Objective:To explore whether the food additive sodium benzoate(NAB) induces pancreatic inflammation and β cell apoptosis through the benzoylation(Kbz) modification pathway.Methods:In vivo experiments: C57BL/6J male mice(8 weeks old, 18-20 g) were randomly divided into normal control group(double distilled water feeding) and NAB feeding group(1 g/kg NAB feeding). Blood glucose were measured. After 20 weeks, fasting serum insulin, interleukin(IL)-18, IL-1β, and benzoyl-CoA levels were detected by ELISA method. Bax, IL-18, Pan-Kbz and Pan-Kac were detected by immunohistochemistry staining. In vitro experiments: β-TC-6 cells were cultured with NAB(6 mmol/L) or benzoyl-CoA(100 μmol/L) as stimulator and acyltransferase P300 inhibitor A485(10 μmol/L) as intervention factor. 24 hours later, inflammation, apoptosis, insulin secretion and Pan-Kbz level were detected by qRT-PCR, ELISA and Western blotting.Results:In the in vivo experiments, compared to the NC group, mice in the NAB group exhibited impaired glucose tolerance, decreased fasting insulin levels, significantly increased serum benzoyl coenzyme A concentrations, relatively elevated pancreatic IL-1β, IL-18, and Bax protein expressions, increased levels of Pan-Kbz, while Pan-Kac levels were downregulated(all P<0.05); In vitro experiments, NAB dose-dependently inhibited insulin secretion, promoted the release of Pan-Kbz and inflammatory factors IL-18 and TNF- α, inhibited Bcl-2 expression and up-regulated Bax expression, A485 reversed NAB-induced Pan-Kbz modification, improved NAB-induced inflammation and apoptosis, and promoted insulin secretion(all P<0.05). Conclusion:NAB may induce pancreatic inflammation, β-cell apoptosis, and impair insulin secretion through Kbz modification pathway.

Objective To explore the value of multiparameter MRI for follow-up observation on changes of spinal cord microstructures in patients with hereditary spastic paraplegias type 5.Methods Eleven patients with SPG5 who underwent cervico-thoracic spinal cord MR examination and spastic paraplegia rating scale(SPRS)were prospectively enrolled.The second MR examination and SPRS were completed after 1 year follow-up,and the changes of SPRS score,the overall structures and microstructures of spinal cord were compared.Results No significant difference of SPRS scores was found(P＞0.05).Compared to the first cervico-thoracic spinal cord MR examination,atrophy of spinal cord aggravated in the second time MRI.Significant difference of axial diffusivity of C4 right corticospinal tract(CST)was found(t=3.987,P＜0.01),but not of the other parameters of C4(all P＞0.05)between the first and the second time MRI.No significant difference of fractional anisotropy(FA),mean diffusivity(MD),AD,radial diffusivity(RD)nor T1 value of the other centrums'white matter,posterior funiculus or bilateral CST in spinal cord was found between the first and the second time MRI(all P＞0.05).Meanwhile,no significant difference of CSA,left and right diameter nor anteroposterior diameter of C1-T9 was found between the first and second time MRI(all P＞0.05).FA value of white matter,posterior funiculus and bilateral corticospinal tract in cervical spinal cord were all lower,whereas RD value at the above position were all higher in the second time MRI than those in the first time MRI(all P＞0.05).Conclusion Multiparameter MRI could be used for follow-up observation on changes of microstructure spinal cord in patients with hereditary spastic paraplegias type 5.

