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@#Abstract：Objective To carry out serological analysis of varicella⁃zoster virus（VZV）IgG antibody level in healthy people aged 1 ~ 30 years in Liaoning Province. Methods In October 2020，3～5 mL venous blood samples were collected from 617 healthy people aged 1～30 years selected from six counties and districts in Shenyang，Fuxin and Dandong of Liaoning Province by stratified random sampling method，of which serum samples were collected and determined for VZV IgG antibody level by ELISA. The positive rate of serum antibody and geometric mean concentration（GMC）of antibody were calculated and compared. Results Among 617 serum samples，302 samples were positive for VZV IgG antibody，the positive rate was 48. 947%，and the GMC was 112. 772 mIU／mL. The positive rate of VZV IgG antibody was 29. 670%～75. 789% and the GMC was 45. 508～366. 559 mIU／mL in healthy people of various ages. Both of the antibody positive rate（χ2 = 67. 104， P < 0. 001）and GMC（F = 20. 685，P < 0. 001）showed significant differences. The positive rates of VZV IgG antibody in male and female were 44. 817% and 53. 633% respectively，which showed significant difference（χ2 = 4. 779，P = 0. 029）， while the GMCs were 96. 983 and 133. 829 mIU／mL respectively（t = -1. 958，P = 0. 051）with no significant difference. The positive rates of VZV IgG antibody of healthy people in Shenyang，Fuxin and Dandong of Liaoning Province were 55. 224%，40. 201% and 51. 152% respectively with significant differences（χ2 = 9. 683，P = 0. 008），of which the positive rate of FuxinwassignificantlylowerthanthoseofShenyangandDandong（χ2 =9. 046and5. 013，P =0. 003and0. 025，respectively）； While the GMCs were 133. 523，85. 953 and 123. 713 mIU／mL respectively with no significant difference（F = 0. 514， P = 0. 598）. Among 617 serum samples，54 sampleswere suspicious，which remained within the criticalrange afterre⁃examina⁃ tion，while the gap between positive rate and the total percentage of positive and suspicious results gradually decreased with the increase of age，indicating that the immunity to varicella gradually increased with the increase of age. Conclusion The VZV⁃IgG antibody level of healthy people aged 1～30 years in Liaoning Province increased gradually with age，while the overall level was low. To control the spread of varicella virus，it is recommended to increase varicella vaccine coverage in vulnerable areas and susceptible population to build VZV immune barrier.
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@#Abstract: Objective By collecting and sorting the information of varicella cases reported in Liaoning Province from 2006 to 2021, the epidemiological characteristics were analyzed, and the monthly incidence data were predicted, so as to explore the prevention and control strategy of varicella disease in Liaoning Province. Methods By collecting the characteristic information of varicella cases in Liaoning Province, epidemiological analysis was carried out on the regional, population, and temporal characteristics of varicella incidence. The monthly incidence data of varicella were fitted with Eviews software, seasonal ARIMA model was used for modeling, and models were selected according to SC and AIC. After modeling, the model was used to predict the incidence data in 2022. Results The incidence rate of varicella in Liaoning Province has increased in recent years. The onset time was "bimodal distribution", with the main peak occurring from November to January of the next year and the secondary peak occurring from May to June. Since 2019, the onset age has shifted backward. From the original 0-<10 age group with the highest incidence rate, it shifted to the 10-<20 age group with the highest incidence rate. From 2006 to 2021, the incidence of varicella mainly concentrated in people aged 0 to <40 years old, and the incidence rate of the population over 40 years old showed a cliff-like decline. The incidence of chickenpox was higher in the central region of Liaoning Province, such as Shenyang, Dalian, Anshan and Panjin, and relatively low in Huludao, Jinzhou, Fuxin and Liaoyang. The distribution of the population was mainly students, followed by kindergartens and scattered children. ARIMA model of monthly incidence data was established by software as ARIMA (1, 0, 1) (1, 1, 1)12. Conclusions The incidence rate of varicella in Liaoning Province has been rising in recent years. The incidence is obviously seasonal, and the age group of the affected population has moved backward. It is predicted that the incidence will continue to increase in 2022. The prevention and control of varicella should still be the current key work. In order to reduce the population incidence rate, two-dose vaccination strategies should be vigorously promoted the implementation of the, and the inclusion of varicella vaccine in the immunization program should be achieved as soon as possible.
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Objective To improve the fixation method of the transmission electron microscope for better morphological preservation of mitochondria and lipid droplets in mouse brown adipose tissue. Methods The fixation method for mouse brown adipose tissue was optimized, mainly including an increased concentration of paraformaldehyde from 2% to 4% in the pre-fixative, employment of transcardial perfusion followed by immersion fixation in pre-fixation, and using imidazole-buffered osmium tetroxide as the post-fixative. The ultrastructures of brown adipocytes prepared by the improved method were observed and compared with those of a known standard protocol (3 mice in each group). The improved method was further validated in the quantitative analysis of mitochondrial cristae density and lipid droplets. Results The mitochondrial cristae and membrane structure of other organelles of brown adipocytes were better preserved using the optimized method compared with those of the standard method. Lipid droplets were presented as round structures with high electron density instead of vacuolated appearances. Using this method, we observed that the density of mitochondrial cristae and the content of lipid droplets increased in brown adipocytes after cold adaptation. Conclusion The optimized method can better preserve the ultrastructure of organelles in brown adipocytes, especially mitochondria and lipid droplets, and ma)' be applicable for studying the ultrastructures remodeling of brown adipose tissue under different physiological or pathological conditions.
