Objective:To explore mechanism of Fasudil reducing Aβ1-42 induced BV2 cell injury based on NLRP3 inflamma-some.Methods:BV2 cells were divided into:normal control group,Aβ stimulation group,Aβ+Fasudil intervention group,Aβ+MCC950(NLRP3 inhibitor)intervention group.Cell morphology was observed under microscope.Cell activity was determined of by CCK8.NO release was measured by Griess.NLRP3,caspase 1 and IL-18 expressions were detected by immunofluorescence staining.NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were detected by Western blot.Results:Compared with normal control group,BV2 cells in Aβ stimulation group were activated and showed amoeba-like shape,cell activity was decreased,NO production was increased,NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were increased.Fasudil intervention and MCC950 intervention inhibited cell injury induced by Aβ1-42 in which BV2 cell morphology tended to be normal,cell activity was increased,while produc-tion of NO was reduced,and NLRP3,ASC,caspase 1,IL-1β and IL-18 expressions were down-regulated,there was no significant difference between Fasudil intervention group and MCC950 intervention group.Conclusion:Fasudil may alleviate Aβ1-42 induced BV2 cell injury and inflammatory reaction by inhibiting NLRP3 inflammasome activation.

Objective To predict the potential geographic distribution of Oncomelania hupensis in Yunnan Province using random forest (RF) and maximum entropy (MaxEnt) models, so as to provide insights into O. hupensis surveillance and control in Yunnan Province. Methods The O. hupensis snail survey data in Yunnan Province from 2015 to 2016 were collected and converted into O. hupensis snail distribution site data. Data of 22 environmental variables in Yunnan Province were collected, including twelve climate variables (annual potential evapotranspiration, annual mean ground surface temperature, annual precipitation, annual mean air pressure, annual mean relative humidity, annual sunshine duration, annual mean air temperature, annual mean wind speed, ≥ 0 ℃ annual accumulated temperature, ≥ 10 ℃ annual accumulated temperature, aridity and index of moisture), eight geographical variables (normalized difference vegetation index, landform type, land use type, altitude, soil type, soil textureclay content, soil texture-sand content and soil texture-silt content) and two population and economic variables (gross domestic product and population). Variables were screened with Pearson correlation test and variance inflation factor (VIF) test. The RF and MaxEnt models and the ensemble model were created using the biomod2 package of the software R 4.2.1, and the potential distribution of O. hupensis snails after 2016 was predicted in Yunnan Province. The predictive effects of models were evaluated through cross-validation and independent tests, and the area under the receiver operating characteristic curve (AUC), true skill statistics (TSS) and Kappa statistics were used for model evaluation. In addition, the importance of environmental variables was analyzed, the contribution of environmental variables output by the models with AUC values of > 0.950 and TSS values of > 0.850 were selected for normalization processing, and the importance percentage of environmental variables was obtained to analyze the importance of environmental variables. Results Data of 148 O. hupensis snail distribution sites and 15 environmental variables were included in training sets of RF and MaxEnt models, and both RF and MaxEnt models had high predictive performance, with both mean AUC values of > 0.900 and all mean TSS values and Kappa values of > 0.800, and significant differences in the AUC (t = 19.862, P < 0.05), TSS (t = 10.140, P < 0.05) and Kappa values (t = 10.237, P < 0.05) between two models. The AUC, TSS and Kappa values of the ensemble model were 0.996, 0.954 and 0.920, respectively. Independent data verification showed that the AUC, TSS and Kappa values of the RF model and the ensemble model were all 1, which still showed high performance in unknown data modeling, and the MaxEnt model showed poor performance, with TSS and Kappa values of 0 for 24%(24/100) of the modeling results. The modeling results of 79 RF models, 38 MaxEnt models and their ensemble models with AUC values of > 0.950 and TSS values of > 0.850 were included in the evaluation of importance of environmental variables. The importance of annual sunshine duration (SSD) was 32.989%, 37.847% and 46.315% in the RF model, the MaxEnt model and their ensemble model, while the importance of annual mean relative humidity (RHU) was 30.947%, 15.921% and 28.121%, respectively. Important environment variables were concentrated in modeling results of the RF model, dispersed in modeling results of the MaxEnt model, and most concentrated in modeling results of the ensemble model. The potential distribution of O. hupensis snails after 2016 was predicted to be relatively concentrated in Yunnan Province by the RF model and relatively large by the MaxEnt model, and the distribution of O. hupensis snails predicted by the ensemble model was mostly the joint distribution of O. hupensis snails predicted by RF and MaxEnt models. Conclusions Both RF and MaxEnt models are effective to predict the potential distribution of O. hupensis snails in Yunnan Province, which facilitates targeted O. hupensis snail control.