Objective:To investigate the feasibility of using spectral CT chest enhancement venous phase images to obtain virtual non-contrast (VNC) images and virtual arterial phase images to achieve one phase scan instead of three phase scans.Methods:Imaging data of 100 patients who underwent spectral CT chest plain and dual-phase enhancement scans at the First Affiliated Hospital of Kunming Medical University from January to May 2022 were analyzed retrospectively. The venous phase images of all patients were post-processed to obtain virtual non contrast (VNC) and 40 keV virtual mono-energy images (VMI) for simulated arterial phase images (Vart). Image quality and lesion detection were compared between true non-contrast (TNC) and VNC images, and conventional arterial phase (CIart) and Vart images by paired t-test, Wilcoxon test and McNemar test. Objective evaluation indexes of image quality included thoracic aorta, all levels of pulmonary artery, T4 vertebral bone cancellous, chest wall fat CT value, background noise, signal-to-noise ratio (SNR), and contrast-to-noise ratio (CNR); the subjective score of image quality was assessed by double-blind method on a 5-point scale; the assessment of lesion detection included the detection rate of calcified foci and pulmonary nodules size, density, volume, percentage of solid components and imaging characteristics. Results:Except for chest wall fat and T4 vertebral bone cancellous, the differences in CT values between VNC and TNC images were not statistically significant ( P>0.05); Except for right upper pulmonary artery and right lower pulmonary artery, the differences in background noise values and SNR between TNC images and VNC images were statistically significant ( P<0.05). Compared with CIart images, the CT values of chest wall fat were lower in Vart images ( P<0.05), and the CT values of the remaining sites were significantly higher ( P<0.05); the background noise values of all sites on Vart images were smaller than those on CIart images, and the SNR and CNR values were higher than those on CIart images ( P<0.05).The differences of subjective scores of images quality were not statistically significant between TNC and VNC images, between CIar and Vart images ( P>0.05). Taking TNC as the standard, the overall detection rate of VNC for calcification was 88.53% (301/340). Except for the short diameter of partially solid nodules, the differences of qualitative and quantitative assessment indexes of lung nodules between TNC and VNC images were not statistically significant ( P>0.05). Conclusions:Spectral CT chest venous phase 40 keV VMI can simulate arterial phase images in the diagnosis of vascular lesions, and venous phase VNC basically meets the standard of conventional plain scan, so it is feasible to replace plain scan, arterial phase and venous phase images in a certain range.

This paper summarized professor ZHANG Lei′s experience in treating scleroderma from “extraordinary pathogens entering collaterals”. The basic pathogenesis of scleroderma is “extraordinary pathogens entering the collaterals”， and extraordinary pathogens can be divided into external and internal categories. External extraordinary pathogens are mostly exogenous wind， cold and damp pathogens， and the pathogenesis is wind， cold and damp invading skin stria and retaining collaterals. Most of the endogenous extraordinary pathogens are turbid phlegm and blood stasis， and the pathogenesis is endogenous phlegm and stasis leaving the channels and overflowing the collaterals， and blocking the collaterals. Blocked by extraordinary pathogens for a long time， the long illness will lead to deficiency and develop into a syndrome of collaterals excess and channels deficiency. Therefore， professor ZHANG creats Tengluo Beverage （藤络饮） as the basic formula to unblock collaterals and dispel pathogens， and recommends to add or subtract it according to the different syndrome and pathogenic characteristics of the edema stage， sclerosis stage， and atrophy stage. In the edema stage， it is advised to expel wind， remove dampness and unblock the collaterals， while in the sclerosis stage， the method of dissolving phlegm， expelling stasis and unblocking collaterals should be used； in the atrophic stage， it is suggested to differentiate the deficiency of qi， blood， yin and yang， and eliminate extraordinary pathogens on the basis of reinforcing healthy qi .

Objective To investigate the clinical effect of free posterior interosseous artery perforator flap combined with muscle fascia for repairing soft tissue and extensor tendon defect of hand.Methods Fifteen cases of hand skin soft tissue and extensor tendon defect admitted to Ningbo No.6 Hospital from December 2017 to December 2020 were repaired with free posterior interosseous artery perforator flap combined with extensor carpi ulnaris muscle fascia transplantation,and curative effect was observed.Results All flaps survived and patients were followed up for 6-24 months.The texture and thickness of the flap were satisfactory,and the recovery of the finger extension and flexion function were good.The excellent and good rate of hand tendon repair was 66.7%.In three cases with nerve anastomosis,the skin flap sensation recovered to S3.Conclusion The free posterior interosseous artery perforator flap combined with muscle fascia has a good clinical effect in repairing hand skin soft tissue and tendon defect.

Objective:Using the information technology of knowledge element theory to analyze the content of the ancient Chinese medicine book Nv Ke Xin Fa, and to explore the original thinking and implicit knowledge of the author of the book Nv Ke Xin Fa. Methods:Based on the theory and technology of knowledge element, the indexing data was read as logical data by using MS SQL Server database and then the knowledge graph of traditional Chinese medicine was constructed by using neo4j database.Results:There are 345 knowledge bodies and 1 478 knowledge elements in the Nv Ke Xin Fa, mainly including prescription name (280), composition (275) and usage (224); There are 711 semantic types, mainly including prescriptions (232), etiology and pathogenesis (221), syndrome manifestations (125) and treatment (88); 187 semantic associations, mainly syndrome differentiation and treatment (172); Knowledge of semantic association.There are 34 body types, with leucorrhea (16), amenorrhea (13), blood Avalanche (13) and asthenia (13). Conclusions:Zheng Qinyu paid attention to distinguishing the texture of leucorrhea in the treatment of leucorrhea, put forward the pathogenesis and treatment principles and methods of leucorrhea in the stages of "before childbirth" and "after childbirth", and made it clear that the application scope of Buzhong Yiqi Decoction involves menstruation, leucorrhea, pregnancy and the whole cycle of childbirth.Mrs Zheng Qinyu pays great attention to the protection on the spleen and stomach of the middle coke and regulation of Qi that could rise and fall, which could provide a reference for modern clinical diagnosis and treatment of leukorrhagia diseases.