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OBJECTIVE@#To observe the effect of wheat-grain moxibustion at "Dazhui" (GV 14) on the expressions of Beclin-1 and GRP78 in spinal dorsal horn in rats with cervical spondylotic radiculopathy (CSR), and to explore the possible analgesic mechanism of wheat-grain moxibustion for CSR.@*METHODS@#A total of 48 SD rats were randomly divided into a sham operation group, a model group, a wheat-grain moxibustion group and a wheat-grain moxibustion+3-MA group, 12 rats in each group. The CSR model was prepared by spinal cord insertion method. Three days after modeling, the rats in the model group were intraperitoneally injected with 1 mL of 0.9% sodium chloride solution; the rats in the wheat-grain moxibustion group were treated with wheat-grain moxibustion at "Dazhui" (GV 14, 6 cones per time) on the basis of the model group; the rats in the wheat-grain moxibustion+3-MA group were intraperitoneally injected with 3-MA solution and wheat-grain moxibustion at "Dazhui" (GV 14, 6 cones per time). The three groups were intervened for 7 days, once a day. The gait score and mechanical pain threshold were observed before treatment and 7 days into treatment; after the treatment, the expressions of mRNA and protein of Beclin-1 in spinal dorsal horn were detected by real-time fluorescence quantitative PCR and immunohistochemistry; the expression of GRP78 protein in spinal dorsal horn was detected by Western blot method; the autophagosomes and ultrastructure in spinal dorsal horn neurons were observed by electron microscope.@*RESULTS@#After the treatment, compared with the sham operation group, in the model group, the gait score was increased and the mechanical pain threshold was decreased (P<0.01), and the expression of GRP78 protein in spinal dorsal horn was increased (P<0.01). Compared with the model group and the wheat-grain moxibustion+3-MA group, in the wheat-grain moxibustion group, the gait score was decreased and mechanical pain threshold was increased (P<0.01), and the expression of GRP78 protein in spinal dorsal horn was decreased, and the expressions of mRNA and protein of Beclin-1 were increased (P<0.01). Under electron microscope, the ultrastructure of spinal dorsal horn neurons in the wheat-grain moxibustion group was not significantly damaged, and its structure was basically close to normal, and the number of autophagosomes was more than the other three groups.@*CONCLUSION@#Wheat-grain moxibustion at "Dazhui" (GV 14) has analgesic effect on CSR rats. The mechanism may be related to moderately up-regulate the expression of Beclin-1, enhance autophagy and reduce endoplasmic reticulum stress.
الموضوعات
Animals , Rats , Beclin-1/genetics , Endoplasmic Reticulum Chaperone BiP , Moxibustion , RNA, Messenger , Radiculopathy/therapy , Rats, Sprague-Dawley , Spinal Cord , Spinal Cord Dorsal Horn , Spondylosis , Triticum/geneticsالملخص
This study explored the usefulness of two-dimensional shear wave elastography (2D-SWE) in the early assessment of corpora cavernosa fibrosis (CCF). New Zealand male rabbits were randomly assigned to an experimental group or a control group. Recombinant human transforming growth factor beta 1 (TGF-β1) was injected into the dorsal penis tissue of rabbits in the experimental group. Conventional ultrasound and 2D-SWE examinations were performed before and 20 days after injection. Penile histological analysis was performed by hematoxylin-eosin staining, sirius red staining, and immunohistochemistry. Measurement of 2D-SWE examination results was performed using shear wave elastography quantitative measurement (SWQ). Histological analysis outcomes were the proportion of smooth muscle cells (SMCs), collagen fibers (CFs), collagen type I (Col I), and collagen type III (Col III), as well as the SMCs/CFs ratio, measured by sirius red staining. Other histological analysis outcomes were the positive area proportion (PAP) of TGF-β1 (PAPT), fibronectin (PAPF), and Col III (PAPC), measured by immunohistochemistry. After recombinant human TGF-β1 injection, SWQ was higher in the experimental group than that in the control group (P < 0.001); however, there were no differences in conventional ultrasound results. There were significant differences in histological outcomes between the two groups (all P < 0.05). These results indicated that 2D-SWE was superior for identifying early histological changes in CCF.