The study was designed to investigate the peripheral anti-inflammatory effect of nicotinamide riboside(NR)in experimental autoimmune encephalomyelitis(EAE)mice and its mechanisms.Female C57BL/6 mice were induced by MOG35-55 to prepare EAE model,which then randomly divided into EAE model group and NR treatment group.Mice in EAE model group were given normal saline at a dose of 200 μl/d and mice in NR treatment group were given NR at a dose of 500 mg/kg(200 μl/d)by intragastric administration.Clinical score and body weight of mice in each group were observed and recorded.After mice were sacrificed on the 28th day after immunization,frozen sections of spleen and spinal cord were prepared and proteins of spinal cord were extracted.HE staining was used to detect peripheral inflammatory cells infiltrating spinal cord;immunofluorescence staining was used to detect the number of CD4+T cells and CD68+macrophages in spinal cord of mice;Western blot was used to detect the expression of IFN-γ and IL-1β in spinal cord of mice;immunofluorescence staining was used to detect the number of ROCK-Ⅰ+cells,TLR4+cells,p-NF-κB+cells,TNF-α+cells,IL-1β+cells,IFN-γ+cells,IL-6+cells,IL-10+cells,IL-17+cells,iNOS+cells and Arg-1+cells in spleen of mice.Data showed that compared with EAE model group,NR significantly delayed the onset time of EAE mice(P<0.05),decreased clinical score(P<0.05 or P<0.01),alleviated weight loss,prevented peripheral inflammatory cells from infiltrating spinal cord,decreased the number of CD4+T cells and CD68+macrophages in spinal cord(P<0.01),down-regulated the expression of IFN-γ and IL-1β of spinal cord(P<0.05),inhibited the expression of ROCK-Ⅰ,TLR4 and p-NF-κB in spleen of mice(P<0.01),reduced the secretion of IFN-γ,iNOS,IL-6 and other pro-inflammatory factors in spleen(P<0.05 or P<0.01),and increased the secretion of anti-inflammatory factors Arg-1 and IL-10 in spleen(P<0.05 or P<0.01).In conclusion,NR can effectively alleviate the clinical symptoms of EAE mice and significantly reduce inflammatory response of peripheral and central nervous system,and its mechanism may be related to the inhibition of Rho/ROCK signaling pathway and TLR4/NF-κB signaling pathway in spleen of EAE mice.

Objective To explore the effect of knocking down Rho-associated coiled-coil kinase (ROCK2) gene on the cognitive function of amyloid precursor protein/presenilin-1 (APP/PS1) double transgenic mice and its mechanism. Methods APP/PS1 double transgenic mice were randomly divided into AD model group (AD group), ROCK2 gene knock-down group (shROCK2 group), ROCK2 gene knock-down control group (shNCgroup), and wild-type C57BL/6 mice of the same age served as the wild-type control (WT group). Morris water maze and Y maze were employed to test the cognitive function of mice. Neuron morphology was detected by Nissl staining. Immunofluorescence histochemical staining was used to detect the expression of phosphorylated dynamin-related protein 1 (p-Drp1) and mitochondrial fusion 1 (Mfn1). Western blot analysis was used to detect the expression ROCK2, cleaved-caspase-3 (c-caspase-3), B-cell lymphoma 2 (Bcl2), Bcl2-related protein X (BAX), p-Drp1, mitochondrial fission 1 (Fis1), optic atrophy 1 (OPA1), Mfn1 and Mfn2. Results Compared with AD group mice, the expression of ROCK2 in shROCK2 group mice was significantly reduced; the cognitive function was significantly improved with the number of neurons in the hippocampal CA3 and DG areas increasing, and nissl bodies were deeply stained; the expression of c-caspase-3 and BAX was decreased, while the expression of Bcl2 was increased; the expression of mitochondrial division related proteins p-Drp1 and Fis1 were decreased, while the expression of mitochondrial fusion-related proteins OPA1, Mfn1 and Mfn2 were increased. Conclusion Knock-down of ROCK2 gene can significantly improve the cognitive function and inhibit the apoptosis of nerve cells of APP/PS1 mice. The mechanism may be related to promoting mitochondrial fusion and inhibiting its division.
Animals ; Mice ; Alzheimer Disease/pathology* ; Amyloid beta-Peptides/metabolism* ; Amyloid beta-Protein Precursor ; Apoptosis/genetics* ; bcl-2-Associated X Protein ; Caspase 3 ; Cognition ; Disease Models, Animal ; Mice, Inbred C57BL ; Mice, Transgenic ; Mitochondrial Dynamics/genetics*