Objective : The purpose of this study was to investigate the effect of different exposure sequence and interval on the post-antibiotic effect(PAE) and 24-hour time-kill produced by linezolid and fosfomycin in combination againstStaphylococcus aureus(S.aureus). Methods: PAEs were evaluated by exposingS.aureusto linezolid and fosfomycin individually and simultaneously for 1 h.Further assess of PAEs with the organism exposed first to linezolid for 1 h, followed by a second 1 h fosfomycin exposure at the beginning(Tb),middle(Tm)and end(Te) of the PAE phase induced by linezolid.The next study was similar to the above, but with the sequence of the two antibiotics reversed, i.e.fosfomycin then linezolid.The 24-hour time-kill studies were performed with three exposure regimens: simultaneous exposure of the two antibiotics; first exposure to linezolid for 1 h and then combined with fosfomycin, and first exposure to fosfomycin for 1 h and then combined with linezolid to draw the sterilized curve. Results: Compared with the PAE of individual antibiotics [ linezolid(1.2±0.1) h, fosfomycin(1.6±0.1) h],the PAE produced by simultaneous exposure of the combination(2.9±0.1) h presented additive interaction.However, the PAE of fosfomycin decreased or even disappeared(-0.3 h for Tb, 0 h for Tm and 0.9 h for Te) during the PAE phase of linezolid, and with a stronger antagonism at the beginning.Differently, an additive interaction was observed again when the test organism was exposed to fosfomycin first.In the 24-hour time-kill assays, the antibacterial effect of the first exposure to fosfomycin regimens was significantly stronger(difference of 1 log10)than that of the simultaneous exposure and the first exposure to linezolid, while the latter two were similar. Conclusion The pharmacodynamic behaviour of linezolid combined with fosfomycin againstS.aureuswas influenced by exposure regimen.The first exposure to fosfomycin regimens rather than simultaneous exposure may exert the optimal antibacterial activity.Therefore, the exposure regimen is one of the important factors to be considered for the clinical application of this combination.

Objective To investigate the effects of ketogenic amino acid ( KAA) replacement diet on insulin resistance in mice fed with high fat diet(HFD) and to analyze the possible mechanism. Methods C57BL mice were fed with a control diet, HFD, and KAA-fortified HFD(HFDKAAR)from the age of 8 weeks, and 8 weeks after HFD initiation, the HFD-fed mice were divided into two groups:one group of mice were fed the same HFD, the other group were fed HFDKAAR ( HFD→HFDKAAR ) . The metabolic evaluations were performed at the end of 16 weeks. Blood glucose levels were measured at 0, 15, 30, 60, 90, and 120 min after the injection of glucose ( 1 g/kg BW intraperitoneal glucose tolerance test, ipGTT) . The insulin,β-hydroxybutyrate, and acetoacetate levels in the plasma were measured via ELISA. The insulin resistance index ( IRI) and area under curve ( AUC) were calculated. The expression of hepatic LKB1 ( liver kinase B1 ) , AMP-activated protein kinase ( AMPK ) , and mTOR ( Mammalian target of rapamycin ) protein, and mcp-1 mRNA were measured by western blot and real-time PCR respectively. Results HFD-fed group of mice displayed significantly heavier body weight,heavier intra-abdominal fat weight, and significant deterioration of glucose tolerance at the end of 16 weeks in addition to higher insulin levels( all P<0. 05), HFDKAAR-fed mice exhibited significantly ameliorated high fat diet-induced obesity and glucose intolerance compared to the HFD-fed mice, which was associated with decreased insulin levels, IRI, AUC, and mcp-1 mRNA expression (all P<0. 05). HFD suppressed hepatic LKB1 and AMPK phosphorylation expression, and increased mTOR phosphorylation levels compared to the control diet-fed mice(all P<0. 05). In contrast, treatment with the HFDKAAR diet increased LKB1and p-AMPK expression, which was associated with suppressed p-mTOR levels compared to the HFD-fed mice(all P<0. 05). Conclusion KAA may ameliorate high fat diet-induced obesity, glucose intolerance, via normalizing the hepatic LKB1-AMPK-mTOR nutritional signal passageway. KAA replacement diet seems to be a potential nutritional intervention for the treatment for patients with metabolic defects, such as obesity, glucose intolerance, as well as metabolic syndrome.