الموضوعات
Animals , Male , Rabbits , Elasticity Imaging Techniques/methods , Fibrosis , Penis/pathology , Transforming Growth Factor beta1/metabolismالملخص
OBJECTIVE@#To compare the clinical effect of acupuncture combined with wheat-grain moxibustion and oral sertraline hydrochloride dispersible tablets in the treatment of mild to moderate postpartum depression.@*METHODS@#Sixty patients with mild to moderate postpartum depression were randomly divided into an observation group and a control group, 30 cases in each group. Both groups were treated with psychotherapy. The control group was treated with oral sertraline hydrochloride dispersible tablets, 50 mg each time, once a day; the observation group was treated with acupuncture at Qihai (CV 6), Zusanli (ST 36), Xuehai (SP 10), Hegu (LI 4), Sanyinjiao (SP 6), Taixi (KI 3), etc. combined with wheat-grain moxibustion at Xinshu (BL 15), Pishu (BL 20), Ganshu (BL 18) and Shenshu (BL 23), once every other day, 3 times a week. Both groups were treated for 4 weeks as a course, with 2 consecutive courses of treatment. Before and after treatment and follow-up of 3 months after the end of treatment, the Hamilton depression scale (HAMD), Edinburgh postnatal depression scale (EPDS) and World Health Organization quality of life-BREF (WHOQOL-BREF) score of the two groups were compared, and the clinical effect was assessed.@*RESULTS@#After treatment and during follow-up, the HAMD and EPDS scores of the two groups were lower than before treatment (@*CONCLUSION@#Acupuncture combined with wheat-grain moxibustion can improve the depressive symptoms of patients with mild to moderate postpartum depression and improve their quality of life, and the clinical effect is more lasting and stable than oral sertraline hydrochloride dispersible tablets.
الموضوعات
Female , Humans , Acupuncture Points , Acupuncture Therapy , Depression, Postpartum/therapy , Moxibustion , Quality of Life , Treatment Outcome , Triticumالملخص
One new and two known dammarane-type saponins were isolated from the leaves of Gynostemma pentaphyllum using various chromatographic methods. Their structures were identified by HR-ESI-MS,~( 1)H-NMR, ~(13)C-NMR, 2 D-NMR spectra as 2α,3β,12β,20,24(S)-tetrahdroxydammar-25-en-3-O-[β-D-glucopyranosyl(1→2)-β-D-glucopyranosyl]-20-O-β-D-xylopyranosyl(1→6)-β-D-glucopyranoside(1, a new compound, namely gypenoside J5) and 2α,3β,12β,20,24(R)-tetrahdroxydammar-25-en-3-O-[β-D-glucopyranosyl(1→2)-β-D-glucopyranosyl]-20-O-β-D-xylopyranosyl(1→6)-β-D-glucopyranoside(2) and 2α,3β,12β,20-tetrahydroxy-25-hydroperoxy-dammar-23-en-3-O-[β-D-glucopyranosyl(1→2)][β-D-glucopyranosyl]-20-O-[β-D-xylopyranosyl(1→6)]-β-D-glucopy-ranoside(3), respectively. Compounds 1 and 2 were a pair of C-24 epimers. All compounds showed weak cytotoxicity agxinst H1299, HepG2, PC-3, SH-SY5 Y cancer cell lines. However, they exerted protective effect against SH-SY5 Y cellular damage induced by H_2O_2 dose-dependently, of which compound 1 displayed the strongest antioxidant effect. The present study suggested that G. pentaphyllum has antioxidative potential and the saponins from G. pentaphyllum are considered as the active compounds with neuroprotecitve effect.
الموضوعات
Gynostemma , Molecular Structure , Neuroprotective Agents/pharmacology , Saponins/pharmacology , Triterpenes/pharmacologyالملخص
Objective:To evaluate the effect of ulinastatin on hyperoxia-induced acute lung injury (ALI) and its relationship with Wnt/β-catenin signaling pathway in infantile rats.Methods:A total of 36 clean-grade Sprague-Dawley rats, aged 14 days, weighing 40-50 g, were divided into 3 groups ( n=12 each): control group (C group), hyperoxia-induced ALI group (ALI group) and ulinastatin group (UTI group). Hyperoxia-induced ALI was induced by inhaling oxygen at concentration greater than 90% for 72 h. At 1 day after the model was established successfully, ulinastatin 50 000 U/kg was injected intraperitoneally daily at the same time for 3 consecutive days in group UTI, while the equal volume of normal saline was injected intraperitoneally at the same time point in C and ALI groups.The animals were sacrificed at 4 days after the model was established successfully, the lung tissues were taken for determination of the wet/dry weight ratio (W/D ratio), for microscopic examination of the pathological changes which were scored, for measurement of interleukin-6 (IL-6) IL-1β and tumor necrosis factor-alpha (TNF-α) (by enzyme-linked immunosorbent assay) and for detection of the expression of phosphorylated glycogen synthase kinase (p-GSK-3β), Wnt3a and β-catenin by Western blot, and ultrastructure was examined with with an electron microscope. Results:Compared with C group, W/D ratio and lung injury score were significantly increased, the contents of IL-6, IL-1β and TNF-α were increased, and the expression of p-GSK-3β, Wnt3a and β-catenin were up-regulated in lung tissues in group ALI ( P<0.05). Compared with group ALI, W/D ratio and lung injury score were significantly decreased, the contents of IL-6, IL-1β and TNF-α were decreased, and the expression of p-GSK-3β, Wnt3a and β-catenin were down-regulated in lung tissues in group UTI ( P<0.05). The ultrastructure injury in group UTI was reduced as compared with group ALI. Conclusion:The mechanism by which ulinastatin can alleviate hyperoxia-induced ALI is related to inhibiting the activation of Wnt/β-catenin signaling pathway and decreasing inflammatory response in infantile rats.