OBJECTIVE To establish the correlation evaluation and quality evaluation method of HPLC fingerprint grade of color sorting Lonicerae Japonicae Flos, and provide technical basis for the grade standard of color sorting Lonicerae Japonicae Flos. METHODS The chromatographic column was SVEA C18(250 mm×4.6 mm, 5 μm); mobile phase was acetonitrile(A)- 0.2% formic acid aqueous solution(B); gradient elution; injection volume was 10 μL; detection wave length was 245 nm; volume flow rate was 0.5 mL·min-1; column temperature was 38 ℃. The common peak determination and similarity evaluation of HPLC chromatogram data were carried out by using the Similarity Evaluation System of Traditional Chinese Medicine Chromatographic Fingerprints(Version 2012); the color sorting grade evaluation was carried out by CA, PCA and PLS-DA. The first part of Chinese Pharmacopoeia 2020 Edition was used to measure the quality control indicators, and the data were analyzed comprehensively. RESULTS A total of 28 common peaks were identified in the fingerprints, and 7 components were identified. The similarity of 24 batches of color sorting grade samples was 0.936-0.968. CA and PCA divided 28 batches of Lonicerae Japonicae Flos samples into 4 categories, which were basically consistent with the classification of color sorting, and PLS-DA achieved a discrimination result that was very consistent with the classification of color sorting. The color sorting grade was negatively correlated with the diameter, flowering rate, damage rate, and luteolin content of Lonicerae Japonicae Flos. The color sorting grade was positively correlated with chlorogenic acid, 3,5-di-O-caffeoyl quinic acid and 4,5-di-O-caffeoyl quinic acid. There was a clear correlation between the color sorting of Lonicerae Japonicae Flos and established fingerprint overall. There were differences in the quality of Lonicerae Japonicae Flos in the color sorting grade. Based on the sensory indicators of diameter, flowering rate, and damage rate, the content, diameter, flowering rate, and damage rate of luteolin showed a trend from high to low, ranging from third grade>second grade>first grade>special grade. The content of three phenolic acids showed a trend from high to low, ranging from special grade>first grade>second grade>third grade. Among the special grade, the content of three phenolic acids was the highest. CONCLUSION Combining the content of luteolin and phenolic acids as evaluation and control indicators for color selection grade is feasible and scientific, which can achieve intelligent color sorting grade production of Lonicerae Japonicae Flos grade.

Dermatofibrosarcoma protuberans ( DFSP ) is very rare in clinic, and its diagnosis is not specific. According to the literature domestic and abroad, only 18 cases of patients during gestation period suffered from DFSP have been reported. This paper reported a case from the Department of Plastic Surgery, the First Affiliated Hospital of Bengbu Medical College in April 2022 diagnosed with DFSP during pregnancy.The DFSP showed rapid growth and pain during pregnancy, and stopped rapid growth after delivery. Extensive local resection was performed and the incision margin exceeded 3 cm diameter of the tumor. One-stage skin graft was used to repair the defect. HE staining showed a single spindle cell, arranged in a vortex, with large and dark nuclei. Some nuclei were heteromorphic, and few mitotic figures were observed. Postoperative pathology showed negative margins and basement. Immunohistochemistry showed CD34 (+ + ), Ki-67 (+, 10% ), bcl-2 (-), s-100 (-), SMA (-), CD163 (-), CD68 (-), which were consistent with the diagnosis of DFSP. The pathogenesis of DFSP is not yet clear. As a special population, patients during gestation period need special treatment and diagnosis. Through literature review, this paper summarizes the reports domestic and abroad, and discusses the diagnosis, treatment and prognosis of patients with DFSP during pregnancy.

