disease control rates(DCR)were 91.9%(34/37),91.9%(34/37),89.2%(33/37)and 81.1%(30/37)respectively.The median PFS was 16 months(range of 2-47 months).As of the last follow-up,22 patients survived and 15 patients died of terminal stage of tumor.Conclusion Preliminary results of this study indicate that TAI combined with lipiodol chemoembolization is clinically safe and effective for advanced CRC,and it provide a new therapeutic method for patients with advanced CRC.

Objective To evaluate the clinical value of transarterial catheterization C-arm CT perfusion scanning technique during prostatic artery embolization(PAE)for benign prostatic hyperplasia(BPH).Methods The clinical data of 46 patients with BPH received PAE were analyzed retrospectively.All patients underwent prostatic artery(PA)digital subtraction angiography(DSA)and C-arm CT perfusion scanning to identify PA and prevent non-target organ embolization.The final recognization of PA was consulted by three senior doctors.After C-arm CT confirmation,PA was embolized with 100-300 μm polyvinyl alcohol(PVA)particles or microspheres under fluoroscopy.The postoperative complications and 3-month clinical efficacy were observed.Results A total of 106 vessels were angioraphed in 46 patients,with 83 PA vessels and 23 non-PA vessels.PA was identified by DSA and C-arm CT with sensitivity of 81.9%(68/83)and 100%(83/83),respectively,which showed significance(χ2=22.3,P<0.01).Non-PA was identified by DSA and C-arm CT with specificity of 73.9%(17/23)and 100%(23/23),which showed significance(χ2=9.2,P=0.02).No serious complications were observed and 3-month clincial efficacy was 91.3%.Conclusion Transarterial catheterization C-arm CT perfusion scanning technique can accurately identify PA,reduce PA leakage and prevent non-target organ embolization.