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Objective:To evaluate the role of histone deacetylase 6 (HDAC6) in spinal dorsal horn in dexmedetomidine-induced reduction of neuropathic pain (NP) in rats.Methods:Forty clean-grade healthy male Sprague-Dawley rats, aged 7-9 weeks, weighing 190-240 g, were divided into 5 groups ( n=8 each) using a random number table method: control group (group C), sham operation group (group SH), group NP, dexmedetomidine group (group D), and specific HDAC6 inhibitor ACY-1215 plus dexmedetomidine group (group AD). The animals were commonly fed without any treatment in group C. The sciatic nerve was only isolated but not ligated in group SH.The animals were anesthetized with intraperitoneal 10% chloral hydrate 350 mg/kg.NP was induced by chronic constrictive injury (CCI). The right sciatic nerve was exposed, and 4 loose ligatures were placed on the sciatic nerve at 1-mm intervals with 4-0 silk thread in NP and D groups.In group D, dexmedetomidine 40 μg/kg was intraperitoneally injected once a day starting from the end of operation until the animals were sacrificed.In group AD, ACY-1215 25 mg/kg was intraperitoneally injected every day immediately before CCI, and dexmedetomidine 40 μg/kg was intraperitoneally injected daily after CCI until 15 days after CCI.The equal volume of solvent was given instead of dexmedetomidine in S and NP groups.The mechanical paw withdrawal threshold to von Frey filament stimulation (MWT) and thermal paw withdrawal latency (TWL) were measured at 1 day before CCI (baseline, T 0) and 3, 6, 9, 12 and 15 days after CCI (T 1-5). The rats were then sacrificed, and the dorsal horn tissues of L 4-6 spinal cord were obtained for determination of the expression of myeloid differentiation factor 88 (MyD88) and nuclear factor kappa B (NF-κB) p65 by Western blot. Results:Compared with group C and group SH, the MWT was significantly decreased, and the TWL was shortened at T 1-5, and the expression of MyD88 and NF-κB p65 in the spinal dorsal horn was up-regulated in NP, D and AD groups ( P<0.05). Compared with group NP, the MWT was significantly increased, and the TWL was prolonged at T 1-5, the expression of MyD88 and NF-κB p65 in the spinal dorsal horn was down-regulated in group D ( P<0.05), and no significant change was found in the parameters mentioned above in group AD ( P>0.05). Compared with group D, MWT was significantly decreased, and TWL was shortened at T 1-5, and the expression of MyD88 and NF-κB p65 was up-regulated in the dorsal horn of the spinal cord in group AD ( P<0.05). Conclusion:HDAC6 in spinal dorsal horn is involved in dexmedetomidine-induced reduction of NP in rats, which is related to inhibiting MyD88/NF-κB signaling pathway.
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Objective:To investigate the effects of isoflurane anesthesia on the transcription levels of clock genes RORα and Rev-erbα and their circadian rhythm changes in the hippocampus and cerebral cortex of mice. Methods:Thirty-six 7-week-old male C57BL/6J mice were divided into control group and isoflurane anesthesia group ( n=18) according to random number table method. They were subjected to 12 h light/12 h dark environment for 1 week. Mice in the isoflurane anesthesia group were anesthetised with 1.2% isoflurane at 22:00 pm on the night before the experiment for 6 h; 4 h after anesthesia, the light time of 8:00 am was taken as the zeitgeber time (ZT0), and then, every 8 h was taken as the recorded time (ZT8, ZT16, ZT24, ZT32 and ZT40). The RORα and Rev-erbα mRNA expressions were measured by real-time fluorescent quantitative PCR (RT-qPCR) at each time point in the cerebral cortex and hippocampus of the two groups. The cosine curves of RORα and Rev-erbα mRNA expressions were analyzed by Chronos-Fit software. Results:As compared with the control group, the isoflurane anesthesia group had significantly down-regulated RORα mRNA expression in the hippocampus, significantly up-regulated RORα mRNA expression in the cerebral cortex, and significantly down-regulated Rev-erbα mRNA expression in the cerebral cortex ( P<0.05). The RORα and Rev-erbα mRNA expressions in the hippocampus and cerebral cortex of the control group showed a rhythmic cycle of about 24 h; while the circadian rhythm of RORα mRNA shifted to the right by 5.41 h, and the circadian rhythm of Rev-erbα mRNA shifted to the left by 0.89 h in the hippocampus of the isoflurane anesthesia group. The peak circadian rhythm of RORα mRNA shifted to the right by 0.35 h, and the peak circadian rhythm of Rev-erbα mRNA shifted to the left by 0.56 h in the cerebral cortex of the isoflurane anesthesia group. Conclusion:Isoflurane anesthesia can induce the changes of RORα and Rev-erbα transcription levels and circadian rhythm changes in the hippocampus and cerebral cortex of mice.