Objective To study the effects of postnatal overfeeding and high-fat diet on blood pressure of rats,and to explore the pathophysiological mechanism underlying hypertension induced by continuous early postnatal overfeeding.Methods Male Sprague-Dawley rats were randomly divided into normal feeding group (10/litter) and overfeeding group (3/litter) on postnatal day 3 with a random number table.After weaning at postnatal week 3,the rats were randomly given standard chow or high-fat (HF) diet until week 16.Hence four groups were analyzed,namely normal feeding group,breastfed overfeeding group,post-weaning overfeeding group,and continuous overfeeding group.Body weight was continuously monitored in each week.Visceral fat pad (retroperitoneal and perigenital),systolic pressure,and heart rate were observed at week 3 and week 16.Thoracic aorta was sampled for measurement of vascular endothelial dilation function.Histological morphology was observed with HE staining,nitric oxide content of thoracic aorta was detected with nitrate reductase method.The mRNA expression of endothelial nitric oxide synthase (eNOS) in thoracic aorta was determined by real-time polymerase chain reaction.The protein expressions of eNOS and phosphorylated eNOS were determined by Western blot.Results At week 3,breastfed overfeeding rats displayed significantly larger body weight [(77.80 ± 0.57) g vs.(62.80 ±0.85) g,t =14.576,P ＜ 0.01] and visceral fat [retroperitoneal:(8.19 ± 0.49) mg/g vs.(4.92 ± 0.31) mg/g,t =5.629,P＜0.01;perigenital:(3.50 ±0.29) mg/g vs.(2.08 ±0.13) mg/g,t =4.552,P ＜0.01] compared with normal feedindg rats,and the protein expression of phosphorylated eNOS in aortic tissues was significantly reduced to week 16 (F =15.215,P ＜0.01);high-fat diet feeding after weaning further increased the body weight and fat mass in breastfed overfeeding rats.At week 16,continuous overfeeding rats showed hypertension [(149 ± 1.94) mmHg (1 mmHg =0.133 kPa),F =22.834,P ＜0.01],impaired vascular endothelial dilation function (F =7.648,P ＜ 0.05),and reduced protein expression of phosphorylated eNOS (F =15.215,P ＜ 0.01),while the post-weaning overfeeding group only had elevated blood pressure.Conclusions Overfeeding in breastfeeding period and high-fat diet after weaning leads to hypertension.The continuous decrease in phosphorylated eNOS in vascular tissues may be an important molecular process participating in the occurrence of vascular endothelial dysfunction in adults induced by postnatal overfeeding.

Aim To study the mechanism of 8-isopro-pylaminomethyl hesperitin ( IPHP ) intestinal absorp-tion using Caco-2 cell lines. Methods Using Caco-2 cell lines as an intestinal epithelial cell model, the effects of drug concentration, temperature, pH, P-gly-coprotein ( P-gp) inhibitor verapamil and multidrug re-sistance protein 2 ( MRP2 ) inhibitors MK-571 or pro-benecid on IPHP transport across Caco-2 cell lines were all investigated. Results The transportation of IPHP was related to drug concentration. The Papp ( AP-BL) ( × 10 -5) was (2. 21 ± 0. 200) cm·s-1,(3. 56 ± 0. 306) cm·s-1,(3. 81 ± 0. 179) cm·s-1,(4. 23 ± 0. 229 ) cm · s-1 , ( 4. 17 ± 0. 262 ) cm · s-1 , re-spectively, and Papp(BL-AP) ( × 10 -5) was (3. 57 ±0. 209) cm·s-1,(4. 51 ± 0. 113) cm·s-1,(4. 97 ± 0. 229) cm·s-1,(5. 24 ± 0. 550) cm·s-1,(5. 07 ± 0. 557) cm·s-1,respectively. Efflux rate was 1. 61, 1. 26,1. 3,1. 23,1. 21,respectively. Temperature and pH both influenced the transport, While the P-gp in-hibitor verapamil had no effect on the transport of IPHP. MRP2 inhibitors MK-571 or probenecid led to an apparent decrease in the efflux of IPHP. Conclu-sion The results suggest that the transport of IPHP is mainly passive diffusion, and MRP2 but not P-gp may be involved in the transport of IPHP.