OBJECTIVE: To observe the clinical effect of acupuncture-moxibustion therapy of Daoqi Tongluo (conducting qi and unblocking collateral) on chronic primary tinnitus. METHODS: A total of 32 patients with chronic primary tinnitus were included and treated with the acupuncture-moxibustion therapy of Daoqi Tongluo. This regimen was composed of abdominal acupuncture, body acupuncture, warm needling and posterior-auricular local flashing cupping, Zhongwan (CV12), Guanyuan (CV6) and Yindu (KI9), Tinggong (SI19), Cong'er point, Waiguan (TE5) of the affected side, etc. are selected. The treatment was given once every two days, 3 treatments a week; and one course of intervention was required, with 10 treatments included. Before and after treatment, the scores of tinnitus handicap inventory (THI), tinnitus evaluation questionnaire (TEQ), self-rating scale of sleep (SRSS), self-rating anxiety scale (SAS), and self-rating depression scale (SDS) were observed, and the clinical effect was evaluated. RESULTS: After interventions, the scores of THI, TEQ, SRSS, SAS and SDS were reduced in comparison with those before interventions in the patients (P<0.001, P<0.01, P<0.05), and the total effective rate was 71.9% (23/32). CONCLUSION Acupuncture-moxibustion therapy of Daoqi Tongluo is effective on chronic primary tinnitus and this therapy can alleviate tinnitus degree, improve sleep quality and attenuate the anxious and depressive emotion of the patients.
Adult ; Aged ; Female ; Humans ; Male ; Middle Aged ; Young Adult ; Acupuncture Points ; Acupuncture Therapy ; Chronic Disease/therapy* ; Moxibustion ; Tinnitus/psychology* ; Treatment Outcome

The continuous emergence of SARS-CoV-2 variants as well as other potential future coronavirus has challenged the effectiveness of current COVID-19 vaccines. Therefore, there remains a need for alternative antivirals that target processes less susceptible to mutations, such as the formation of six-helix bundle (6-HB) during the viral fusion step of host cell entry. In this study, a novel high-throughput screening (HTS) assay employing a yeast-two-hybrid (Y2H) system was established to identify inhibitors of HR1/HR2 interaction. The compound IMB-9C, which achieved single-digit micromolar inhibition of SARS-CoV-2 and its Omicron variants with low cytotoxicity, was selected. IMB-9C effectively blocks the HR1/HR2 interaction in vitro and inhibits SARS-CoV-2-S-mediated cell-cell fusion. It binds to both HR1 and HR2 through non-covalent interaction and influences the secondary structure of HR1/HR2 complex. In addition, virtual docking and site-mutagenesis results suggest that amino acid residues A930, I931, K933, T941, and L945 are critical for IMB-9C binding to HR1. Collectively, in this study, we have developed a novel screening method for HR1/HR2 interaction inhibitors and identified IMB-9C as a potential antiviral small molecule against COVID-19 and its variants.

Objective To investigate the mechanism of tea polyphenols(TP)for regulating NLRP3 inflammasomes and alleviating acute lung injury in septic mice.Methods Sixty C57BL/6 mice were randomly assigned into sham-operated,cecal ligation and puncture(CLP)and CLP+TP treatment groups,and survival of the mice was recorded after modeling in each group.The lung wet/dry weight ratio and myeloperoxidase(MPO)activity were determined,and lung injury of the mice was evaluated using HE staining and acute lung injury score.The expressions of IL-1β,TNF-α,IL-6,NLRP3,caspase-1 p10,ASC,MPO,and caspase-8 in the lung tissue were detected using ELISA,Western blotting,or immunohistochemical staining.MDA and H2O2 levels in the lungs were detected to evaluate the level of oxidative stress.Immunofluorescence assay was used to investigate the co-localization of NLRP3 and NOX4.Results The postoperative mortality rate at 72 h,lung wet/dry weight ratio,MPO level and acute lung injury scores were significantly lower in CLP+TP group than in CLP group(P<0.05).Treatment with TP significantly reduced the expressions of NLRP3-related inflammatory factors(P<0.05)and lowered MDA and H2O2 levels in the lung tissue of the septic mice(P<0.05).Immunofluorescence co-staining showed a lower level of NOX4 and NLRP3 co-localization in CLP+TP group than in CLP group.Conclusion TP inhibits NLRP3 inflammasome-associated inflammation to alleviate CLP-induced acute lung injury in mice through a regulatory mechanism that inhibits NOX4 expression and reduces oxidative stress in the lung tissue.