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4-IBB and 4-IBB ligand (4-1BBL) , also known as CI) 137 and CD 137 ligand, are members of tumor necrosis factor (TNF) receptor and ligand family, respectively.The interaction of 4-1BBL and 4-IBB can activate T cell immune response.Therefore, 4-1BBL has been playing a role as a classical costimulatory molecule involving in anti-tumor immune responses.In recent decades, it was reported that 4-1BBL had multiple functions in tumor cells.However, the molecular mechanism of 4-1BBL in the progression of gastric cancer (GC) remains unclear.This study investigated the biological function and molecular mechanism of 4-1BBL in human GC cells.First, analysis from TCGA and Kaplan Meier plotter databases showed that 4-1BBL expression level in GC tissues was significantly higher than that in adjacent tissues (P<0.001) , and 4-1BBL high expression was positively associated with poor prognosis of GC (P<0.05).Knockout of 4-1BBL significantly inhibited the proliferation (P<0.05) , invasion and migration of GC cells (P<0.05 ) , and increased the apoptosis of GC cells (P<0.05).Western blot showed that 4- 1 BBL knockout decreased the protein expression levels of (3-catenin, c-Myc and Cyclin D1, and blocked Wnt/p-catenin signaling pathway.On the contrary, 4-1 BBL overexpression significantly promoted the proliferation (P<0.05) , invasion and migration of GC cells (P<0.05) , and reduced the apoptosis of GC cells (P<0.05).Moreover, 4-1 BBL overexpression increased the protein expression levels of (3- catenin, c-Myc and Cyclin D1, and activated Wnt/p-catenin signaling pathway.In summary, 4-1 BBL promoted the proliferation and migration of human GC cells through Wnt/(B-catenin signaling pathway.
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CRISP R/Cas9 is an emerging gene editing technique, which plays an important role in life science research.It is of great significance to introduce this cutting-edge scientific technique into the experimental teaching for undergraduates.Therefore, we established an undergraduate experiment system based on CRISPR/Cas9 technology.This experiment system focuses on the application of CRISPR/Cas9- mediating gene editing in mammalian cells.An engineered mouse embryonic fibroblast which genome were inserted with fluorescein mCherry gene was selected as the experimental model, and called STO-82.Firstly, sgRNAs targeting mCherry gene were designed to construct CRISPR-Cas9/sgRNA co-expression plasmids.After being confirmed by sequencing, they were transfected into STO-82 cells.Two groups of cells with mCherry negative and positive signals were detected by fluorescence-activated cell sorting.Single cells with negative fluorescence were separated and then cultivated to become monoclonal cells.The mutation status of mCherry gene in monoclonal cell lines was detected by sequence analysis.The result showed that there were mutations of insertion or deletion at target sites, indicating that the experimental system was successfully established.Therefore, this comprehensive experiment is comprised of sgRNA design, construction of CRISPR-Cas9/sgRNA co-expression plasmids, cell transfection, cell sorting, monoclonal cell cultivation and sequence analysis.This experiment system is used for experimental teaching for senior undergraduates.Teaching practice can either be decomposed into content modules or be taken as a whole program in light of actual situation.In the teaching practice at 3 classes (13 groups in total, two students every group), which adopted the model of small-class teaching (about 10 students per class), the majority completed the content modules and the expected outcomes were achieved.Through the design and teaching practice of this experiment system, the students acquire a deeper understanding for the principle and experimental procedure of CRISPR/Cas9 technology, an enhanced experimental ability and rigorous scientific thinking and also some knowledge in the risk of its medical application.
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Objective To evaluate the effect of penehyclidine hydrochloride (PHC) on Toll-like receptor 4 (TLR4) /nuclear factor kappa B (NF-κB) signaling pathway in non-ventilated lung injury in the patients undergoing radical operation for lung cancer.Methods A total of 100 patients,aged 40-64 yr,with body mass index 18-27 kg/m2,of American Society of Anesthesiology physical status Ⅱ or Ⅲ,undergoing radical operation for lung cancer,were divided into 2 groups (n=50 each) according to the random number table method:control group (group C) and PHC group.PHC 0.01 mg/kg was intravenously injected at 10 min before anesthesia induction in group PHC,while the equal volume of normal saline was given instead in group C.The peripheral tissues of the removed lung tissues were obtained for determination of wet/dry weight ratio (W/D ratio).The pathological changes and ultrastructure of lung tissues were observed under light microscope,and lung injury was assessed and scored.The expression of TLR4 and NF-κB protein and mRNA was detected by Western blot and real-time polymerase chain reaction,respectively.Before administration (T0),at the onset of one-lung ventilation (T1),at 60 min of one-lung ventilation (T2),immediately after the end of one-lung ventilation (T3),at the end of operation (T4) and at 24 h after operation (T5),blood samples were collected from the internal jugular vein for determination of serum tumor necrosis factor-alpha,interleukin-6 (IL-6) and IL-8 concentrations by enzyme-linked immunosorbent assay.Results Compared with group C,the W/D ratio and lung injury scores were significantly decreased,the expression of TLR4 and NF-κB protein and mRNA was down-regulated,and the concentrations of tumor necrosis factor-alpha,IL-6 and IL-8 were decreased at T12-T5 in group PHC (P＜0.05).The pathological changes and damage to ultrastructure of lung tissues were significantly attenuated in group PHC as compared with group C.Conclusion The mechanism by which PHC attenuates non-ventilated lung injury is related to blocking TLR4/NF-κB signaling pathway and reducing inflammatory responses in the patients undergoing radical operation for lung cancer.