Dermatofibrosarcoma protuberans ( DFSP ) is very rare in clinic, and its diagnosis is not specific. According to the literature domestic and abroad, only 18 cases of patients during gestation period suffered from DFSP have been reported. This paper reported a case from the Department of Plastic Surgery, the First Affiliated Hospital of Bengbu Medical College in April 2022 diagnosed with DFSP during pregnancy.The DFSP showed rapid growth and pain during pregnancy, and stopped rapid growth after delivery. Extensive local resection was performed and the incision margin exceeded 3 cm diameter of the tumor. One-stage skin graft was used to repair the defect. HE staining showed a single spindle cell, arranged in a vortex, with large and dark nuclei. Some nuclei were heteromorphic, and few mitotic figures were observed. Postoperative pathology showed negative margins and basement. Immunohistochemistry showed CD34 (+ + ), Ki-67 (+, 10% ), bcl-2 (-), s-100 (-), SMA (-), CD163 (-), CD68 (-), which were consistent with the diagnosis of DFSP. The pathogenesis of DFSP is not yet clear. As a special population, patients during gestation period need special treatment and diagnosis. Through literature review, this paper summarizes the reports domestic and abroad, and discusses the diagnosis, treatment and prognosis of patients with DFSP during pregnancy.

Objective:To investigate the effects of ultrasound-guided lumbar erector spinae plane block (L-ESPB) combined with general anesthesia using a laryngeal mask airway on postoperative delirium and early outcome in older adult patients undergoing hip arthroplasty.Methods:Sixty older adult patients who underwent unilateral total hip arthroplasty under general anesthesia using a laryngeal mask airway from August 2020 to May 2021 were included in this study. They were randomly assigned to observation and control groups, with 30 patients in each group. All patients underwent general anesthesia using a laryngeal mask airway. In the observation group, L-ESPB with 30 mL 0.375% ropivacaine was performed before induction of general anesthesia. Patient-controlled intravenous analgesia was performed in all patients after surgery. The Visual Analogue Scale score was > 4 points, and dezocine was intravenously injected for rescue analgesia. The amount of general anesthetics, the rate of use of vasoactive drugs, the time to pull out the laryngeal mask airway, length of stay in postanesthesia care unit, and the incidence of restlessness during the recovery period were recorded. Resting-state VAS score and Bruggrmann comfort scale score at 4, 8, 12, 24 and 48 hours after surgery, effective times of pressing the analgesia pump, the rate of rescue analgesia, the incidence of anesthesia-related adverse reactions at 48 hours after surgery, Pittsburgh Sleep Quality Index (PSQI) score at 1 day before and 1, 3 and 5 days after surgery, the incidence of postoperative delirium within 5 days after surgery, the time to get out of bed the first time, and the incidence of pulmonary infection were recorded.Results:The amount of remifentanil and propofol used during surgery were significantly lower in the observation group than those in the control group ( t = -6.80, -5.23, both P < 0.05). The rate of use of urapidil and esmolol were significantly lower in the observation group than those in the control group ( χ2 = 4.32, 5.46, both P < 0.05). The time to pull out the laryngeal mask airway, length of stay in postanesthesia care unit, and the time to get out of bed the first time in the observation group were (14 ± 2) minutes, (21 ± 2) minutes and (2.4 ± 0.5) days respectively, which were significantly shorter than those in the control group [(18 ± 2) minutes, (26 ± 3) minutes and (2.8 ± 0.7) days, t = -6.64, -7.18, -2.51, all P < 0.05]. The incidence of restlessness during the recovery period, postoperative delirium, and pulmonary infection in the observation group were 0.0%, 3.3% and 0.0% respectively, which were significantly lower than those in the control group [20.0%, 26.7%, 20.0%, all P < 0.05]. Resting-state VAS score at each time point was significantly lower in the observation group than that in the control group ( t = 3.32, 2.97, 4.33, 3.81, 3.10, all P < 0.05). Bruggrmann comfort scale score at each time point was significantly higher in the observation group than that in the control group ( t = 9.20, 8.62, 5.73, 5.72, 6.28, all P < 0.05). The effective times of pressing the analgesia pump, the rate of rescue analgesia, and the incidence of nausea and vomiting at 48 hours after surgery in the observation group were 0 (0, 0), 10.0% and 6.7% respectively, which were significantly lower than those in the control group [1(0, 2), 33.3% and 30.0%, Z = -3.41, χ2 = 4.81, 5.46, all P < 0.05]. PSQI score measured at 1, 3, and 5 days after surgery were significantly increased compared with those measured 1 day before surgery in each group ( F = 116.80, 325.64, both P < 0.001). PSQI measured at 1, 3 and 5 days after surgery was significantly lower than those in the control group ( t = -7.26, -6.41, -11.17, all P < 0.001). There were no significant differences in the incidence of dizziness, drowsiness, urinary retention and atelectasis between the two groups ( P > 0.05). Conclusion:Ultrasound-guided L-ESPB combined with general anesthesia using a laryngeal mask airway for hip arthroplasty in older adult patients can decrease the incidences of postoperative delirium and complications, which are conducive to early postoperative outcomes.