Objective To investigate the effect of glycine N-methyl transferase (GNMT)on intrauterine adhesion (IUA)fibrosis and its related mechanism.Methods In vivo experiment:A total of 36 healthy female SD rats (SPF grade,6～8 weeks old and weighing from 180～220 g)were subjected in this study.IUA model of SD rats and IUA model of GNMT overexpressed rats were established.RT-qPCR and immunofluorescence assay were applied to detect GNMT expression level in normal uterus and model group.RT-qPCR and Western blotting were used to detect the mRNA and protein levels of fibrosis-related molecules and the activation of TGF-β1/Smad3 signaling pathway in each group.The number of endometrial glands in each group was observed by HE staining.Masson staining was used to analyze the severity of endometrial fibrosis in each group.In vitro experiment:transformed human endometrial stromal cells (THESCs)fibrotic phenotype model was constructed using TGF-β1,and THESCs stably transfected with GNMT overexpression lentvirus were treated with TGF-β1.RT-qPCR and Western blotting were used to detect the mRNA and protein expression of fibrosis-related molecules.The expression of TGF-β1/Smad3 signaling pathway was detected by Western blotting.TGF-β1/Smad3 signaling pathway was activated by TGF-β1/Smad signaling pathway activator (SRI-011381),and the expression of TGF-β1/Smad3 signaling pathway and key molecular proteins of fibrosis phenotype was measured with Western blotting.Results In vivo experiment,the mRNA and protein expression levels of GNMT were significantly decreased in the IUA rats than the control rats (P<0.05).Overexpression of GNMT decreased the mRNA and protein levels of fibrosis related molecules,Collagen Ⅰ,Collagen Ⅲ and FN in the IUA rats (P<0.05),and decreased the phosphorylation levels of TGF-β1 and its downstream Smad3 protein (P<0.05).HE and Masson staining showed that overexpression of GNMT could increase the number of endometrial glands and reduce the severity of fibrosis in the IUA rats (P<0.05).In vitro experiments:overexpression of GNMT decreased the mRNA and protein levels of Collagen Ⅰ,Collagen Ⅲ and FN associated with fibrotic phenotype of THESCs (P<0.05),and reduced the phosphorylation level of Smad3 protein,downstream of TGF-β1 (P<0.05).After activation of TGF-β1/Smad3 signaling pathway,the protein levels of TGF-β1/Smad3 signaling pathway and downstream fibrosis phenotype molecules,Collagen Ⅲ and FN,were significantly decreased in the LV-GNMT+SRI-011381 group.Conclusion Overexpression of GNMT can inhibit endometrial fibrosis by regulating TGF-β1/Smad3 signaling pathway,thus achieving therapeutic effect on IUA.

Objective To investigate the mechanism of tea polyphenols(TP)for regulating NLRP3 inflammasomes and alleviating acute lung injury in septic mice.Methods Sixty C57BL/6 mice were randomly assigned into sham-operated,cecal ligation and puncture(CLP)and CLP+TP treatment groups,and survival of the mice was recorded after modeling in each group.The lung wet/dry weight ratio and myeloperoxidase(MPO)activity were determined,and lung injury of the mice was evaluated using HE staining and acute lung injury score.The expressions of IL-1β,TNF-α,IL-6,NLRP3,caspase-1 p10,ASC,MPO,and caspase-8 in the lung tissue were detected using ELISA,Western blotting,or immunohistochemical staining.MDA and H2O2 levels in the lungs were detected to evaluate the level of oxidative stress.Immunofluorescence assay was used to investigate the co-localization of NLRP3 and NOX4.Results The postoperative mortality rate at 72 h,lung wet/dry weight ratio,MPO level and acute lung injury scores were significantly lower in CLP+TP group than in CLP group(P<0.05).Treatment with TP significantly reduced the expressions of NLRP3-related inflammatory factors(P<0.05)and lowered MDA and H2O2 levels in the lung tissue of the septic mice(P<0.05).Immunofluorescence co-staining showed a lower level of NOX4 and NLRP3 co-localization in CLP+TP group than in CLP group.Conclusion TP inhibits NLRP3 inflammasome-associated inflammation to alleviate CLP-induced acute lung injury in mice through a regulatory mechanism that inhibits NOX4 expression and reduces oxidative stress in the lung tissue.