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Objective@#To evaluate the effect of penehyclidine hydrochloride (PHC) on Toll-like receptor 4 (TLR4)/nuclear factor kappa B (NF-κB) signaling pathway in non-ventilated lung injury in the patients undergoing radical operation for lung cancer.@*Methods@#A total of 100 patients, aged 40-64 yr, with body mass index 18-27 kg/m2, of American Society of Anesthesiology physical status Ⅱ or Ⅲ, undergoing radical operation for lung cancer, were divided into 2 groups (n=50 each) according to the random number table method: control group (group C) and PHC group.PHC 0.01 mg/kg was intravenously injected at 10 min before anesthesia induction in group PHC, while the equal volume of normal saline was given instead in group C. The peripheral tissues of the removed lung tissues were obtained for determination of wet/dry weight ratio (W/D ratio). The pathological changes and ultrastructure of lung tissues were observed under light microscope, and lung injury was assessed and scored.The expression of TLR4 and NF-κB protein and mRNA was detected by Western blot and real-time polymerase chain reaction, respectively.Before administration (T0), at the onset of one-lung ventilation (T1), at 60 min of one-lung ventilation (T2), immediately after the end of one-lung ventilation (T3), at the end of operation (T4) and at 24 h after operation (T5), blood samples were collected from the internal jugular vein for determination of serum tumor necrosis factor-alpha, interleukin-6 (IL-6) and IL-8 concentrations by enzyme-linked immunosorbent assay.@*Results@#Compared with group C, the W/D ratio and lung injury scores were significantly decreased, the expression of TLR4 and NF-κВ protein and mRNA was down-regulated, and the concentrations of tumor necrosis factor-alpha, IL-6 and IL-8 were decreased at T2-T5 in group PHC (P<0.05). The pathological changes and damage to ultrastructure of lung tissues were significantly attenuated in group PHC as compared with group C.@*Conclusion@#The mechanism by which PHC attenuates non-ventilated lung injury is related to blocking TLR4/NF-κВ signaling pathway and reducing inflammatory responses in the patients undergoing radical operation for lung cancer.
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Four flavonoids were isolated from Gynostemma pentaphyllum by chromatography methods and their structures were identified by MS and NMR spectra data as quercetin-3-O-( 2″,6″-di-α-L-rhamnosyl)-β-D-galactopyranoside( 1),quercetin-3-O-( 2″,6″-di-α-L-rhamnosyl)-β-D-glucopyranoside( 2),quercetin-3-O-( 2″-α-L-rhamnosyl)-β-D-galactopyranoside( 3),and quercetin-3-O-( 2″-α-L-rhamnosyl)-β-D-glucopyranoside( 4). Among them,compounds 1-3 were obtained from the Cucurbitaceae family for the first time.The four flavonoids showed potent antioxidant effects against the DPPH,·OH and ■radicals in vitro,especially for DPPH radical scavenging activity with the IC50 values of 71. 4,29. 5,48. 3 and 79. 2 μmol·L~(-1),respectively. Moreover,the four flavonoids displayed strong cytoprotection against AAPH-induced oxidative damage in LLC-PK1 cells by suppressing the increase of malondialdehyde( MDA) and the decrease of the superoxide dismutase( SOD) and glutathione( GSH). Since further research is needed to prove its efficacy in vivo and clinical trial,the study may provide four potential antioxidants from G. pentaphyllum.
الموضوعات
Animals , Antioxidants , Flavonoids , Gynostemma , LLC-PK1 Cells , Oxidative Stress , Plant Extracts , Quercetin , Swineالملخص
Structural alterations in fibroelastic components of the penile corpus cavernousum (CC) may impair its compliance, resulting in venous leakage and erectile dysfunction (ED). Our study evaluated the effectiveness of noninvasive two-dimensional shear-wave elastography (2-D SWE) in quantifying penile CC lesions in rabbits with hyperlipidemia-induced ED. A total of 12 New Zealand white rabbits were randomly divided into two groups. Six were fed a high-cholesterol diet containing 2% cholesterol and 8.5% lard for 10 weeks and the other six were fed normal diet as controls. We measured the shear-wave elastic quantitative (SWQ) value of penile CC by 2-D SWE. Erectile function was investigated by intracavernous injection of papaverine, and immunohistochemical (IHC) staining and the western blot analysis to determine the penile CC lesions. After 10 weeks, the SWQ values obtained from penile CC were remarkably higher in the high-cholesterol-fed compared with the control group, and the ΔICP (ICP plateau minus ICP baseline)/MAP (ICP: intracavernous pressure, MAP: mean arterial pressure) was markedly decreased. The IHC staining and western blot revealed extracellular matrix (ECM) accumulation in penile cavernous tissues, and the smooth muscle cell (SMC) phenotypic transition was affected, as indicated by reduced alpha-smooth muscle actin and calponin-1 expression and increased phospho-myosin light chain20 (p-MLC20)/MLC20 and osteopontin expression. Hyperlipidemia resulted in ECM accumulation accompanied with SMC phenotypic transition in penile CC and impaired the erectile function eventually. These might, in turn, lead to variations in the SWQ values. It suggests that 2-D SWE may be a novel, noninvasive and effective approach that distinguishes penile CC lesions secondary to hyperlipidemia from normal.