【Objective】 To construct the Rb luciferase reporter gene assay system and detect the activation ability of Rb gene for screening the targeted drugs. 【Methods】 The synthetic Rb gene sequence was annealed to form a double-stranded DNA structure and then inserted into the polyclonal site of pGL6-TA. The junction product was transformed into E.coli DH5α competent cells for expanded culture, and the constructed pGL6-Rb-Luc plasmid and pGL6-TA plasmid were transfected into HEK293 cells. The monoclonal cell line HEK293-Rb-Luc with stable expression was screened by G418, and the activation and inhibition of Rb in HEK293-Rb-Luc were tested by serum and CDK4/6 inhibitor Palbociclib. 【Results】 The sequence of Rb reaction elements in pGL6-Rb-Luc was completely correct. The recovery of serum culture significantly increased the luciferase activity in HEK293-Rb-Luc (P<0.001). Compared with 0 nmol/L, 25, 50, 75 and 100 nmol/L, CDK4/6 inhibitor Palbociclib made the inhibition rate of Rb activity rise to 6.90%, 40.23%, 50.57% and 52.07%, respectively (P<0.05). 【Conclusion】 The Rb luciferase reporter gene detection system HEK293-Rb-Luc was successfully constructed, which can effectively detect the activation level of Rb.

【Objective】 To clone the full-length of human kidney and brain protein (KIBRA) coding sequence in eukaryotic expression vector and provide a model for studying the biological function of KIBRA in breast cancer cells. 【Methods】 Total RNA of human breast cancer cell line MCF7 was extracted. After reverse transcription, the full length of KIBRA (NM_001161661.2) coding region was amplified by PCR, and cloned into eukaryotic expression vector pCMV-Blank. After identification, it was defined officially as pCMV-KIBRA. Then it was transfected into MCF7 cells, and the expression of KIBRA was detected by real-time PCR and Western blotting after 48 hours. The primary, secondary and tertiary structures and post-transcriptional modification sites of KIBRA were analyzed with bioinformatics software. 【Results】 Bacterial PCR, double enzyme digestion and DNA sequencing results showed that the correct sequence of KIBRA was inserted into the vector pCMV-KIBRA. The mRNA and protein expressions of KIBRA were significantly increased in MCF7 cells transfected with pCMV-KIBRA. Bioinformatics analysis showed that KIBRA was composed of 1119 amino acids. There were 52 phosphorylation sites, 1 acetylation site and 5 ubiquitination sites, and the protein structure was mainly α-helix and random coil. 【Conclusion】 The eukaryotic expression vector of full-length of human KIBRA coding sequence was successfully constructed and overexpressed in breast cancer cell line MCF7, which can lay a foundation for studying the biological function of KIBRA in breast cancer.