ObjectiveTo establish a quantitative analysis multi-components by single marker method （QAMS） for five main components （aucubin， geniposidic acid， chlorogenic acid， asperuloside and rutin） in Eucommiae Folium， to verify its feasibility and applicability in the determination of Eucommiae Folium， so as to provide a scientific basis for the development of quality standard of this herb. MethodHigh performance liquid chromatography was performed on a Welch Boltmatetm™ C₁₈ column （4.6 mm×100 mm， 2.7 μm） with methanol （A）-0.2% phosphoric acid aqueous solution （B） as the mobile phase for gradient elution （0-8 min， 3%A； 8-10 min， 3%-11%A； 10-26 min， 11%A； 26-27 min， 11%-25%A； 27-60 min， 25%-32%A）， the column temperature was set at 30 ℃， the flow rate was 0.6 mL·min^-1， the detection wavelengths were at 210 nm and 254 nm. Chlorogenic acid was used as an internal reference to establish the relative correction factors （f） between it and the other four components， and the contents of the five components in 14 batches of Eucommiae Folium were determined by QAMS and external standard method （ESM）， respectively. ResultThe f values of chlorogenic acid to aucubin， geniposidic acid， asperuloside and rutin were 3.13， 1.45， 2.64 and 0.56， respectively. Repeatability was good under different experimental conditions， relative standard deviation （RSD） was <5.0%. The contents of aucubin， geniposidic acid， chlorogenic acid， asperuloside and rutin in 14 batches of Eucommiae Folium were 1.340-28.975， 0.252-36.086， 10.016-27.443， 1.396-8.646， 0.533-1.766 mg·g^-1， respectively. There were no significant difference between content results of QAMS and that of ESM （RSD<5.0%）. ConclusionQAMS established with chlorogenic acid as the internal reference can be used to determine the contents of five components in Eucommiae Folium， and this method is simple and accurate. After comprehensive evaluation， the quality standard of Eucommiae Folium in subsequent editions of Chinese Pharmacopoeia is suggested that three main active components， chlorogenic acid， aucubin and geniposidic acid， are selected as quality markers， and their content limits are recommended not less than 1.5%， 1.0% and 1.0%， respectively. This quality standard draft can avoid the potential quality risk due to poor specificity and low content limit of the index component （chlorogenic acid） in the previous editions of Chinese Pharmacopoeia.

Resection of oral and maxillofacial tumors is often accompanied by the inferior alveolar nerve neurectomy, resulting in abnormal sensation in lower lip. It is generally believed that spontaneous sensory recovery in this nerve injury is difficult. However, during our follow-up, patients with inferior alveolar nerve sacrifice showed different degrees of lower lip sensory recovery. In this study, a prospective cohort study was conducted to demonstrate this phenomenon and analyze the factors influencing sensory recovery. A mental nerve transection model of Thy1-YFP mice and tissue clearing technique were used to explore possible mechanisms in this process. Gene silencing and overexpression experiments were then conducted to detect the changes in cell morphology and molecular markers. In our follow-up, 75% of patients with unilateral inferior alveolar nerve neurectomy had complete sensory recovery of the lower lip 12 months postoperatively. Patients with younger age, malignant tumors, and preservation of ipsilateral buccal and lingual nerves had a shorter recovery time. The buccal nerve collateral sprouting compensation was observed in the lower lip tissue of Thy1-YFP mice. ApoD was demonstrated to be involved in axon growth and peripheral nerve sensory recovery in the animal model. TGF-β inhibited the expression of STAT3 and the transcription of ApoD in Schwann cells through Zfp423. Overall, after sacrificing the inferior alveolar nerve, the collateral compensation of the ipsilateral buccal nerve could innervate the sensation. And this process was regulated by TGF-β-Zfp423-ApoD pathway.
Mice ; Animals ; Lip/innervation* ; Prospective Studies ; Mandibular Nerve/pathology* ; Sensation/physiology* ; Trigeminal Nerve Injuries/pathology*