الموضوعات
Animals , Male , Rabbits , Disease Models, Animal , Elasticity Imaging Techniques/methods , Erectile Dysfunction/etiology , Hyperlipidemias/diagnostic imaging , Penile Erection/physiology , Penis/diagnostic imagingالملخص
Objective To investigate the effect ofdexmedetomidine (Dex) on endoplasmic reticulum stress (ERS) and apoptosis in brain injury after asphyxiating cardiac arrest and cardiopulmonary resuscitation (CA/CPR) in rats.Methods A total of 60 clean male Sprague-Dawley rars were randomly divided into sham-operated group,CA/CPR group and Dex precondition group (n=20).Rats in the control group did not receive CA/CPR;and rats in the CA/CPR group and Dex precondition group were performed cardiac arrest induced by asphyxia,and then,CPR was performed.Dex with dose of 4.0 microgram/kg (body weight) was intravenously injected into rats in the Dex precondition group prior to 5 min of asphyxia.The same volume of saline by intravenous injection was given to rats in control group and CA/CPR group.Brain tissues were collected after the experiment,and wet to dry weight (W/D) ratio was tested.The mRNA expressions of CCAAT-enhancer binding protein homologous protein (CHOP),activation of transcription factor 4 (A TF4) and X-4 box binding protein 1 (XBP1) in the hippocampus were detected by reverse transcription-polymerase chain reaction (RT-PCR).The protein expressions of CHOP,B-cell lymphoma-2 (Bcl-2),Bcl-2 associated X protein (Bax) and cysteinyl aspartate specific proteinase 3 (caspase-3) in the hippocampus were measured by Western boltting.Neuronal apoptosis was detected by terminal dexynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL).Morphological and ultrastructural changes of brains of rats were observed by light microscopy and electron microscopy.Results As compared with the control group,CA/CPR group and Dex precondition group had significantly increased W/D ratio of brain tissues and mRNA expressions of XBP1,A TF4 and CHOP in the hippocampus,significantly higher protein expressions of CHOP,Bax and caspase-3,and statistically lower Bcl-2 expression (P＜0.05).As compared with the CA/CPR group,Dex precondition group had significantly decreased W/D ratio of brain tissues and mRNA expressions of XBP1,A TF4 and CHOP in the hippocampus,significantly lower protein expressions of CHOP,Bax and caspase-3,and statistically higher Bcl-2 expression (P＜0.05).TUNEL indicated that the neuronal apoptosis rate in the control group (7.49％±4.33％) was significantly lower than that in the CA/CPR group and Dex precondition group (29.73％±6.27％ and 16.82％±5.75％,P＜0.05);significant difference was noted between CA/CPR group and Dex precondition group in the neuronal apoptosis rate (P＜0.05).Changes in the morphology and ultramicrostructure injuries of brain tissues were more significant in CA/CPR group,while the changes were obviously alleviated in Dex precondition group.Conclusion Dex can alleviate brain injury after asphyxiating CA/CPR in rats,whose mechanism may be related to ERS and inhibition on apoptosis of nerve cells.