Objective:To investigate the method and effect of correcting upper eyelid depression with released orbital diaphragmatic fat.Methods:From January 2017 to June 2020, there were 26 female patients with upper eyelid depression in this department of Hebei Chengde Stomatological Hospital. The patients aged 22-54 years, with average 43 years. An external mass orbital diaphragm fat release reverse folding repositioning with fat placement in the upper lid depression was performed in 26 patients. An incision about 4 mm long was made in the middle and lateral of the orbital diaphragm to ensure the integrity of the orbital diaphragm as far as possible. The photos of beauty seekers before and 6 months after operation were compared to observe the correction effect of upper eyelid depression and the shape of upper eyelid.Results:All 26 patients had one-stage healing of the surgical incision without infection, with inconspicuous incisional scarring. At 6 months postoperatively, 18 patients had satisfactory results; 1 patient had improved results due to insufficient fat; and 1 patient had poor results due to too little fat and poor filler placement, which was later improved by autologous fat grafting.Conclusions:External mass orbital fat release reverse folding repositioning for correction of upper eyelid depressions is a surgical procedure with obvious results, little trauma, and following the principle of anatomical recovery, which is worthy of clinical application.

Objective:To investigate the relationship between electrocardiographic (ECG) changes and prognosis of paraquat poisoning patients, so as to provide evidence for the condition assessment in paraquat poisoning patients.Methods:In January 2022, The clinical data of paraquat poisoning patients were retrospectively analyzed in the First Affiliated Hospital of Wenzhou Medical University from January 1, 2016 to December 31, 2021. The patients' basic information (age, sex, underlying disease, and occupation) and the ECG within 24 hours were collected, and the data were statistically analyzed by SPSS 22.0. One variable analysis and multivariable logistic regression analysis were used to analyze the risk factors related to prognosis determine in their ECG. The receiver operating characteristic (ROC) curve were used to evaluate the diagnostic value of ECG indexes in patients with paraquat poisoning.Results:A total of 145 patients with paraquat poisoning were finally enrolled in this study, there were 84 patients survived and 60 patients died. One variable analysis revealed that heart rate ( P=0.000) , QTc changes ( P=0.000) , and ST-T changes ( P=0.007) of ECG had statistically significant differences in the prognosis of paraquat poisoning patients between the survival group and the death group. Multifactorial logistic results showed that heart rate ( OR=1.059, 95% CI: 1.033~1.086) and QTc ( OR=1.015, 95% CI: 1.000~1.029) were independent risk factors for death diagnosis of patients with paraquat poisoning ( P<0.05) . ROC curve analysis revealed that the area under the cure (AUC) of the prediction model constructed based on heart rate and QTc was 0.832 (95% CI: 0.765~0.899) , with the best diagnostic efficacy. Conclusion:Heart rate, QTc and the prediction model constructed based on both can be used as prognostic indicators for the diagnosis of patients with paraquat poisoning, and which have reference value for clinical prognosis diagnosis.

Objective:To investigate the relationship between electrocardiographic (ECG) changes and prognosis of paraquat poisoning patients, so as to provide evidence for the condition assessment in paraquat poisoning patients.Methods:In January 2022, The clinical data of paraquat poisoning patients were retrospectively analyzed in the First Affiliated Hospital of Wenzhou Medical University from January 1, 2016 to December 31, 2021. The patients' basic information (age, sex, underlying disease, and occupation) and the ECG within 24 hours were collected, and the data were statistically analyzed by SPSS 22.0. One variable analysis and multivariable logistic regression analysis were used to analyze the risk factors related to prognosis determine in their ECG. The receiver operating characteristic (ROC) curve were used to evaluate the diagnostic value of ECG indexes in patients with paraquat poisoning.Results:A total of 145 patients with paraquat poisoning were finally enrolled in this study, there were 84 patients survived and 60 patients died. One variable analysis revealed that heart rate ( P=0.000) , QTc changes ( P=0.000) , and ST-T changes ( P=0.007) of ECG had statistically significant differences in the prognosis of paraquat poisoning patients between the survival group and the death group. Multifactorial logistic results showed that heart rate ( OR=1.059, 95% CI: 1.033~1.086) and QTc ( OR=1.015, 95% CI: 1.000~1.029) were independent risk factors for death diagnosis of patients with paraquat poisoning ( P<0.05) . ROC curve analysis revealed that the area under the cure (AUC) of the prediction model constructed based on heart rate and QTc was 0.832 (95% CI: 0.765~0.899) , with the best diagnostic efficacy. Conclusion:Heart rate, QTc and the prediction model constructed based on both can be used as prognostic indicators for the diagnosis of patients with paraquat poisoning, and which have reference value for clinical prognosis diagnosis.