الملخص
Objective To investigate the effect of nerve cells apoptosis induced by endoplasmic reticulum stress (ERS) on brain injury after asphyxiating cardiac arrest and cardiopulmonary resuscitation (CA/CPR) in rats.Methods A total of 40 male Sprague-Dawley rats were randomly divided into control group and CA/CPR group (n=20).The CA/CPR models were established by asphyxia method/CPR.The levels of neuron specific enolization enzyme (NSE) and S100 beta protein (S100β protein) in serum at baseline and 0,3 and 6 h after CPR were detected by double antibody sandwich enzyme-linked immunosorbent assay (ELISA).The mRNA expressions of CCAAT-enhancer binding protein homologous protein (CHOP),activate transcription factor 4 (A TF4) and X-4 box binding protein 1 (XBP1) in the hippocampus were detected by reverse transcription-polymerase chain reaction (RT-PCR).The protein expressions of CHOP,B-cell lymphoma-2 (Bcl-2),Bcl-2 associated X protein (Bax) and cysteinyl aspartate specific proteinase 3 (caspase-3) in the hippocampus were measured by Western boltting.Neuronal apoptosis were detected by terminal dexynucleotidyl transferase (TdT)-mediated dUTP nick end labeling (TUNEL).Morphological and ultrastructural changes of the hippocampi were observed by light microscopy and electron microscopy.Results As compared to that in the control group,S100β protein in the CA/CPR group at 0,3 and 6 h after resuscitation was statistically different (P＜0.05);NSE protein level in the CA/CPR group at 6 h after resuscitation was significantly higher than that in the control group (P＜0.05).As compared with those in the control group,the mRNA expressions of CHOP,A TF4 and XBP-1 in the hippocampus of the CA/CPR group were obviously increased (P＜0.05).Significantly increased protein expressions of CHOP,Bax,and caspase-3,and statistically decreased Bcl-2 expression in the CA/CPR group were noted as compared with those in the control group (P＜0.05).The neuronal apoptosis rate in the CA/CPR group (29.74％±6.26％) was significantly higher than that in the control group (7.48％±4.34％,P＜0.05).The morphology changes and ultramicrostructure injuries of the hippocampus in the CA/CPR group were more obvious as compared with those in the control group.Conclusion CA/CPR in rats causes significant damage to brain tissues,and brain injury is aggravated gradually along with the prolongation of time,and the mechanism of brain injury may be connected with ERS-induced apoptosis of nerve cells.
الملخص
Objective To evaluate the effect of ulinastatin on neuron apoptosis and CCAAT/enhancer-binding protein (CHOP) expression of spinal cord after peripheral nerve injury.Methods A total of 225 healthy male SPF C57BL/6J mice were divided into three groups by using a random number table:sham-operated group,peripheral nerve injury group and ulinastatin group (n=75).The models of unilateral sciatic nerve transection were established in the latter two groups.After the models being established,intraperitoneal injection ofulinastatin 0.2 mL (10 000 U/kg) was performed once daily for 3 consecutive d in ulinastatin group,and the equal volume of normal saline was given once daily for 3 consecutive d in sham-operated group and peripheral nerve injury group.One,3,7,14 and 28 d after surgery,L4-6 spinal cord segments were removed for pathological examination by HE staining,and for detection of neuron apoptosis and apoptotic index (AI) by TUNEL method;the expressions of CHOP,Bcl-2,Bax and cleaved caspase-3 proteins were determined by Western blotting and the ratio of Bcl-2/Bax was calculated,and the CHOP mRNA expression was detected by RT-PCR.Results HE staining showed that the injury of spinal cord in peripheral nerve injury group was more aggravated as compared with that in the sham-operated group,and the injury of spinal cord in ulinastatin group was more alleviated as compared with that in the peripheral nerve injury group.One,3,7,14 and 28 d after surgery,AI was significantly higher,Bcl-2 protein expression was down-regulated,cleaved caspase-3 and Bax protein expressions were up-regulated,Bcl-2/Bax ratio was lower,and CHOP protein or mRNA expressions were up-regulated in the peripheral nerve injury group and ulinastatin group as compared with those in the sham-operated group,with statistically significant differences (P＜0.05).As compared with those in the peripheral nerve injury group,AI was significantly lower,Bcl-2 protein expression was up-regulated,cleaved caspase-3 and Bax protein expressions were down-regulated,Bcl-2/Bax ratio was higher,and CHOP protein or mRNA expressions were down-regulated in ulinastatin group,with statistically significant differences (P＜0.05).Conclusion The mechanism by which ulinastatin protects spinal cord injury after peripheral nerve injury is related to down-regulation of CHOP expression and suppression of neuron apoptosis of spinal cord.
الملخص
Objective To optimize the optimal doses of histamine and 4-aminopyridine (4-AP) in the establish-ment of guinea pigs models of itching, and to establish a new guinea pig model of itching. Methods The central composite design-response surface method was used to arrange the experiment. In the experiment different pruritus agents were hypo-dermically injection of 0. 5 mL in the depilated area, and the scratching incubation period and scratching number in 30 mi-nutes were counted after the injection. The guinea pig itching model was evaluated by observing the behavioral changes of guinea pigs and measuring the levels of histamine and interleukin-6 in the blood. Results The behavioral experiments found that the scratching frequency in the the combination group was significantly higher than the histamine group and 4-AP group (P<0. 01). The itching latency of the combination group was significantly shorter than that of the histamine group and 4-AP group (P<0. 01). Compared with the control group, the histamine concentrations of the combination group and histamine group were significantly increased ( P<0. 05 or P<0. 01 ) , and the level of the combination group was lower than that of the histamine group (P<0. 05). Compared with the control group, the serum IL-6 concentrations of histamine group, 4-AP group and combination group were significantly higher (P<0. 01 or P<0. 05), and those in the combination group were significantly higher than the histamine and 4-AP groups. Compared with the control group, pathologic examina-tion showed proliferation of inflammatory cells in all model groups, and the reaction of the combination group was more ob-vious. Conclusions The optimal conditions used in this experiment are easy to achieve and have good reproducibility in the establishment of a guinea pig model of itching.